TW202525287A - Ionizable lipids - Google Patents

Abstract

The present disclosure provides, among other things, a compound of formula I,

Description

Translated fromChinese

可離子化脂質Ionizable lipids

用於遞送核酸之粒子已成為許多近期工作之主題。某些粒子(例如脂質奈米粒子(LNP))尤其可用於將療法(例如核酸療法)輸送至細胞。參見Tenchov等人，ACS Nano, 2021, 15, 11 16982-17015。LNP尤其包含陽離子或可離子化脂質，其經由靜電相互作用形成囊封核酸且由此促進遞送至細胞中之穩定複合物。Particles for delivering nucleic acids have been the subject of much recent research. Certain particles, such as lipid nanoparticles (LNPs), are particularly useful for delivering therapeutics, such as nucleic acid therapies, to cells.See Tenchovet al. ,ACS Nano , 2021, 15, 11 16982-17015. LNPs, in particular, contain cationic or ionizable lipids that, through electrostatic interactions, form stable complexes that encapsulate nucleic acids and thereby facilitate delivery into cells.

業內仍需要能夠與核酸形成複合物、但相對於先前脂質調配物具有改良之特性(例如，改良之核酸轉染)的陽離子或可離子化脂質。本揭示案提供包含對稱脂族尾之陽離子或可離子化脂質化合物，其令人驚訝地改良包含該等陽離子或可離子化脂質之核酸粒子之特定性質。本揭示案尤其進一步提供製備該等陽離子或可離子化脂質之方法。There remains a need for cationic or ionizable lipids capable of forming complexes with nucleic acids but exhibiting improved properties (e.g., improved nucleic acid transfection) compared to prior lipid formulations. The present disclosure provides cationic or ionizable lipid compounds comprising symmetrical aliphatic tails that surprisingly improve specific properties of nucleic acid particles comprising these cationic or ionizable lipids. Furthermore, the present disclosure provides methods for preparing these cationic or ionizable lipids.

在一些實施例中，本揭示案提供式I化合物：I 或其醫藥學上可接受之鹽，其中 M¹及M²各自為-(CH₂)_p1-； L¹及L²各自選自鍵、-OC(O)-或-C(O)O-； T¹及T²各自為：； L³為視情況經取代之C₁-C₆脂族基； G¹為-OH或-N(R^a)₂，且 R^a為H、視情況經取代之C₁-C₆脂族基或視情況經取代之C₃-C₆環脂族基； p1為選自2-20之整數； p2及p3各自為選自1-20之整數，包括端值，且p2及p3係相同的。In some embodiments, the present disclosure provides compounds of Formula I: I or a pharmaceutically acceptable salt thereof, wherein^M1 and^M2 are each -(_CH2 )_p1- ;^L1 and^L2 are each selected from a bond, -OC(O)- or -C(O)O-;^T1 and^T2 are each:^L3 is an optionally substituted_C1 -_C6 aliphatic group;^G1 is -OH or -N(^Ra )₂ , and^Ra is H, an optionally substituted_C1 -_C6 aliphatic group, or an optionally substituted_C3 -_C6 cycloaliphatic group; p1 is an integer selected from 2 to 20; p2 and p3 are each an integer selected from 1 to 20, inclusive, and p2 and p3 are the same.

在一些實施例中，本揭示案提供一種醫藥組合物，其包含本文所述之化合物(例如陽離子或可離子化脂質，諸如式I化合物)及核酸。In some embodiments, the present disclosure provides a pharmaceutical composition comprising a compound described herein (e.g., a cationic or ionizable lipid, such as a compound of Formula I) and a nucleic acid.

在一些實施例中，本揭示案提供一種懸浮液，其包含分散相及水相，且其中該分散相包含呈粒子形式之如本文所述之醫藥組合物。In some embodiments, the present disclosure provides a suspension comprising a dispersed phase and an aqueous phase, wherein the dispersed phase comprises a pharmaceutical composition as described herein in the form of particles.

在一些實施例中，本揭示案提供一種治療疾病、病症或疾患之方法，其包含向個體投與本文所述之懸浮液。In some embodiments, the present disclosure provides a method of treating a disease, disorder, or condition comprising administering to a subject a suspension described herein.

在一些實施例中，本揭示案提供一種增加個體中之標靶中RNA表現或引起其增加之方法，其包含向個體投與本文所述之懸浮液。In some embodiments, the present disclosure provides a method of increasing or causing an increase in RNA expression in a target in a subject, comprising administering to the subject a suspension described herein.

在一些實施例中，本揭示案提供一種製備式III化合物，III 或其醫藥學上可接受之鹽之方法，該方法包含使式IV-1化合物與式IV-2化合物在熱存在下接觸：以提供第一中間產物，且其中使該第一中間產物與鉑觸媒及第一酸在氫氣存在下接觸以提供第二中間產物，且其中使該第二中間產物與第二酸接觸以提供該式III化合物，其中 M^1’和M^2’獨立地選自鍵或視情況經取代之C₂-C₁₀脂族基； L^1'及L^2'各自獨立地選自鍵、-OC(O)-及-C(O)O-； T^1'及T^2'各自獨立地選自視情況經取代之C₂-C₂₀脂族基； L^3’為視情況經取代之C₁-C₆脂族基； LG¹為適宜之脫離基；且 PG為適宜之醇保護基。In some embodiments, the present disclosure provides a method for preparing a compound of formula III, III or a pharmaceutically acceptable salt thereof, which comprises contacting a compound of formula IV-1 with a compound of formula IV-2 in the presence of heat: to provide a first intermediate product, and wherein the first intermediate product is contacted with a platinum catalyst and a first acid in the presence of hydrogen to provide a second intermediate product, and wherein the second intermediate product is contacted with a second acid to provide the compound of formula III, wherein M^1' and M^2' are independently selected from a bond or an optionally substituted C₂ -C₁₀ aliphatic group; L^{1 '} and L^{2 '} are each independently selected from a bond, -OC(O)-, and -C(O)O-; T^{1 '} and T^{2 '} are each independently selected from an optionally substituted C₂ -C₂₀ aliphatic group; L^3' is an optionally substituted C₁ -C₆ aliphatic group; LG¹ is a suitable leaving group; and PG is a suitable alcohol protecting group.

在一些實施例中，式IV-2化合物藉由以下方式製備：使式V-1a及式V-1b化合物與式V-2化合物在鈀偶合觸媒、銅(I)輔觸媒及胺鹼存在下接觸，以提供該式IV-2化合物，其中LG²為適宜之脫離基。In some embodiments, the compound of formula IV-2 is prepared by reacting the compounds of formula V-1a and formula V-1b with the compound of formula V-2. The reaction is carried out in the presence of a palladium coupling catalyst, a copper (I) cocatalyst and an amine base to provide the compound of formula IV-2, wherein LG² is a suitable detached group.

在一些實施例中，本揭示案提供一種製備式III-1化合物，III-1 或其醫藥學上可接受之鹽之方法，該方法包含使式IV-1化合物與式IV-3化合物在熱存在下接觸：以提供第一中間產物，且其中使該第一中間產物與酸接觸，以提供該式III-1化合物，其中 M^1’和M^2’各自獨立地選自鍵或視情況經取代之C₂-C₁₀脂族基； T^1’及T^2’各自獨立地選自視情況經取代之C₂-C₂₀脂族基； L^3’為視情況經取代之C₁-C₆脂族基； LG¹為適宜之脫離基；且 PG為適宜之醇保護基。In some embodiments, the present disclosure provides a method for preparing a compound of formula III-1, III-1 or a pharmaceutically acceptable salt thereof, the method comprising contacting a compound of formula IV-1 with a compound of formula IV-3 in the presence of heat: to provide a first intermediate product, and wherein the first intermediate product is contacted with an acid to provide the compound of formula III-1, wherein M^1' and M^2' are each independently selected from a bond or an optionally substituted C₂ -C₁₀ aliphatic group; T^1' and T^2' are each independently selected from an optionally substituted C₂ -C₂₀ aliphatic group; L^3' is an optionally substituted C₁ -C₆ aliphatic group; LG¹ is a suitable leaving group; and PG is a suitable alcohol protecting group.

本揭示案尤其提供包含雜環頭基及對稱脂族尾之特定陽離子或可離子化脂質的驚人發現。相對於具有不對稱脂族尾之脂質，此類脂質展現出改良之性質。舉例而言，如本文所提供之實例中所例示，具有雜環頭基及對稱尾之脂質相對於其他脂質調配物展現出改良之轉染。In particular, the present disclosure provides the surprising discovery of specific cationic or ionizable lipids comprising heterocyclic headgroups and symmetric aliphatic tails. These lipids exhibit improved properties relative to lipids with asymmetric aliphatic tails. For example, as exemplified in the examples provided herein, lipids with heterocyclic headgroups and symmetric tails exhibit improved transfection relative to other lipid formulations.

化合物及定義本揭示案之化合物包括上文大體上描述之化合物且藉由本文所揭示之類別、子類及種類進一步說明。如本文使用，除非另有指示，否則下列定義應適用。出於本揭示案之目的，化學元素係根據元素週期表CAS版，Handbook of Chemistry and Physics，第75 版來鑑別。另外，有機化學之一般原理描述於「Organic Chemistry」, Thomas Sorrell, University Science Books, Sausalito: 1999，及「March’s Advanced Organic Chemistry」，第5 版，編輯：Smith, M.B.及March, J., John Wiley & Sons, New York: 2001中，該文獻之全部內容皆以引用方式併入本文中。Compounds and Definitions The compounds of the present disclosure include those generally described above and are further described by the classes, subclasses, and species disclosed herein. As used herein, the following definitions shall apply unless otherwise indicated. For the purposes of this disclosure, chemical elements are identified according to the Periodic Table of the Elements, CAS version, Handbook of Chemistry and Physics, 75th edition. In addition, general principles of organic chemistry are described in "Organic Chemistry", Thomas Sorrell, University Science Books, Sausalito: 1999, and "March's Advanced Organic Chemistry", 5th edition, eds. Smith, MB and March, J., John Wiley & Sons, New York: 2001, the entire contents of which are incorporated herein by reference.

除非另有說明，否則本文所描繪之結構意欲包括該結構之所有立體異構(例如，鏡像異構或非鏡像異構)形式，以及該結構之所有幾何或構形異構形式。舉例而言，預期每一立體中心之R及S構形作為本揭示案之一部分。因此，所提供化合物之單一立體化學異構物以及鏡像異構、非鏡像異構及幾何(或構形)混合物在本揭示案之範圍內。舉例而言，在一些情形下，表1顯示化合物之一或多種立體異構物，且除非另有指示，否則代表單獨及/或呈混合物形式之每一立體異構物。除非另外陳述，否則所提供化合物之所有互變異構形式在本揭示案之範圍內。Unless otherwise indicated, structures depicted herein are intended to include all stereoisomeric (e.g., mirror image or non-mirror image) forms of the structure, as well as all geometric or conformational isomers of the structure. For example, both the R and S configurations of each stereocenter are contemplated as part of this disclosure. Thus, single stereochemical isomers of the provided compounds, as well as mirror image, non-mirror image, and geometric (or conformational) mixtures, are within the scope of this disclosure. For example, in some cases, Table 1 shows one or more stereoisomers of a compound, and unless otherwise indicated, each stereoisomer is represented individually and/or in mixtures. Unless otherwise stated, all tautomeric isomeric forms of the provided compounds are within the scope of this disclosure.

除非另有指示，否則本文所描繪之結構意欲包括不同之處僅在於存在一或多個同位素富集原子的化合物。舉例而言，具有本結構(包括用氘或氚替代氫，或用¹³C或¹⁴C富集碳替代碳)之化合物在本揭示案之範圍內。Unless otherwise indicated, structures depicted herein are intended to include compounds that differ only in the presence of one or more isotopically enriched atoms. For example, compounds having the present structures including the replacement of a hydrogen by a deuterium or tritium, or the replacement of a carbon by a¹³ C- or¹⁴ C-enriched carbon are within the scope of this disclosure.

約或大約：如本文所用之術語「大約」或「約」在應用於一或多個所關注值時係指與所述參考值相似之值。一般而言，熟悉上下文之熟習此項技術者應瞭解該上下文中由「約」或「大約」涵蓋之相關變化程度。舉例而言，在一些實施例中，術語「大約」或「約」可涵蓋在所提及值之(亦即±) 25%、20%、19%、18%、17%、16%、15%、14%、13%、12%、11%、10%、9%、8%、7%、6%、5%、4%、3%、2%、1%或更小內之一系列值。About or approximately: As used herein, the terms "about" or "approximately" when applied to one or more referenced values refer to values that are similar to the reference value. Generally, one skilled in the art familiar with the context will understand the relevant degree of variation encompassed by "about" or "approximately" in that context. For example, in some embodiments, the terms "about" or "approximately" can encompass a range of values that are within (i.e., ±) 25%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1% or less of the referenced value.

投與：如本文所用之術語「投與(administering)」或「投與(administration)」通常係指向個體投與組合物以達成為組合物或包括在組合物中之劑至靶位點或欲治療位點之遞送。熟習此項技術者應意識到在適當情況下可用於投與個體(例如人類)之多種途徑。舉例而言，在一些實施例中，投與可為眼部、口服、非經腸、局部等。在一些具體實施例中，投與可為支氣管(例如，藉由支氣管滴注)、頰側、真皮(其可為或包括例如真皮局部、真皮內、真皮間、穿真皮等中之一或多者)、腸、動脈內、真皮內、胃內、髓內、肌內、鼻內、腹膜內、鞘內、靜脈內、室內、特定器官內(例如，肝內)、黏膜、鼻、口服、直腸、皮下、舌下、局部、氣管(例如，藉由氣管內滴注)、陰道、玻璃體等。在一些實施例中，投與可為非經腸。在一些實施例中，投與可為口服。在一些具體實施例中，投與可為靜脈內。在一些具體實施例中，投與可為皮下。在一些實施例中，投與可僅涉及單一劑量。在一些實施例中，投與可涉及施用固定數量之劑量。在一些實施例中，投與可涉及間歇給藥(例如，在時間上分開之複數個劑量)及/或週期性(例如，由共同時間段分開之個別劑量)給藥。在一些實施例中，投與可涉及持續至少所選時間段之連續給藥(例如，灌注)。在一些實施例中，投與可包括初免及加強方案(prime-and-boost protocol)。初免及加強方案可包括投與第一劑量之醫藥組合物(例如免疫原性組合物，例如疫苗)，然後在時間間隔後，投與第二或後續劑量之醫藥組合物(例如免疫原性組合物，例如疫苗)。在免疫原性組合物之情形下，初免及加強方案可導致患者中之免疫反應增加。Administration: As used herein, the terms "administering" or "administration" generally refer to administering a composition to a subject to achieve delivery of the composition or an agent included in the composition to a target site or site of treatment. Those skilled in the art will recognize that a variety of routes of administration can be used to administer a composition to a subject (e.g., a human), as appropriate. For example, in some embodiments, administration can be ocular, oral, parenteral, topical,or the like. In some embodiments, administration can be bronchial (e.g. , by bronchial instillation), buccal, dermal (which can be or include, for example, one or more of topical, intradermal, interdermal, transdermal,etc. ), intestinal, intraarterial, intradermal, intragastric, intramedullary, intramuscular, intranasal, intraperitoneal, intrathecal, intravenous, intraventricular, intraorgan (e.g. , intrahepatic), mucosal, nasal, oral, rectal, subcutaneous, sublingual, topical, tracheal (e.g. , by intratracheal instillation), vaginal, vitreous,etc. In some embodiments, administration can be parenteral. In some embodiments, administration can be oral. In some embodiments, administration can be intravenous. In some embodiments, administration can be subcutaneous. In some embodiments, administration can involve only a single dose. In some embodiments, administration may involve administering a fixed amount of a dose. In some embodiments, administration may involve intermittent dosing (e.g. , multiple doses separated in time) and/or periodic dosing (e.g. , individual doses separated by a common time period). In some embodiments, administration may involve continuous dosing (e.g. , perfusion) that lasts for at least a selected time period. In some embodiments, administration may include a prime-and-boost protocol. A prime-and-boost protocol may include administering a first dose of a pharmaceutical composition (e.g., an immunogenic composition, such as a vaccine), followed by administering a second or subsequent dose of a pharmaceutical composition (e.g., an immunogenic composition, such as a vaccine) after a time interval. In the case of an immunogenic composition, a prime-and-boost protocol may result in an increased immune response in the patient.

脂族：術語「脂族」係指完全飽和或含有一或多個不飽和單元之直鏈(亦即，無分枝)或分枝、經取代或未經取代之烴鏈，或完全飽和或含有一或多個不飽和單元、但不為芳族、與分子之其餘部分具有單一連接點或一個以上之連接點的單環烴或二環烴(在本文中亦稱為「環脂族」)。除非另外指定，否則脂族基團含有1-12個脂族碳原子。在一些實施例中，脂族基團含有1-6個脂族碳原子(例如，C_1-6)。在一些實施例中，脂族基團含有1-5個脂族碳原子(例如，C_1-5)。在其他實施例中，脂族基團含有1-4個脂族碳原子(例如，C_1-4)。在其他實施例中，脂族基團含有1-3個脂族碳原子(例如，C_1-3)，且在其他實施例中，脂族基團含有1-2個脂族碳原子(例如，C_1-2)。適宜脂族基團包括(但不限於)直鏈或分枝、經取代或未經取代之烷基、烯基或炔基及其雜合物。較佳脂族基團係C_1-6烷基。Aliphatic: The term "aliphatic" refers to a straight (i.e., unbranched) or branched, substituted or unsubstituted hydrocarbon chain that is fully saturated or contains one or more unsaturated units, or a monocyclic or bicyclic hydrocarbon that is fully saturated or contains one or more unsaturated units but is not aromatic and has a single point of attachment or more than one point of attachment to the rest of the molecule (also referred to herein as a "cycloaliphatic"). Unless otherwise specified, an aliphatic group contains 1-12 aliphatic carbon atoms. In some embodiments, an aliphatic group contains 1-6 aliphatic carbon atoms (e.g.,_C1-6 ). In some embodiments, an aliphatic group contains 1-5 aliphatic carbon atoms (e.g.,_C1-5 ). In other embodiments, the aliphatic group contains 1-4 aliphatic carbon atoms (e.g., C_1-4 ). In other embodiments, the aliphatic group contains 1-3 aliphatic carbon atoms (e.g., C_1-3 ), and in other embodiments, the aliphatic group contains 1-2 aliphatic carbon atoms (e.g., C_1-2 ). Suitable aliphatic groups include, but are not limited to, linear or branched, substituted or unsubstituted alkyl, alkenyl, or alkynyl groups, and hybrids thereof. Preferred aliphatic groups are C_1-6 alkyl groups.

烷基：單獨使用或作為較大部分之一部分使用之術語「烷基」係指具有(除非另外指定) 1-12個、1-10個、1-8個、1-6個、1-4個、1-3個或1-2個碳原子之飽和、視情況經取代之直鏈或分枝鏈烴基(例如，C_1-12、C_1-10、C_1-8、C_1-6、C_1-4、C_1-3或C_1-2)。示範性烷基包括甲基、乙基、丙基、丁基、戊基、己基及庚基。Alkyl: The term "alkyl," used alone or as part of a larger group, refers to a saturated, optionally substituted, straight or branched chain alkyl group (e.g.,_C1-12 , C1-10, C1-8, C1-6, C1-4, C1-3, or C1-2) having, unless otherwise specified, 1-12,_1-10 ,_1-8 ,_1-6 ,_1-4 ,_1-3 , or_1-2 carbon atoms. Exemplary alkyl groups include methyl, ethyl, propyl, butyl, pentyl, hexyl, and heptyl.

伸烷基：術語「伸烷基」係指二價烷基。在一些實施例中，「伸烷基」係二價直鏈或分枝烷基。在一些實施例中，「伸烷基鏈」係聚亞甲基，亦即-(CH₂)_n-，其中n係例如1至6、1至4、1至3、1至2、或2至3之正整數。視情況經取代之伸烷基鏈係其中一或多個亞甲基氫原子視情況經取代基替代之聚亞甲基。適宜取代基包括下文針對經取代脂族基團描述之取代基且亦包括本文說明書中所述之取代基。應瞭解，伸烷基之兩個取代基可一起形成環系統。在某些實施例中，兩個取代基可一起形成3至7員環。取代基可處於相同或不同的原子上。後綴「-伸基(-ene)」或「-伸基(-enyl)」在附加至本文之某些基團時欲指該基團之雙官能部分。舉例而言，「-伸基(-ene)」或「-伸基(-enyl)」在附加至「環丙基」時變成「伸環丙基(cyclopropylene)」或「伸環丙基(cyclopropylenyl)」且欲指雙官能環丙基，例如。Alkylene: The term "alkylene" refers to a divalent alkyl group. In some embodiments, an "alkylene" is a divalent linear or branched alkyl group. In some embodiments, an "alkylene chain" is a polymethylene group, i.e., -(_CH2 )_n- , where n is a positive integer, for example, 1 to 6, 1 to 4, 1 to 3, 1 to 2, or 2 to 3. An optionally substituted alkylene chain is a polymethylene group in which one or more methylene hydrogen atoms are optionally replaced with a substituent. Suitable substituents include those described below for substituted aliphatic groups and also include those described herein. It should be understood that two substituents of an alkylene group may be taken together to form a ring system. In certain embodiments, two substituents may be taken together to form a 3- to 7-membered ring. The substituents may be on the same or different atoms. The suffix "-ene" or "-enyl" when appended to certain groups herein is intended to refer to the difunctional portion of the group. For example, "-ene" or "-enyl" when appended to "cyclopropyl" becomes "cyclopropylene" or "cyclopropylenyl" and is intended to refer to the difunctional cyclopropyl group, e.g. .

烯基：單獨使用或作為較大部分之一部分使用之術語「烯基」係指具有至少一個雙鍵且具有(除非另外指定) 2-12個、2-10個、2-8個、2-6個、2-4個或2-3個碳原子之視情況經取代之直鏈或分枝鏈或環狀烴基(例如，C_2-12、C_2-10、C_2-8、C_2-6、C_2-4或C_2-3)。示範性烯基包括乙烯基、丙烯基、丁烯基、戊烯基、己烯基及庚烯基。術語「環烯基」係指含有至少一個碳-碳雙鍵且具有約3至約10個碳原子之視情況經取代之非芳族單環或多環系統。示範性單環環烯基環包括環戊烯基、環己烯基及環庚烯基。Alkenyl: The term "alkenyl," used alone or as part of a larger group, refers to an optionally substituted, straight or branched chain or cyclic alkyl group (e.g., C2-12, C2-10, C2-8, C2-6, C2-4, or_{C2-3) having at least one double bond and, unless otherwise specified, 2-12, 2-10, 2-8, 2-6, 2-4, or 2-3carbonatoms.Exemplaryalkenyl} groups include ethenyl, propenyl, butenyl, pentenyl, hexenyl, and heptenyl. The term "cycloalkenyl" refers to an optionally substituted, non-aromatic, monocyclic or polycyclic ring system containing at least one carbon-carbon double bond and having from about 3 to about 10 carbon atoms. Exemplary monocyclic cycloalkenyl rings include cyclopentenyl, cyclohexenyl, and cycloheptenyl.

炔基：單獨使用或作為較大部分之一部分使用之術語「炔基」係指具有至少一個三鍵且具有(除非另外指定) 2-12個、2-10個、2-8個、2-6個、2-4個或2-3個碳原子之視情況經取代之直鏈或分枝鏈烴基(例如，C_2-12、C_2-10、C_2-8、C_2-6、C_2-4或C_2-3)。示範性炔基包括乙炔基、丙炔基、丁炔基、戊炔基、己炔基及庚炔基。Alkynyl: The term "alkynyl," used alone or as part of a larger group, refers to an optionally substituted straight or branched chain alkyl group (e.g., C2-12, C2-10, C2-8, C2-6, C2-4, or C2-3) having at least one triple bond and, unless otherwise specified, 2-12,_2-10 ,_{2-8, 2-6, 2-4,or2-3} carbon atoms. Exemplary alkynyl groups include ethynyl, propynyl, butynyl, pentynyl, hexynyl, and heptynyl.

芳基：術語「芳基」係指具有總共六至十四個環成員之單環及二環系統(例如，C₆-C₁₄)，其中系統中之至少一個環係芳族且其中系統中之每一環含有三至七個環成員。在一些實施例中，「芳基」含有六至十二個總環成員(例如，C₆-C₁₂)。術語「芳基」可與術語「芳基環」互換使用。在某些實施例中，「芳基」係指可帶有一或多個取代基之芳族環系統，包括但不限於苯基、聯苯基、萘基、蒽基及其類似基團。除非另外指定，否則「芳基」係烴。在一些實施例中，「芳基」環系統係稠合至非芳族環(例如，環烷基)之芳族環(例如，苯基)。稠合芳基環之實例包括、及。Aryl: The term "aryl" refers to monocyclic and bicyclic ring systems having six to fourteen total ring members (e.g.,_C6 -_C14 ), wherein at least one ring in the system is aromatic and wherein each ring in the system contains three to seven ring members. In some embodiments, "aryl" contains six to twelve total ring members (e.g.,_C6 -_C12 ). The term "aryl" is used interchangeably with the term "aryl ring." In certain embodiments, "aryl" refers to an aromatic ring system that may bear one or more substituents, including but not limited to phenyl, biphenyl, naphthyl, anthracenyl, and the like. Unless otherwise specified, "aryl" is hydrocarbon. In some embodiments, an "aryl" ring system is an aromatic ring (e.g., phenyl) fused to a non-aromatic ring (e.g., cycloalkyl). Examples of fused aryl rings include 、 and .

生物樣品：如本文所用之術語「生物樣品」通常係指自所關注生物來源(例如，組織或生物體或細胞培養物)獲得或衍生而來之樣品，如本文所述。在一些實施例中，所關注來源包含生物體，例如動物或人類。在一些實施例中，生物樣品係或包含生物組織或流體。在一些實施例中，生物樣品可為或包含骨髓；血液；血球；腹水；組織或細針生檢樣品；含細胞體液；自由漂浮核酸；痰；唾液；尿；腦脊液、腹膜液；肋膜液；糞便；淋巴；婦科液；皮膚拭子；陰道拭子；口腔拭子；鼻拭子；洗滌液或灌洗液，例如導管灌洗液或支氣管肺泡灌洗液；抽吸物；刮擦物；骨髓樣本；組織生檢樣本；手術樣本；糞便、其他體液、分泌物及/或排洩物；及/或來自其之細胞等。在一些實施例中，生物樣品係或包含自個體獲得之細胞。在一些實施例中，所獲得細胞係或包括自其獲得樣品之個體之細胞。在一些實施例中，樣品係藉由任何適當方式自所關注來源直接獲得之「初級樣品」。舉例而言，在一些實施例中，初級生物樣品係藉由選自由生檢(例如，細針抽吸或組織生檢)、手術、收集體液(例如，血液、淋巴、糞便等)等組成之群之方法獲得。在一些實施例中，如根據上下文應清楚，術語「樣品」係指藉由處理初級樣品(例如，藉由去除初級樣品之一或多種組分及/或藉由將一或多種劑添加至初級樣品中)獲得之製劑。舉例而言，使用半透膜過濾。此「經處理樣品」可包含例如自樣品提取或藉由使初級樣品經歷諸如擴增或反轉錄mRNA、分離及/或純化某些組分等技術獲得之核酸或蛋白質。Biological sample: As used herein, the term "biological sample" generally refers to a sample obtained or derived from a biological source of interest (e.g., a tissue, an organism, or a cell culture), as described herein. In some embodiments, the source of interest comprises an organism, such as an animal or a human. In some embodiments, the biological sample comprises or includes a biological tissue or fluid. In some embodiments, a biological sample can be or comprise bone marrow; blood; blood cells; ascites; tissue or fine needle biopsy samples; cell-containing body fluids; free-floating nucleic acids; sputum; saliva; urine; cerebrospinal fluid, peritoneal fluid; pleural fluid; feces; lymph; gynecological fluid; skin swab; vaginal swab; oral swab; nasal swab; washes or lavages, such as catheter washes or bronchoalveolar lavage; aspirates; scrapings; bone marrow samples; tissue biopsy samples; surgical samples; feces, other body fluids, secretions, and/or excretions; and/or cells therefrom. In some embodiments, a biological sample is or comprises cells obtained from an individual. In some embodiments, the cells obtained are or include cells from the individual from which the sample was obtained. In some embodiments, the sample is a "primary sample" obtained directly from a source of interest by any appropriate means. For example, in some embodiments, the primary biological sample is obtained by a method selected from the group consisting of biopsy (e.g., fine needle aspiration or tissue biopsy), surgery, collection of body fluids (e.g., blood, lymph, feces, etc.). In some embodiments, as should be clear from the context, the term "sample" refers to a preparation obtained by processing the primary sample (e.g., by removing one or more components of the primary sample and/or by adding one or more agents to the primary sample). For example, filtration using a semipermeable membrane. Such a "processed sample" may include, for example, nucleic acids or proteins extracted from the sample or obtained by subjecting the primary sample to techniques such as amplification or reverse transcription of mRNA, isolation and/or purification of certain components.

碳環基：如本文所用之術語「碳環基」、「碳環(carbocycle)」及「碳環(carbocyclic ring)」係指如本文所述具有3至14個成員之飽和或部分不飽和環狀脂族單環、雙環或多環系統，其中該脂族環系統視情況如本文所述經取代。碳環基團包括但不限於環丙基、環丁基、環戊基、環戊烯基、環己基、環己烯基、環庚基、環庚烯基、環辛基、環辛烯基、降莰基、金剛烷基及環辛二烯基。在一些實施例中，「碳環基」(或「環脂族」)係指視情況經取代之單環C₃-C₈烴，或視情況經取代之C₇-C₁₀雙環烴，其完全飽和或含有一或多個不飽和單元，但不是芳族的，與分子的其餘部分有一個連接點。術語「環烷基」係指約3至約10個環碳原子之視情況經取代之飽和環系統。在一些實施例中，環烷基具有3–6個碳。示範性單環環烷基環包括環丙基、環丁基、環戊基、環己基及環庚基。術語「環烯基」係指含有至少一個碳-碳雙鍵且具有約3至約10個碳原子之視情況經取代之非芳族單環或多環系統。示範性單環環烯基環包括環戊烯基、環己烯基及環庚烯基。Carbocyclyl: As used herein, the terms "carbocyclyl,""carbocycle," and "carbocyclic ring" refer to a saturated or partially unsaturated cyclic aliphatic monocyclic, bicyclic, or polycyclic ring system having from 3 to 14 members, as described herein, wherein the aliphatic ring system is optionally substituted as described herein. Carbocyclyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cycloheptyl, cycloheptenyl, cyclooctyl, cyclooctenyl, norbornyl, adamantyl, and cyclooctadienyl. In some embodiments, "carbocyclyl" (or "cycloaliphatic") refers to an optionally substituted monocyclic_C3 -_C8 hydrocarbon, or an optionally substituted_C7 -_C10 bicyclic hydrocarbon, which is fully saturated or contains one or more unsaturated units, but is not aromatic, and has one point of attachment to the rest of the molecule. The term "cycloalkyl" refers to an optionally substituted saturated ring system of about 3 to about 10 ring carbon atoms. In some embodiments, the cycloalkyl group has 3-6 carbon atoms. Exemplary monocyclic cycloalkyl rings include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cycloheptyl. The term "cycloalkenyl" refers to an optionally substituted non-aromatic monocyclic or polycyclic ring system containing at least one carbon-carbon double bond and having from about 3 to about 10 carbon atoms. Exemplary monocyclic cycloalkenyl rings include cyclopentenyl, cyclohexenyl, and cycloheptenyl.

載劑：如本文所用之術語「載劑」係指與組合物一起投與之稀釋劑、佐劑、賦形劑或媒劑。在一些示範性實施例中，載劑可包括無菌液體，例如水及油，包括石油、動物油、植物油或合成來源之油，例如花生油、大豆油、礦物油、芝麻油及諸如此類。在一些實施例中，載劑係或包括一或多種固體組分。Carrier: As used herein, the term "carrier" refers to a diluent, adjuvant, excipient, or vehicle with which a composition is administered. In some exemplary embodiments, a carrier may include sterile liquids such as water and oils, including those of petroleum, animal, vegetable, or synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil, and the like. In some embodiments, a carrier is or includes one or more solid components.

組合療法：如本文所用之術語「組合療法」係指其中將個體同時暴露於兩種或更多種治療方案(例如，兩種或更多種治療劑或模式)之彼等情況。在一些實施例中，兩種或更多種方案可同時投與；在一些實施例中，該等方案可依序投與(例如，在投與第二方案之任何劑量之前投與第一方案之所有「劑量」)；在一些實施例中，該等劑係以重疊給藥方案投與。在一些實施例中，組合療法之「投與」可涉及向接受組合中之一或多種劑或模式之個體投與另一(些)劑或模式。為簡潔起見，組合療法不要求在單一組合物中一起投與個別劑(或甚至必要地同時投與)，但在一些實施例中，兩種或更多種劑或其活性部分可在組合組合物中或甚至在組合化合物中一起投與(例如，作為單一化學複合物或共價實體之一部分)。Combination therapy: As used herein, the term "combination therapy" refers to situations in which a subject is exposed to two or more treatment regimens (e.g., two or more therapeutic agents or modalities) simultaneously. In some embodiments, the two or more regimens are administered simultaneously; in some embodiments, the regimens are administered sequentially (e.g., all "doses" of the first regimen are administered before any doses of the second regimen are administered); in some embodiments, the agents are administered in an overlapping dosing schedule. In some embodiments, "administration" of a combination therapy may involve administering to a subject receiving one or more agents or modalities of the combination the other(s). For the sake of brevity, combination therapy does not require that the individual agents be administered together in a single composition (or even necessarily simultaneously), but in some embodiments, two or more agents, or active portions thereof, can be administered together in a combination composition or even in a combination compound (e.g., as part of a single chemical complex or covalent entity).

相當：如本文所用之術語「相當」係指可能彼此不一致、但足夠相似以容許在其之間進行比較之兩種或更多種劑、實體、情況、條件集等，以使得熟習此項技術者應意識到，可基於所觀察到之差異或相似性合理地得出結論。在一些實施例中，相當之條件集、情況、個體或群體之特徵在於複數個實質上一致之特徵及一個或少量不同之特徵。在上下文中，熟習此項技術者將理解，在任何給定環境下需要何種程度之一致性才能使二或更多種此類劑、實體、情況、條件集合等被視為可比較的。舉例而言，熟習此項技術者應意識到，當特徵在於足夠數量及類型之實質上一致之特徵時，情況集、個體或群體係彼此相當的以保證合理的結論，即在不同的情況集、個體或群體下或使用不同的情況集、個體或群體獲得之結果或觀測到之現象的差異係由彼等不同特徵之變化導致或指示該變化。Equivalent : As used herein, the term "equivalent" refers to two or more agents, entities, situations, sets of conditions, etc. that may not be identical to one another, but are sufficiently similar to permit comparison between them, such that one skilled in the art would recognize that conclusions can reasonably be drawn based on the observed differences or similarities. In some embodiments, equivalent sets of conditions, situations, individuals, or groups are characterized by a plurality of substantially identical characteristics and one or a small number of different characteristics. In this context, one skilled in the art will understand what degree of identity is required in any given set of circumstances for two or more such agents, entities, situations, sets of conditions, etc. to be considered comparable. For example, one skilled in the art will recognize that sets of situations, individuals, or groups are equivalent to one another when characterized by a sufficient number and type of substantially identical features to warrant a reasonable conclusion that differences in results or observed phenomena obtained under or using different sets of situations, individuals, or groups are caused by or indicative of variations in those different features.

組合物：熟習此項技術者應意識到，術語「組合物」可用於指包含一或多種指定組分之離散物理實體。一般而言，除非另外指定，否則組合物可具有任一形式，例如氣體、凝膠、液體、固體等。Composition: Those skilled in the art will appreciate that the term "composition" may be used to refer to a discrete physical entity comprising one or more specified components. Generally, unless otherwise specified, a composition may have any form, such as gas, gel, liquid, solid, etc.

環脂族：如本文所用之術語「環脂族」係指完全飽和或含有一或多個不飽和單元、但不為芳族、與分子之其餘部分具有單一連接點或一個以上之連接點的單環C_3-8烴或二環C_6-10烴。Cycloaliphatic : As used herein, the term "cycloaliphatic" refers to a monocyclic C3-8 hydrocarbon or a bicyclic C6-10 hydrocarbon that is fully saturated or contains one or more unsaturated units, but is not aromatic, and has a single point of attachment or more than one_point of attachment to the_rest of the molecule.

環烷基：如本文所用之術語「環烷基」係指約3至約10個環碳原子之視情況經取代之飽和單環或多環系統。示範性單環環烷基環包括環丙基、環丁基、環戊基、環己基及環庚基。Cycloalkyl : As used herein, the term "cycloalkyl" refers to an optionally substituted, saturated, monocyclic or polycyclic ring system of about 3 to about 10 ring carbon atoms. Exemplary monocyclic cycloalkyl rings include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cycloheptyl.

去氧核糖核酸(DNA)：如本文所用之術語「DNA」係指通常為雙股且包含腺嘌呤、胞嘧啶、鳥嘌呤及胸腺嘧啶之核苷酸之聚合物分子，及如定義「核酸/多核苷酸」中所指定之去氧核糖主鏈結構。在一些實施例中，DNA係線性DNA、質體DNA、微環DNA、奈米質體DNA、狗骨DNA或轉座子。Deoxyribonucleic acid (DNA) : As used herein, the term "DNA" refers to a polymeric molecule that is typically double-stranded and comprises the nucleotides adenine, cytosine, guanine, and thymine, and a deoxyribose backbone structure as specified in the definition of "nucleic acid/polynucleotide." In some embodiments, the DNA is linear DNA, plasmid DNA, minicircle DNA, nanoplasmid DNA, dogbone DNA, or a transposon.

去氧核糖核苷酸：如本文所用之術語「去氧核糖核苷酸」係指未經修飾及經修飾之去氧核糖核苷酸。舉例而言，未經修飾之去氧核糖核苷酸包括嘌呤鹼基腺嘌呤(A)及鳥嘌呤(G)，以及嘧啶鹼基胞嘧啶(C)及胸腺嘧啶(T)。經修飾之去氧核糖核苷酸可包括一或多種修飾，包括但不限於例如(a)末端修飾，例如5'末端修飾(例如，磷酸化、去磷酸化、偶聯、反向鍵聯等)、3'末端修飾(例如，偶聯、反向鍵聯等)，(b)鹼基修飾，例如用經修飾鹼基、穩定鹼基、去穩定鹼基或與擴展的配偶體譜系鹼基配對之鹼基或共軛鹼基替代，(c)糖修飾(例如，在2'位或4'位)或替代糖，及(d)核苷間鍵聯修飾，包括修飾或替代磷酸二酯鍵聯。Deoxyribonucleotides: As used herein, the term "deoxyribonucleotides" refers to both unmodified and modified deoxyribonucleotides. For example, unmodified deoxyribonucleotides include the purine bases adenine (A) and guanine (G), and the pyrimidine bases cytosine (C) and thymine (T). Modified deoxyribonucleotides can include one or more modifications, including but not limited to, for example, (a) terminal modifications,such as 5' terminal modifications (e.g. , phosphorylation, dephosphorylation, conjugation, reverse linkage, etc. ), 3' terminal modifications (e.g. , conjugation, reverse linkage, etc. ), (b) base modifications,such as replacement with a modified base, a stable base, a destabilized base, or a base that pairs with an expanded spectrum of partner bases or a conjugated base, (c) sugar modifications (e.g. , at the 2' or 4' position) or replacement sugars, and (d) internucleoside linkage modifications, including modification or replacement of phosphodiester linkages.

給藥方案或治療方案：熟習此項技術者應意識到，術語「給藥方案」及「治療方案」可用於指個別地投與個體之一組單位劑量(通常一個以上)，其通常由時間段分開。在一些實施例中，給定治療劑具有推薦的給藥方案，其可涉及一或多個劑量。在一些實施例中，給藥方案包括複數個劑量，每一劑量在時間上與其他劑量分開。在一些實施例中，個別劑量由相同長度之時間段彼此分開；在一些實施例中，給藥方案包括複數個劑量及分開個別劑量之至少兩個不同時間段。在一些實施例中，給藥方案內之所有劑量具有相同的單位劑量量。在一些實施例中，給藥方案內之不同劑量具有不同量。在一些實施例中，給藥方案包括第一劑量量之第一劑量，然後係不同於第一劑量量之第二劑量量之一或多個額外劑量。在一些實施例中，給藥方案包括第一劑量量之第一劑量，然後係與第一劑量量相同之第二劑量量之一或多個額外劑量。在一些實施例中，當投與相關群體時，給藥方案與期望或有益結果相關聯(亦即，係治療給藥方案)。Dosing regimen ortreatment regimen: Those skilled in the art will appreciate that the terms "dosing regimen" and "treatment regimen" may be used to refer to a set of unit doses (usually more than one) that are individually administered to a subject, typically separated by time periods. In some embodiments, a given treatment has a recommended dosing regimen that may involve one or more doses. In some embodiments, the dosing regimen includes a plurality of doses, each dose separated from the other doses in time. In some embodiments, the individual doses are separated from each other by time periods of the same length; in some embodiments, the dosing regimen includes a plurality of doses and at least two different time periods separating the individual doses. In some embodiments, all doses within a dosing regimen have the same unit dose amount. In some embodiments, different doses within a dosing regimen have different amounts. In some embodiments, a dosing regimen includes a first dose of a first dose amount, followed by one or more additional doses of a second dose amount that is different from the first dose amount. In some embodiments, a dosing regimen includes a first dose of a first dose amount, followed by one or more additional doses of a second dose amount that is the same as the first dose amount. In some embodiments, a dosing regimen is associated with a desired or beneficial outcome when administered to a population of interest (i.e., a therapeutic dosing regimen).

賦形劑：如本文所用之術語「賦形劑」係指可納入醫藥組合物中例如以提供或有助於期望一致性或穩定效應之非治療劑。適宜醫藥賦形劑包括例如澱粉、葡萄糖、乳糖、蔗糖、明膠、麥芽、水稻、麵粉、白堊、矽膠、硬脂酸鈉、單硬脂酸甘油酯、滑石、氯化鈉、脫脂奶粉、甘油、丙二醇、水、乙醇及諸如此類。Excipients: As used herein, the term "excipient" refers to non-therapeutic agents that can be included in a pharmaceutical composition, for example, to provide or contribute to a desired consistency or stability. Suitable pharmaceutical excipients include, for example, starch, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glyceryl monostearate, talc, sodium chloride, skimmed milk powder, glycerin, propylene glycol, water, ethanol, and the like.

雜脂族：如本文所用之術語「雜脂族」或「雜脂族基團」表示除碳原子外具有一至五個雜原子之視情況經取代之烴部分，其可為直鏈(亦即，無分枝)、分枝或環狀(「雜環」)且可為完全飽和的或可含有一或多個不飽和單元，但不為芳族。術語「雜原子」係指氮、氧或硫，且包括氮或硫之任何氧化形式，及鹼性氮之任何四級銨化形式。術語「氮」亦包括經取代之氮。除非另外指定，否則雜脂族基團含有1-10個碳原子，其中1-3個碳原子視情況地且獨立地經選自氧、氮及硫之雜原子替代。在一些實施例中，雜脂族基團含有1-4個碳原子，其中1-2個碳原子視情況地且獨立地經選自氧、氮及硫之雜原子替代。在其他實施例中，雜脂族基團含有1-3個碳原子，其中1個碳原子視情況地且獨立地經選自氧、氮及硫之雜原子替代。適宜雜脂族基團包括(但不限於)直鏈或分枝雜烷基、雜烯基及雜炔基。舉例而言，1至10個原子之雜脂族基團包括以下示範性基團：-O-CH₃、-CH₂-O-CH₃、-O-CH₂-CH₂-O-CH₂-CH₂-O-CH₃及諸如此類。Heteroaliphatic : As used herein, the term "heteroaliphatic" or "heteroaliphatic group" refers to an optionally substituted hydrocarbon moiety having one to five heteroatoms in addition to carbon atoms, which may be linear (i.e., unbranched), branched, or cyclic ("heterocyclic") and may be fully saturated or may contain one or more unsaturated units, but is not aromatic. The term "heteroatom" refers to nitrogen, oxygen, or sulfur, and includes any oxidized form of nitrogen or sulfur, and any quaternary ammonium form of basic nitrogen. The term "nitrogen" also includes substituted nitrogen. Unless otherwise specified, a heteroaliphatic group contains 1-10 carbon atoms, of which 1-3 carbon atoms are optionally and independently replaced by heteroatoms selected from oxygen, nitrogen, and sulfur. In some embodiments, the heteroaliphatic group contains 1-4 carbon atoms, wherein 1-2 carbon atoms are optionally and independently replaced by a heteroatom selected from oxygen, nitrogen, and sulfur. In other embodiments, the heteroaliphatic group contains 1-3 carbon atoms, wherein 1 carbon atom is optionally and independently replaced by a heteroatom selected from oxygen, nitrogen, and sulfur. Suitable heteroaliphatic groups include, but are not limited to, straight-chain or branched heteroalkyl groups, heteroalkenyl groups, and heteroalkynyl groups. For example, heteroaliphatic groups of 1 to 10 atoms include the following exemplary groups: -O-_CH3 , -CH2-O-_CH3 , -O-_CH2 -_CH2 -_O -CH2-_CH2 -O-_CH3, and the like.

雜芳基：單獨使用或作為較大部分(例如，「雜芳烷基」或「雜芳烷氧基」)之一部分使用之術語「雜芳基」及「雜芳-」係指具有5至10個環原子(例如，5至6員單環雜芳基或9至10員雙環雜芳基)；具有在環狀陣列中共享之6、10或14個π電子；及除碳原子外具有一至五個雜原子之單環或雙環基團。雜芳基包括但不限於噻吩基、呋喃基、吡咯基、咪唑基、吡唑基、三唑基、四唑基、噁唑基、異噁唑基、噁二唑基、噻唑基、異噻唑基、噻二唑基、吡啶基、噠嗪基、嘧啶基、吡嗪基、吲嗪基、嘌呤基、萘啶基、蝶啶基、咪唑并[1,2-a]嘧啶基、咪唑并[1,2-a]吡啶基、咪唑并[4,5-b]吡啶基、咪唑并[4,5-c]吡啶基、吡咯并吡啶基、吡咯并吡嗪基、噻吩并嘧啶基、三唑并吡啶基及苯并異噁唑基。如本文所用，術語「雜芳基」及「雜芳-」亦包括如下基團，其中雜芳族環融合至一或多個芳基、環脂族或雜環基環，其中連接基團或連接點係在雜芳族環(亦即，具有1至3個雜原子之雙環雜芳基環)上。非限制性實例包括吲哚基、異吲哚基、苯并噻吩基、苯并呋喃基、二苯并呋喃基、吲唑基、苯并咪唑基、苯并三唑基、苯并噻唑基、苯并噻二唑基、苯并噁唑基、喹啉基、異喹啉基、㖕啉基、呔嗪基、喹唑啉基、喹喔啉基、4H-喹嗪基、咔唑基、吖啶基、啡嗪基、啡噻嗪基、啡噁嗪基、四氫喹啉基、四氫異喹啉基、吡啶并[2,3-b]-1,4-噁嗪-3(4H)-酮、4H-噻吩并[3,2-b]吡咯及苯并異噁唑基。術語「雜芳基」可與術語「雜芳基環」、「雜芳基」或「雜芳族」互換使用，該等術語中之任一者包括視情況經取代之環。Heteroaryl: The terms "heteroaryl" and "heteroar-" used alone or as part of a larger moiety (e.g., "heteroaralkyl" or "heteroaralkyloxy") refer to monocyclic or bicyclic groups having 5 to 10 ring atoms (e.g., 5-6 membered monocyclic heteroaryl or 9-10 membered bicyclic heteroaryl); having 6, 10, or 14 pi electrons shared in the cyclic array; and having one to five heteroatoms other than carbon atoms. Heteroaryl groups include, but are not limited to, thienyl, furanyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, isothiazolyl, thiadiazolyl, pyridinyl, oxazinyl, pyrimidinyl, pyrazinyl, indolizinyl, purinyl, naphthyridinyl, pteridinyl, imidazo[1,2-a]pyrimidinyl, imidazo[1,2-a]pyridinyl, imidazo[4,5-b]pyridinyl, imidazo[4,5-c]pyridinyl, pyrrolopyridinyl, pyrrolopyrazinyl, thienopyrimidinyl, triazolopyridinyl, and benzoisoxazolyl. As used herein, the terms "heteroaryl" and "heteroar-" also include groups in which a heteroaromatic ring is fused to one or more aryl, cycloaliphatic, or heterocyclo rings, where the radical or point of attachment is on the heteroaromatic ring (i.e., a bicyclic heteroaryl ring having 1 to 3 heteroatoms). Non-limiting examples include indolyl, isoindolyl, benzothiophenyl, benzofuranyl, dibenzofuranyl, indazolyl, benzimidazolyl, benzotriazolyl, benzothiazolyl, benzothiadiazolyl, benzoxazolyl, quinolinyl, isoquinolinyl, oxazinyl, oxazinyl, quinazolinyl, quinoxalinyl, 4H-quinolizinyl, carbazolyl, acridinyl, phenanthazinyl, phenathiazinyl, phenoxazinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, pyrido[2,3-b]-1,4-oxazin-3(4H )-one, 4H-thieno[3,2-b]pyrrole, and benzoisoxazolyl. The term "heteroaryl" may be used interchangeably with the terms "heteroaryl ring,""heteroaryl," or "heteroaromatic," any of which terms include optionally substituted rings.

雜原子：如本文所用之術語「雜原子」係指氮、氧或硫，且包括氮或硫之任何氧化形式，及鹼性氮之任何四級銨化形式。Impurity atom: As used herein, the term "impurity atom" refers to nitrogen, oxygen, or sulfur, and includes any oxidized form of nitrogen or sulfur, and any quaternary ammonium form of basic nitrogen.

雜環：如本文所用之術語「雜環」、「雜環基」、「雜環基團」及「雜環狀環」可互換使用，且係指穩定的3至8員單環、6至10員二環或10至16員多環雜環部分，其為飽和或部分不飽和的且除碳原子外具有一或多個、諸如一至四個如上文所定義之雜原子。當用於指雜環之環原子時，術語「氮」包括經取代之氮。作為實例，在具有0-3個選自氧、硫或氮之雜原子之飽和或部分不飽和環中，氮可為N (如在3,4-二氫-2H-吡咯基中)、NH (如在吡咯啶基中)或NR⁺(如在N-取代之吡咯啶基中)。雜環狀環可在產生穩定結構之任一雜原子或碳原子處連接至其側基且任一環原子可視情況經取代。該等飽和或部分不飽和雜環基團之實例包括(但不限於)氮雜環丁基、氧雜環丁基、四氫呋喃基、四氫噻吩基、吡咯啶基、六氫吡啶基、十氫喹啉基、噁唑啶基、六氫吡嗪基、二噁烷基、二氧雜環戊烷基、二氮呯基、氧雜氮呯基、硫氮呯基、嗎啉基及硫嗎啉基。雜環基可為單環、二環、三環或多環，較佳地單環、二環或三環，更佳地單環或二環。雙環雜環亦包括如下基團，其中雜環融合至一或多個芳基環。示範性二環雜環基團包括吲哚啉基、異吲哚啉基、苯并二氧雜環戊烯基、1,3-二氫異苯并呋喃基、2,3-二氫苯并呋喃基及四氫喹啉基。二環雜環狀環亦可為螺環系統(例如，除碳原子外具有一或多個如上文所定義之雜原子(例如，一個、兩個、三個或四個雜原子)之7至11員螺環稠合雜環狀環)。二環雜環狀環亦可為橋接環系統(例如，具有一個、兩個或三個橋接原子之7至11員橋接雜環狀環)。Heterocycle: As used herein, the terms "heterocycle,""heterocyclo,""heterocyclogroup," and "heterocyclic ring" are used interchangeably and refer to a stable 3- to 8-membered monocyclic, 6- to 10-membered bicyclic, or 10- to 16-membered polycyclic heterocyclic moiety that is saturated or partially unsaturated and has one or more, such as one to four, heteroatoms as defined above in addition to carbon atoms. The term "nitrogen," when used to refer to a ring atom of a heterocycle, includes substituted nitrogens. For example, in a saturated or partially unsaturated ring having 0-3 heteroatoms selected from oxygen, sulfur, or nitrogen, nitrogen can be N (as in 3,4-dihydro-2H-pyrrolyl), NH (as in pyrrolidinyl), or NR⁺ (as in N-substituted pyrrolidinyl). The heterocyclic ring can be attached to its side group at any heteroatom or carbon atom that creates a stable structure, and any ring atom can be optionally substituted. Examples of such saturated or partially unsaturated heterocyclic groups include, but are not limited to, azacyclobutyl, oxacyclobutyl, tetrahydrofuranyl, tetrahydrothienyl, pyrrolidinyl, hexahydropyridinyl, decahydroquinolinyl, oxazolidinyl, hexahydropyrazinyl, dioxanyl, dioxacyclopentanyl, diazolinyl, oxazolinyl, thiazolinyl, oxazolinyl, oxazolinyl, and thiazolinyl. The heterocyclic group may be monocyclic, bicyclic, tricyclic, or polycyclic, preferably monocyclic, bicyclic, or tricyclic, and more preferably monocyclic or bicyclic. Bicyclic heterocycles also include groups in which the heterocycle is fused to one or more aryl rings. Exemplary bicyclic heterocyclic groups include indolinyl, isoindolinyl, benzodioxolanyl, 1,3-dihydroisobenzofuranyl, 2,3-dihydrobenzofuranyl, and tetrahydroquinolinyl. The bicyclic heterocyclic ring may also be a spirocyclic system (e.g., a 7- to 11-membered spirofused heterocyclic ring having one or more heteroatoms (e.g., one, two, three, or four heteroatoms) as defined above in addition to carbon atoms). The bicyclic heterocyclic ring can also be a bridged ring system (e.g., a 7- to 11-membered bridged heterocyclic ring having one, two, or three bridging atoms).

奈米粒子：如本文所用之術語「奈米粒子」係指小大小、例如最長尺寸通常短於約1000奈米(nm)且通常短於500 nm、或甚至100 nm或更短之離散實體。在許多實施例中，奈米粒子之特徵可在於介於約1 nm與約100 nm之間、或介於約1 μm與約500 nm之間、或介於約1 nm與1000 nm之間的最長尺寸。在許多實施例中，微粒群體之特徵在於低於約1000 nm、約500 nm、約100 nm、約50 nm、約40 nm、約30 nm、約20 nm或約10 nm且通常高於約1 nm之平均大小(例如，最長尺寸)。在許多實施例中，微粒可為實質上球形的(例如，以使得其最長尺寸可為其直徑)。在一些實施例中，奈米粒子之直徑小於100 nm，如由國家衛生研究院(National Institutes of Health)所定義。在一些實施例中，奈米粒子係膠束之原因在於，其包括封閉的隔室，藉由膠束膜與本體溶液分開，通常包括包圍且封閉空間或隔室(例如，以界定內腔)之兩親性實體。在一些實施例中，膠束膜包含至少一種聚合物，例如生物相容性及/或生物可降解聚合物。Nanoparticles: As used herein, the term "nanoparticle" refers to discrete entities of small size, e.g., having a longest dimension typically less than about 1000 nanometers (nm) and often less than 500 nm, or even 100 nm or less. In many embodiments, a nanoparticle can be characterized by a longest dimension between about 1 nm and about 100 nm, or between about 1 μm and about 500 nm, or between about 1 nm and 1000 nm. In many embodiments, a population of microparticles is characterized by an average size (e.g., longest dimension) less than about 1000 nm, about 500 nm, about 100 nm, about 50 nm, about 40 nm, about 30 nm, about 20 nm, or about 10 nm, and typically greater than about 1 nm. In many embodiments, the microparticles can be substantially spherical (e.g., such that their longest dimension can be their diameter). In some embodiments, the nanoparticles have a diameter of less than 100 nm, as defined by the National Institutes of Health. In some embodiments, the nanoparticles are capsules because they include closed compartments separated from the bulk solution by a capsule membrane, typically including an amphiphilic entity that encloses and encloses the space or compartment (e.g. , to define a lumen). In some embodiments, the capsule membrane comprises at least one polymer, such as a biocompatible and/or biodegradable polymer.

核酸/多核苷酸：如本文所用之術語「核酸」係指至少10個或更多個核苷酸之聚合物。在一些實施例中，核酸為或包含DNA。在一些實施例中，核酸係或包含RNA。在一些實施例中，核酸係或包含DNA及RNA之混合物。在一些實施例中，核酸係或包含肽核酸(PNA)。在一些實施例中，核酸為或包含單股核酸。在一些實施例中，核酸係或包含雙股核酸。在一些實施例中，核酸包含單股及雙股部分。在一些實施例中，核酸包含主鏈，該主鏈包含一或多個磷酸二酯鏈接。在一些實施例中，核酸包含主鏈，該主鏈包含磷酸二酯鏈接及非磷酸二酯鏈接。舉例而言，在一些實施例中，核酸可包含主鏈，該主鏈包含一或多個硫代磷酸酯、二硫代磷酸酯、磷醯胺、亞磷酸-硼烷複合物或5'-N-亞磷醯胺鏈接及/或一或多個肽鍵(例如，如在「肽核酸」中)。在一些實施例中，核酸包含一或多個或所有天然殘基(例如，腺嘌呤、胞嘧啶、去氧腺苷、去氧胞苷、去氧鳥苷、去氧胸苷、鳥嘌呤、胸腺嘧啶、尿嘧啶)。在一些實施例中，核酸包含一或多個或所有非天然殘基。在一些實施例中，非天然殘基包含核苷類似物(例如，2-胺基腺苷、2-硫代胸苷、肌苷、吡咯并-嘧啶、3-甲基腺苷、5-甲基胞苷、C-5丙炔基-胞苷、C-5丙炔基-尿苷、2-胺基腺苷、C5-溴尿苷、C5-氟尿苷、C5-碘尿苷、C5-丙炔基-尿苷、C5-丙炔基-胞苷、C5-甲基胞苷、2-胺基腺苷、7-去氮雜腺苷、7-去氮雜鳥苷、8-側氧基腺苷、8-側氧基鳥苷、6-O-甲基鳥嘌呤、2-硫代胞苷、甲基化鹼基、嵌入鹼基及其組合)。在一些實施例中，與天然殘基中之糖相比，非天然殘基包含一或多個經修飾之糖(例如，2'-氟核糖、核糖、2'-去氧核糖、阿拉伯糖及己糖)。在一些實施例中，核酸具有編碼功能基因產物(例如RNA或多肽)之核苷酸序列。在一些實施例中，核酸具有包含一或多個內含子之核苷酸序列。在一些實施例中，核酸可藉由自天然來源分離、酶合成(例如藉由基於互補模板聚合，例如，活體內或活體外)、在重組細胞或系統中繁殖或化學合成來製備。在一些實施例中，核酸之長度係至少3個、4個、5個、6個、7個、8個、9個、10個、15個、20個、25個、30個、35個、40個、45個、50個、55個、60個、65個、70個、75個、80個、85個、90個、95個、100個、110個、120個、130個、140個、150個、160個、170個、180個、190個、20個、225個、250個、275個、300個、325個、350個、375個、400個、425個、450個、475個、500個、600個、700個、800個、900個、1000個、1500個、2000個、2500個、3000個、3500個、4000個、4500個、5000個、5500個、6000個、6500個、7000個、7500個、8000個、8500個、9000個、9500個、10,000個、10,500個、11,000個、11,500個、12,000個、12,500個、13,000個、13,500個、14,000個、14,500個、15,000個、15,500個、16,000個、16,500個、17,000個、17,500個、18,000個、18,500個、19,000個、19,500個或20,000個或更多個殘基或核苷酸。Nucleic Acid/Polynucleotide: As used herein, the term "nucleic acid" refers to a polymer of at least 10 or more nucleotides. In some embodiments, a nucleic acid is or comprises DNA. In some embodiments, a nucleic acid is or comprises RNA. In some embodiments, a nucleic acid is or comprises a mixture of DNA and RNA. In some embodiments, a nucleic acid is or comprises a peptide nucleic acid (PNA). In some embodiments, a nucleic acid is or comprises a single-stranded nucleic acid. In some embodiments, a nucleic acid is or comprises a double-stranded nucleic acid. In some embodiments, a nucleic acid comprises single-stranded and double-stranded portions. In some embodiments, a nucleic acid comprises a backbone comprising one or more phosphodiester linkages. In some embodiments, a nucleic acid comprises a backbone comprising phosphodiester linkages and non-phosphodiester linkages. For example, in some embodiments, a nucleic acid can comprise a backbone comprising one or more phosphorothioate, phosphorodithioate, phosphamide, phosphite-borane complex, or 5'-N-phosphamide linkages and/or one or more peptide bonds (e.g. , as in "peptide nucleic acids"). In some embodiments, a nucleic acid comprises one or more or all naturally occurring residues (e.g. , adenine, cytosine, deoxyadenosine, deoxycytidine, deoxyguanosine, deoxythymidine, guanine, thymine, uracil). In some embodiments, a nucleic acid comprises one or more or all unnatural residues. In some embodiments, the non-natural residue comprises a nucleoside analog (e.g. , 2-aminoadenosine, 2-thiothymidine, inosine, pyrrolo-pyrimidine, 3-methyladenosine, 5-methylcytidine, C-5 propynyl-cytidine, C-5 propynyl-uridine, 2-aminoadenosine, C5-bromouridine, C5-fluorouridine, C5-iodouridine, C5-propynyl-uridine, C5-propynyl-cytidine, C5-methylcytidine, 2-aminoadenosine, 7-deazaadenosine, 7-deazaguanosine, 8-hydroxyadenosine, 8-hydroxyguanosine, 6-O-methylguanine, 2-thiocytidine, methylated bases, embedded bases, and combinations thereof). In some embodiments, the non-natural residue comprises one or more modified sugars (e.g. , 2'-fluororibose, ribose, 2'-deoxyribose, arabinose, and hexose) compared to the sugars in the natural residue. In some embodiments, the nucleic acid has a nucleotide sequence that encodes a functional gene product (e.g., RNA or polypeptide). In some embodiments, the nucleic acid has a nucleotide sequence that comprises one or more introns. In some embodiments, the nucleic acid can be prepared by isolation from a natural source, enzymatic synthesis (e.g., by complementary template-based polymerization,e.g. ,in vivo orin vitro ), propagation in a recombinant cell or system, or chemical synthesis. In some embodiments, the length of the nucleic acid is at least 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 110, 120, 130, 140, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250, 260, 270, 280, 290, 300, 310, 320, 330, 340, 350, 360, 370, 380, 390, 400, 410, 420, 430, 440, 450, 460, 470, 480, 490, 500, 510, 510, 520, 530, 540, 550, 560, 570, 580, 590, 600, 610, 620, 630, 640, 650, 660, 670, 680, 690, 700, 710 , 150, 160, 170, 180, 190, 20, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 500, 600, 700, 800, 900, 1000, 1500, 2000, 250 0, 3000, 3500, 4000, 4500, 5000, 5500, 6000, 6500, 7000, 7500, 8000, 8500, 9000, 9500, 10,000, 10,500, 11,000, 11,500, 12,000, 12,500 0, 13,000, 13,500, 14,000, 14,500, 15,000, 15,500, 16,000, 16,500, 17,000, 17,500, 18,000, 18,500, 19,000, 19,500 or 20,000 or more residues or nucleotides.

核酸粒子：「核酸粒子」可用於將核酸遞送至所關注靶位點(例如，細胞、組織、器官及諸如此類)。核酸粒子可自至少一種陽離子或陽離子可離子化脂質或脂質樣材料及核酸形成。核酸粒子包括基於脂質奈米粒子(LNP)之調配物及基於脂質體複合物(LPX)之調配物。Nucleic Acid Particles: Nucleic acid particles can be used to deliver nucleic acids to target sites of interest (e.g., cells, tissues, organs, and the like). Nucleic acid particles can be formed from at least one cationically or cationically ionizable lipid or lipid-like material and a nucleic acid. Nucleic acid particles include lipid nanoparticle (LNP)-based formulations and liposome complex (LPX)-based formulations.

核苷酸：如本文所用之術語「核苷酸」係指其公認含義。當使用核苷酸數量作為例如多核苷酸大小之指示時，某一數量之核苷酸係指例如多核苷酸之單股上之核苷酸數量。Nucleotide: As used herein, the term "nucleotide" has its generally accepted meaning. When the number of nucleotides is used as an indicator of,for example , the size of a polynucleotide, a certain number of nucleotides refersto the number of nucleotides on a single strand of the polynucleotide.

非經腸：如本文所用之片語「非經腸投與」及「以非經腸方式投與」具有其業內理解之含義，其係指除腸及局部投與外之投與模式，通常藉由注射，且包括(但不限於)靜脈內、肌內、動脈內、鞘內、囊內、眶內、心內、真皮內、腹膜內、經氣管、皮下、表皮下、關節內、囊下、蛛膜下、脊柱內及胸骨內注射及輸注。Parenteral: As used herein, the phrases "parenteral administration" and "administered parenterally" have their meanings as understood in the art and refer to modes of administration other than enteral and topical administration, usually by injection, and include, but are not limited to, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcutaneous, intraarticular, subcapsular, subarachnoid, intraspinal, and intrasternal injection and infusion.

部分不飽和：如本文所用之術語「部分不飽和」係指在環原子之間包括至少一個雙鍵或三鍵之環部分。術語「部分不飽和」意欲涵蓋具有多個不飽和位點之環，但不欲包括芳族(例如，芳基或雜芳基)部分，如本文所定義。Partially Unsaturated: As used herein, the term "partially unsaturated" refers to a ring moiety that includes at least one double or triple bond between ring atoms. The term "partially unsaturated" is intended to encompass rings with multiple sites of unsaturation, but is not intended to include aromatic (e.g., aryl or heteroaryl) moieties, as defined herein.

患者或個體：如本文所用之術語「患者」或「個體」係指投與或可投與所提供組合物，例如用於實驗、診斷、預防、美容及/或治療目的之任何生物體。典型患者或個體包括動物(例如哺乳動物，例如小鼠、大鼠、兔、非人類靈長類動物及/或人類)。在一些實施例中，患者係人類。在一些實施例中，患者或個體患有或易患一或多種病症或疾患。在一些實施例中，患者或個體展示病症或疾患之一或多種症狀。在一些實施例中，患者或個體已經診斷患有一或多種病症或疾患。在一些實施例中，患者或個體正在接受或已接受某些療法以診斷及/或治療疾病、病症或疾患。Patient orSubject: As used herein, the term "patient" or "subject" refers to any organism to which a provided composition is or can be administered, e.g., for experimental, diagnostic, prophylactic, cosmetic, and/or therapeutic purposes. Typical patients or subjects include animals (e.g., mammals, such as mice, rats, rabbits, non-human primates, and/or humans). In some embodiments, the patient is human. In some embodiments, the patient or subject suffers from or is susceptible to one or more conditions or disorders. In some embodiments, the patient or subject exhibits one or more symptoms of a condition or disorder. In some embodiments, the patient or subject has been diagnosed with one or more conditions or disorders. In some embodiments, the patient or subject is receiving or has received certain therapies to diagnose and/or treat a disease, condition, or disorder.

醫藥組合物：如本文所用之術語「醫藥組合物」係指與一或多種醫藥學上可接受之載劑調配在一起之活性劑。在一些實施例中，活性劑係以適於在治療或給藥方案中投與之單位劑量量存在，該治療或給藥方案顯示在投與相關群體時達成預定治療效應之統計學上顯著之概率。在一些實施例中，醫藥組合物可經特殊調配用於以固體或液體形式投與，該固體或液體形式包括適用於以下之彼等形式：口服投與，例如灌服劑(水性或非水性溶液或懸浮液)、錠劑(例如，靶向頰側、舌下及全身吸收之錠劑)、濃注、粉末、顆粒、施用至舌之糊劑；非經腸投與，例如藉由作為例如無菌溶液或懸浮液或持續釋放調配物皮下、肌內、靜脈內或硬膜外注射；局部施用，例如作為施用至皮膚、肺或口腔之乳霜、軟膏或受控釋放貼片或噴霧；陰道內或直腸內，例如作為子宮托、乳霜或泡沫；舌下；眼部；穿真皮；或經鼻、肺部及施用至其他黏膜表面。Pharmaceutical composition: As used herein, the term "pharmaceutical composition" refers to an active agent formulated with one or more pharmaceutically acceptable carriers. In some embodiments, the active agent is present in a unit dosage amount suitable for administration in a treatment or dosing regimen that shows a statistically significant probability of achieving a predetermined therapeutic effect when administered to a relevant population. In some embodiments, the pharmaceutical compositions may be specially formulated for administration in solid or liquid form, including those suitable for oral administration, such as drenches (aqueous or non-aqueous solutions or suspensions), tablets (e.g., tablets targeted to the buccal, sublingual, and systemic absorption), concentrates, powders, granules, pastes for application to the tongue; parenteral administration, For example, by subcutaneous, intramuscular, intravenous, or epidural injection, e.g., as a sterile solution or suspension or sustained-release formulation; topically, e.g., as a cream, ointment, or controlled-release patch or spray for application to the skin, lungs, or mouth; intravaginally or intrarectally, e.g., as a pessary, cream, or foam; sublingually; ophthalmically; transdermally; or nasally, pulmonary, and to other mucosal surfaces.

醫藥學上可接受：如本文所用之片語「醫藥學上可接受」係指在合理醫學判斷範圍內，適用於與人類及動物之組織接觸而無過度毒性、刺激、過敏反應或其他問題或併發症且與合理益處/風險比相稱之彼等化合物、材料、組合物及/或劑量形式。Pharmaceutically acceptable: As used herein, the phrase "pharmaceutically acceptable" refers to those compounds, materials, compositions and/or dosage forms that are, within the scope of sound medical judgment, suitable for use in contact with human and animal tissues without excessive toxicity, irritation, allergic reaction or other problems or complications and are commensurate with a reasonable benefit/risk ratio.

醫藥學上可接受之鹽：如本文所用之術語「醫藥學上可接受之鹽」係指適用於醫藥背景下之該等化合物之鹽，亦即在合理醫學判斷範圍內適用於與人類及低等動物之組織接觸而無過度毒性、刺激、過敏反應及諸如此類且與合理益處/風險比相稱之鹽。醫藥學上可接受之鹽為此項技術中所熟知。舉例而言，S. M. Berge等人在J. Pharmaceutical Sciences, 66: 1-19 (1977)中詳細描述醫藥學上可接受之鹽。Pharmaceutically acceptable salts: As used herein, the term "pharmaceutically acceptable salts" refers to salts of the compounds that are suitable for use in a pharmaceutical setting, i.e., salts that are suitable for use in contact with the tissues of humans and lower animals without excessive toxicity, irritation, allergic reactions, or the like, and that are commensurate with a reasonable benefit/risk ratio, within the scope of sound medical judgment. Pharmaceutically acceptable salts are well known in the art. For example, SM Berge et al. describe pharmaceutically acceptable salts in detail inJ. Pharmaceutical Sciences , 66: 1-19 (1977).

生理條件：如本文所用，具有其業內理解之含義，係指細胞或生物體生活及/或繁殖之條件。在一些實施例中，該術語係指生物體或細胞系統可能存在於自然界中之外部或內部環境之條件。在一些實施例中，生理條件係存在於人類或非人類動物身體內之彼等條件，尤其存在於手術位點處及/或內之彼等條件。生理條件通常包括例如20℃-40℃之溫度範圍、1大氣壓、6-8之pH、1-20 mM之葡萄糖濃度、大氣水準下之氧濃度及其在地球上遇到之重力。在一些實施例中，將實驗室中之條件操縱及/或維持在生理條件下。在一些實施例中，生理條件在生物體中遇到。Physiological conditions: As used herein, the term has its meaning as understood in the art and refers to conditions under which cells or organisms live and/or reproduce. In some embodiments, the term refers to conditions in the external or internal environment of an organism or cellular system as it may exist in nature. In some embodiments, physiological conditions are those conditions that exist within the human or non-human animal body, particularly those conditions that exist at and/or within a surgical site. Physiological conditions typically include, for example, a temperature range of 20°C-40°C, 1 atmosphere of pressure, a pH of 6-8, a glucose concentration of 1-20 mM, oxygen concentrations at atmospheric levels, and gravity as encountered on Earth. In some embodiments, conditions in the laboratory are manipulated and/or maintained at physiological conditions. In some embodiments, physiological conditions are encountered within an organism.

多環：如本文所用之術語「多環」係指具有兩個或更多個環(例如，雜環基環、雜芳基環、環烷基環或芳基環)、具有7至20個原子之飽和或不飽和環系統，其中一或多個碳原子為兩個相鄰環所共有。舉例而言，在一些實施例中，多環系統係指具有三個或更多個環(例如，雜環基環、雜芳基環、環烷基環或芳基環)、具有14至20個原子之飽和或不飽和環系統，其中一或多個碳原子為兩個相鄰環所共有。多環系統中之環可為稠合的(亦即，二環或三環)、螺環或其組合。實例多環係類固醇。Polycyclic: As used herein, the term "polycyclic" refers to a saturated or unsaturated ring system having two or more rings (e.g., a heterocycloalkyl ring, a heteroaryl ring, a cycloalkyl ring, or an aryl ring) with 7 to 20 atoms, wherein one or more carbon atoms are shared by two adjacent rings. For example, in some embodiments, a polycyclic system refers to a saturated or unsaturated ring system having three or more rings (e.g., a heterocycloalkyl ring, a heteroaryl ring, a cycloalkyl ring, or an aryl ring) with 14 to 20 atoms, wherein one or more carbon atoms are shared by two adjacent rings. The rings in a polycyclic system can be fused (ie, bicyclic or tricyclic), spirocyclic, or a combination thereof. Examples of polycyclic rings are steroids.

多肽：如本文所用之術語「多肽」或「肽」通常具有至少三個或更多個胺基酸之聚合物之其公認含義。熟習此項技術者應意識到，術語「多肽」意欲足夠寬泛以不僅涵蓋具有本文所列舉之完整序列之多肽，且亦涵蓋代表該等完整多肽之功能性、生物活性或特徵性片段、部分或結構域(例如，保留至少一種活性之片段、部分或結構域)之多肽。在一些實施例中，多肽可含有L-胺基酸、D-胺基酸或二者皆有，及/或可含有此項技術中已知之多種胺基酸修飾或類似物中之任一者。有用的修飾包括例如末端乙醯化、醯胺化、甲基化等。在一些實施例中，多肽可包含天然胺基酸、非天然胺基酸、合成胺基酸及其組合(例如，可為或包含擬肽物)。Polypeptide: As used herein, the terms "polypeptide" or "peptide" generally have their recognized meaning of a polymer of at least three or more amino acids. Those skilled in the art will appreciate that the term "polypeptide" is intended to be broad enough to encompass not only polypeptides having the complete sequences listed herein, but also polypeptides that represent functional, biologically active, or characteristic fragments, portions, or domains of such complete polypeptides (e.g. , fragments, portions, or domains that retain at least one activity). In some embodiments, a polypeptide may contain L-amino acids, D-amino acids, or both, and/or may contain any of a variety of amino acid modifications or analogs known in the art. Useful modifications include,for example, terminal acetylation, amidation, methylation,and the like . In some embodiments, a polypeptide can comprise natural amino acids, unnatural amino acids, synthetic amino acids, and combinations thereof (eg, can be or comprise a peptidomimetic).

預防(prevent)或預防(prevention)：如本文所用，術語「預防(prevent)」或「預防(prevention)」在與疾病、病症及/或疾患之發生結合使用時，係指降低發展疾病、病症及/或疾患之風險，及/或延遲疾病、病症或疾患之一或多種特徵或症狀之發作。當疾病、病症或疾患之發作已延遲預定時間段時，預防可被視為完成。Prevent orPrevention: As used herein, the term "prevent" or "prevention," when used in conjunction with the occurrence of a disease, disorder, and/or condition, refers to reducing the risk of developing the disease, disorder, and/or condition and/or delaying the onset of one or more characteristics or symptoms of the disease, disorder, or condition. Prevention may be considered accomplished when the onset of the disease, disorder, or condition has been delayed for a predetermined period of time.

參考：如本文所用，描述相對於其進行比較之標準或對照。舉例而言，在一些實施例中，所關注之劑、動物、個體、群體、樣品、序列或值係與參考或對照劑、動物、個體、群體、樣品、序列或值進行比較。在一些實施例中，參考或對照實質上係與所關注之測試或測定同時測試及/或測定。在一些實施例中，參考或對照係歷史參考或對照，視情況地體現在有形媒體中。通常，如熟習此項技術者應理解，參考或對照係在與評價下之條件或情況相當之條件或情況下測定或表徵。熟習此項技術者應瞭解，何時存在足夠相似性以證明依賴於特定可能的參考或對照及/或與特定可能的參考或對照比較係合理的。Reference: As used herein, describes a standard or control relative to which a comparison is made. For example, in some embodiments, an agent, animal, individual, population, sample, sequence, or value of interest is compared to a reference or control agent, animal, individual, population, sample, sequence, or value. In some embodiments, a reference or control is tested and/or assayed substantially simultaneously with the test or assay of interest. In some embodiments, a reference or control is a historical reference or control, optionally embodied in a tangible medium. Generally, as will be understood by those skilled in the art, a reference or control is assayed or characterized under conditions or circumstances equivalent to those under evaluation. Those skilled in the art will understand when sufficient similarity exists to justify reliance on and/or comparison with a particular possible reference or comparison.

核糖核苷酸：如本文所用之術語「核糖核苷酸」涵蓋未經修飾之核糖核苷酸及經修飾之核糖核苷酸。舉例而言，未經修飾之核糖核苷酸包括嘌呤鹼基腺嘌呤(A)及鳥嘌呤(G)，以及嘧啶鹼基胞嘧啶(C)及尿嘧啶(U)。經修飾之核糖核苷酸可包括一或多種修飾，包括(但不限於)例如(a)末端修飾，例如5'末端修飾(例如，磷酸化、去磷酸化、偶聯、反向鏈接等)、3'末端修飾(例如，偶聯、反向鏈接等)，(b)鹼基修飾，例如用經修飾鹼基、穩定鹼基、去穩定鹼基或與擴展的配偶體譜系鹼基配對之鹼基或共軛鹼基替代，(c)糖修飾(例如，在2'位或4'位)或替代糖，及(d)核苷間鏈接修飾，包括修飾或替代磷酸二酯鏈接。術語「核糖核苷酸」亦涵蓋核糖核苷酸三磷酸，包括經修飾及未經修飾之核糖核苷酸三磷酸。Ribonucleotide: As used herein, the term "ribonucleotide" encompasses both unmodified and modified ribonucleotides. For example, unmodified ribonucleotides include the purine bases adenine (A) and guanine (G), and the pyrimidine bases cytosine (C) and uracil (U). Modified ribonucleotides may include one or more modifications, including but not limited to, for example (a) terminal modifications,such as 5' terminal modifications (e.g. , phosphorylation, dephosphorylation, conjugation, reverse linkage, etc. ), 3' terminal modifications (e.g. , conjugation, reverse linkage, etc. ), (b) base modifications,such as replacement with modified bases, stable bases, destabilized bases, or bases that pair with expanded partner bases or conjugated bases, (c) sugar modifications (e.g. , at the 2' or 4' position) or replacement sugars, and (d) internucleoside linkage modifications, including modification or replacement of phosphodiester linkages. The term "ribonucleotide" also encompasses ribonucleotide triphosphates, including modified and unmodified ribonucleotide triphosphates.

核糖核酸(RNA)：如本文所用之術語「RNA」係指核糖核苷酸之聚合物。在一些實施例中，RNA係單股。在一些實施例中，RNA係雙股。在一些實施例中，RNA包含單股及雙股部分。在一些實施例中，RNA可包含如上文「核酸/多核苷酸」之定義中所述之主鏈結構。RNA可為調節RNA (例如，siRNA、microRNA等)或信使RNA (mRNA)。在一些實施例中，其中RNA係mRNA。在其中RNA係mRNA之一些實施例中，RNA在其3’末端通常包含poly(A)區域。在其中RNA係mRNA之一些實施例中，RNA在其5’末端通常包含公認帽結構，例如用於識別mRNA並將mRNA連接至核糖體以起始轉譯。在一些實施例中，RNA為合成RNA。合成RNA包括活體外合成(例如，藉由酶合成方法及/或藉由化學合成方法)之RNA。Ribonucleic acid (RNA): As used herein, the term "RNA" refers to a polymer of ribonucleotides. In some embodiments, the RNA is single-stranded. In some embodiments, the RNA is double-stranded. In some embodiments, the RNA comprises single-stranded and double-stranded portions. In some embodiments, the RNA may comprise a backbone structure as described above in the definition of "nucleic acid/polynucleotide ". The RNA may be a regulatory RNA (e.g. , siRNA, microRNA, etc. ) or a messenger RNA (mRNA). In some embodiments, the RNA is mRNA. In some embodiments in which the RNA is mRNA, the RNA typically comprises a poly(A) region at its 3' end. In some embodiments in which the RNA is mRNA, the RNA typically comprises a recognized cap structure at its 5' end,e.g., for recognizing the mRNA and attaching the mRNA to the ribosome to initiate translation. In some embodiments, the RNA is synthetic RNA. Synthetic RNA includes RNA synthesizedin vitro (eg , by enzymatic synthesis and/or by chemical synthesis).

經取代或視情況經取代：如本文所述，本發明之化合物可含有「視情況經取代」之部分。一般而言，術語「經取代」無論前面是否有術語「視情況地」皆意指，指定部分之一或多個氫經適宜取代基替代。「經取代」適用於自結構(例如，係指至少；且係指至少、或)中明確或隱含之一或多個氫。除非另有指示，否則「視情況經取代」之基團可在基團之每一可取代位置具有適宜取代基，且當任一給定結構中之一個以上之位置可經一個以上之選自所指定基團之取代基取代時，每個位置之取代基可為相同或不同的。本發明所設想之取代基之組合較佳地係可形成穩定的或化學上可行之化合物之取代基組合。如本文所用之術語「穩定」係指當經歷允許其產生、偵測及在某些實施例中其回收、純化及用於本文所提供之一或多個目的之條件時，實質上不會發生變化之化合物。描述為「經取代」之基團較佳地具有1至4個取代基，更佳地1或2個取代基。描述為「視情況經取代」基團可為未經取代或如上文所述「經取代」的。Substituted orOptionally Substituted: As described herein, the compounds of the present invention may contain "optionally substituted" moieties. Generally speaking, the term "substituted," whether preceded by the term "optionally," means that one or more hydrogen atoms in the designated moiety are replaced with a suitable substituent. "Substituted" applies to structures (e.g., Means at least ;and Means at least 、 or ) contains one or more hydrogens, either explicitly or implicitly. Unless otherwise indicated, an "optionally substituted" group may have a suitable substituent at each substitutable position of the group, and when more than one position in any given structure may be substituted with more than one substituent selected from the specified group, the substituents at each position may be the same or different. The combinations of substituents contemplated by the present invention are preferably combinations of substituents that form stable or chemically feasible compounds. The term "stable" as used herein refers to a compound that does not substantially change when subjected to conditions that allow its production, detection, and in some embodiments, its recovery, purification, and use for one or more of the purposes provided herein. Groups described as "substituted" preferably have from 1 to 4 substituents, more preferably 1 or 2 substituents. A group described as "optionally substituted" may be unsubstituted or "substituted" as described above.

「視情況經取代」基團之可取代碳原子上之適宜單價取代基獨立地為鹵素；–(CH₂)_0–4R˚；–(CH₂)_0–4OR˚；-O(CH₂)_0-4R˚、–O–(CH₂)_0–4C(O)OR˚；–(CH₂)_0–4CH(OR˚)₂；–(CH₂)_0–4SR˚；–(CH₂)_0–4Ph，其可經R°取代；–(CH₂)_0–4O(CH₂)_0–1Ph，其可經R°取代；–CH=CHPh，其可經R°取代；–(CH₂)_0–4O(CH₂)_0–1-吡啶基，其可經R°取代；–NO₂；–CN；–N₃；-(CH₂)_0–4N(R˚)₂；–(CH₂)_0–4N(R˚)C(O)R˚；–N(R˚)C(S)R˚；–(CH₂)_0–4N(R˚)C(O)NR˚₂；-N(R˚)C(S)NR˚₂；–(CH₂)_0–4N(R˚)C(O)OR˚；-N(R˚)N(R˚)C(O)R˚；-N(R˚)N(R˚)C(O)NR˚₂；-N(R˚)N(R˚)C(O)OR˚；–(CH₂)_0–4C(O)R˚；C(S)R˚；–(CH₂)_0–4C(O)OR˚；–(CH₂)_0–4C(O)SR˚；-(CH₂)_0–4C(O)OSiR˚₃；–(CH₂)_0–4OC(O)R˚；–OC(O)(CH₂)_0–4SR°；–(CH₂)_0–4SC(O)R˚；–(CH₂)_0–4C(O)NR˚₂；–C(S)NR˚₂；–C(S)SR°；–SC(S)SR°, -(CH₂)_0–4OC(O)NR˚₂；-C(O)N(OR˚)R˚；–C(O)C(O)R˚；–C(O)CH₂C(O)R˚；–C(NOR˚)R˚；-(CH₂)_0–4SSR˚；–(CH₂)_0–4S(O)₂R˚；–(CH₂)_0–4S(O)₂OR˚；–(CH₂)_0–4OS(O)₂R˚；–S(O)₂NR˚₂；-(CH₂)_0–4S(O)R˚；-N(R˚)S(O)₂NR˚₂；–N(R˚)S(O)₂R˚；–N(OR˚)R˚；–C(NH)NR˚₂；–P(O)₂R˚；-P(O)R˚₂；-OP(O)R˚₂；–OP(O)(OR˚)₂；SiR˚₃；–(C_1–4直鏈或分枝伸烷基)O–N(R˚)₂；或–(C_1–4直鏈或分枝伸烷基)C(O)O–N(R˚)₂，其中各R˚可如下文所定義經取代且獨立地為氫、C_1–6脂族基、–CH₂Ph、–O(CH₂)_0–1Ph、-CH₂-(5至6員雜芳基環)，或具有0-4個獨立地選自氮、氧或硫之雜原子之3至6員飽和、部分不飽和或芳基環，或儘管具有上文定義，兩個獨立出現之R˚與其間插原子一起形成具有0-4個獨立地選自氮、氧或硫之雜原子之3至12員飽和、部分不飽和或芳基單環或雙環，其可如下文所定義經取代。Suitable monovalent substituents on the substitutable carbon atom of the "optionally substituted" group are independently halogen; –(CH₂ )_0–4 R°; –(CH₂ )_0–4 OR°; –O(CH₂ )_0–4 R°, –O–(CH₂ )_0–4 C(O)OR°; –(CH₂ )_0–4 CH(OR°)₂ ; –(CH₂ )_0–4 SR°; –(CH₂ )_0–4 Ph, which may be substituted by R°; –(CH₂ )_0–4 O(CH₂ )_0–1 Ph, which may be substituted by R°; –CH=CHPh, which may be substituted by R°; –(CH₂ )_0–4 O(CH₂ )_0–1 -pyridyl, which may be substituted by R°; –NO₂ ; –CN; –N₃ ;-(CH₂ )_0–4 N(R˚)₂ ;–(CH₂ )_0–4 N(R˚)C(O)R˚;–N(R˚)C(S)R˚;–(CH₂ )_0–4 N(R˚)C(O)NR˚₂ ;-N(R˚)C(S)NR˚₂ ;–(CH₂ )_0–4 N(R˚)C(O)OR˚;-N(R˚)N(R˚)C(O)R˚;-N(R˚)N(R˚)C(O)NR˚₂ ;-N(R˚)N(R˚)C(O)OR˚;–(CH₂ )_0–4 C(O)R˚;C(S)R˚;–(CH₂ )_0–4 C(O)OR˚;–(CH₂ )_0–4 C(O)SR˚;-(CH₂ )_0–4 C(O)OSiR˚₃ ;–(CH₂ )_0–4 OC(O)R˚;–OC(O)(CH₂ )_0–4 SR°;–(CH₂ )_0–4 SC(O)R˚;–(CH₂ )_0–4 C(O)NR˚₂ ;–C(S)NR˚₂ ;–C(S)SR°;–SC(S)SR°, -(CH₂ )_0–4 OC(O)NR˚₂ ;-C(O)N(OR˚)R˚;–C(O)C(O)R˚;–C(O)CH₂ C(O)R˚;–C(NOR˚)R˚;-(CH₂ )_0–4 SSR˚;– (CH₂ )_0–4 S(O)₂ R˚;–(CH₂ )_0–4 S(O)₂ OR˚; –(CH₂ )_0–4 OS(O)₂ R˚; –S(O)₂ NR˚₂ ; –(CH₂ )_0–4 S(O)R˚; –N(R˚)S(O)₂ NR˚₂ ; –N(R˚)S(O)₂ R˚; –N(OR˚)R˚; –C(NH)NR˚₂ ; –P(O)₂ R˚; –P(O)R˚₂ ; –OP(O)R˚₂ ; –OP(O)(OR˚)₂ ; SiR˚₃ ; –(C_1–4 straight chain or branched alkylene)O–N(R˚)₂ ; or –(C_1–4 straight chain or branched alkylene)C(O)O–N(R˚)₂ , wherein each R˚ may be substituted as defined below and is independently hydrogen, C_1-6 membered aliphatic, -CH₂ Ph, -O(CH₂ )_0-1 Ph, -CH₂ -(5- to 6-membered heteroaryl ring), or a 3- to 6-membered saturated, partially unsaturated or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur, or notwithstanding the above definition, two independent occurrences of R˚ together with their intervening atoms form a 3- to 12-membered saturated, partially unsaturated or aryl monocyclic or bicyclic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur, which may be substituted as defined below.

R° (或藉由兩個獨立出現之R°與其間插原子一起形成之環)上之適宜單價取代基獨立地為鹵素、–(CH₂)_0–2R^●、-(鹵基R^●)、–(CH₂)_0–2OH、–(CH₂)_0–2OR^●、–(CH₂)_0–2CH(OR^●)₂、-O(鹵基R^●)、-CN、-N₃、–(CH₂)_0–2C(O)R^●、–(CH₂)_0–2C(O)OH、–(CH₂)_0–2C(O)OR^●、–(CH₂)_0–2SR^●、–(CH₂)_0–2SH、–(CH₂)_0–2NH₂、–(CH₂)_0–2NHR^●、–(CH₂)_0–2NR^●₂、–NO₂、–SiR^●₃、–OSiR^●₃、‑C(O)SR^●、-(C_1–4直鏈或分枝伸烷基)C(O)OR^●或-SSR^●，其中各R^●未經取代或在前面有「鹵基」之情況下僅經一或多個鹵素取代，且獨立地選自C_1–4脂族基、-CH₂Ph、-O(CH₂)_0-1Ph或具有0-4個獨立地選自氮、氧或硫之雜原子之3至6員飽和、部分不飽和或芳基環。R˚之飽和碳原子上之適宜二價取代基包括=O及=S。Suitable monovalent substituents on R° (or the ring formed by two independently occurring R° together with the intervening atoms) are independently halogen, –(CH₂ )_0–2 R^● , –(halogen R^● ), –(CH₂ )_0–2 OH, –(CH₂ )_0–2 OR^● , –(CH₂ )_0–2 CH(OR^● )₂ , –O(halogen R^● ), –CN, –N₃ , –(CH₂ )_0–2 C(O)R^● , –(CH₂ )_0–2 C(O)OH, –(CH₂ )_0–2 C(O)OR^● , –(CH₂ )_0–2 SR^● , –(CH₂ )_0–2 SH, –(CH₂ )_0–2 NH₂ , –(CH₂ )_0-2 NHR^● , –(CH₂ )_0-2 NR^●₂ , –NO₂ , –SiR^●₃ , –OSiR^●₃ , –C(O)SR^● , -(C_1-4 straight-chain or branched alkylene)C(O)OR^● , or -SSR^● , wherein each R^● is unsubstituted or, when preceded by “halogen”, is substituted only with one or more halogens, and is independently selected from C_1-4 aliphatic, -CH₂ Ph, -O(CH₂ )_0-1 Ph, or a 3- to 6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. Suitable divalent substituents on the saturated carbon atom of R˚ include =O and =S.

「視情況經取代」基團之飽和碳原子上之適宜二價取代基包括以下：=O (「側氧基」)、=S、=NNR^*₂、=NNHC(O)R^*、=NNHC(O)OR^*、=NNHS(O)₂R^*、=NR^*、=NOR^*、-O(C(R^*₂))_2-3O-或-S(C(R^*₂))_2-3S-，其中各獨立出現之R^*選自氫、可如下文所定義經取代之C_1-6脂族，或具有0-4個獨立地選自氮、氧或硫之雜原子之未經取代的5至6員飽和、部分不飽和或芳基環。結合至「視情況經取代」之基團之鄰位可取代碳之適宜二價取代基包括：-O(CR^*₂)_2-3O-，其中每一獨立出現之R^*選自氫、可如下文所定義經取代之C_1-6脂族，或未經取代之具有0-4個獨立地選自氮、氧或硫之雜原子之5-6員飽和、部分不飽和或芳基環。Suitable divalent substituents on a saturated carbon atom of an "optionally substituted" group include the following: =O ("oxo"), =S, =NNR^*₂ , =NNHC(O)R^* , =NNHC(O)OR^* , =NNHS(O)_2R^* , =NR^* , =NOR^* , -O(C(R^*₂ ))_2-3O- , or -S(C(R^*₂ ))_2-3S- , wherein each independent occurrence of R^* is selected from hydrogen, a_C1-6 aliphatic which may be substituted as defined below, or an unsubstituted 5- to 6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. Suitable divalent substituents attached to an vicinal substitutable carbon of the "optionally substituted" group include: -O(CR^*₂ )_2-3 O-, wherein each independent occurrence of R^* is selected from hydrogen, a C_1-6 aliphatic group which may be substituted as defined below, or an unsubstituted 5-6 membered saturated, partially unsaturated or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur.

R^*之脂族基團上之適宜取代基包括鹵素、-R^●、-(鹵基R^●)、-OH、-OR^●、-O(鹵基R^●)、-CN、-C(O)OH、-C(O)OR^●、-NH₂、-NHR^●、-NR^●₂或-NO₂，其中各R^●未經取代或在前面有「鹵基」之情況下僅經一或多個鹵素取代，且獨立地為C_1-4脂族、-CH₂Ph、-O(CH₂)_0-1Ph，或具有0-4個獨立地選自氮、氧或硫之雜原子之3至6員飽和、部分不飽和或芳基環。Suitable substituents on the aliphatic group of R^* include halogen, -R^● , -(halogen R^● ), -OH, -OR^● , -O(halogen R^● ), -CN, -C(O)OH, -C(O)OR^● ,_-NH2 , -NHR^● , -NR^●₂ or_-NO2 , wherein each R^● is unsubstituted or, if preceded by "halogen", is substituted only by one or more halogens, and is independently_C1-4 aliphatic,_-CH2Ph , -O(_CH2 )_0-1Ph , or a 3- to 6-membered saturated, partially unsaturated or aromatic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur.

「視情況經取代」基團之可取代氮上之適宜取代基包括-R^†、-NR^†₂、-C(O)R^†、-C(O)OR^†、-C(O)C(O)R^†、-C(O)CH₂C(O)R^†、-S(O)₂R^†、-S(O)₂NR^†₂、-C(S)NR^†₂、-C(NH)NR^†₂，或-N(R^†)S(O)₂R^†；其中各R^†獨立地為氫、可如下文所定義經取代之C_1-6脂族、未經取代之-OPh，或具有0-4個獨立地選自氮、氧或硫之雜原子之未經取代的3至6員飽和、部分不飽和或芳基環，或儘管具有上文定義，兩個獨立出現之R^†與其間插原子一起形成具有0-4個獨立地選自氮、氧或硫之雜原子之未經取代的3至12員飽和、部分不飽和或芳基單環或雙環。Suitable substituents on the substitutable nitrogen of an "optionally substituted" group include -R^† , -NR^†₂ , -C(O)R^† , -C(O)OR^† , -C(O)C(O)R^† , -C(O)CH₂ C(O)R † , -S(O) 2 R^† , -S(O)₂ NR^{† 2}_, -C(S)NR^†₂ , -C(NH)NR^†₂ , or -N(R^† )_S (^O )₂ R^† ; wherein each R^† is independently hydrogen, C(O) which may be substituted as defined below, or C(O) which may be substituted as defined below._1-6 aliphatic, unsubstituted -OPh, or an unsubstituted 3- to 6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or, notwithstanding the above definition, two independent occurrences of R^† together with their intervening atoms form an unsubstituted 3- to 12-membered saturated, partially unsaturated, or aryl monocyclic or bicyclic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.

R^†之脂族基團上之適宜取代基獨立地為鹵素、-R^●、-(鹵基R^●)、-OH、-OR^●、-O(鹵基R^●)、-CN、-C(O)OH、-C(O)OR^●、-NH₂、-NHR^●、-NR^●₂或-NO₂，其中各R^●未經取代或在前面有「鹵基」之情況下僅經一或多個鹵素取代，且獨立地為C_1-4脂族、-CH₂Ph、-O(CH₂)_0-1Ph，或具有0-4個獨立地選自氮、氧或硫之雜原子之3至6員飽和、部分不飽和或芳基環。Suitable substituents on the aliphatic group of R^† are independently halogen, -R^● , -(halogen R^● ), -OH, -OR^● , -O(halogen R^● ), -CN, -C(O)OH, -C(O)OR^● , -NH₂ , -NHR^● , -NR^●₂ or -NO₂ , wherein each R^● is unsubstituted or, if preceded by "halogen", is substituted only by one or more halogens, and is independently C_1-4 aliphatic, -CH₂ Ph, -O(CH₂ )_0-1 Ph, or a 3- to 6-membered saturated, partially unsaturated or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur.

小分子：如本文所用之術語「小分子」意指低分子量有機及/或無機化合物。一般而言，「小分子」係大小小於約5千道爾頓(kD)之分子。在一些實施例中，小分子小於約4 kD、3 kD、約2 kD或約1 kD。在一些實施例中，小分子小於約800道爾頓(D)、約600 D、約500 D、約400 D、約300 D、約200 D或約100 D。在一些實施例中，小分子小於約2000 g/mol、小於約1500 g/mol、小於約1000 g/mol、小於約800 g/mol或小於約500 g/mol。在一些實施例中，小分子不為聚合物。Small molecule: As used herein, the term "small molecule" refers to low molecular weight organic and/or inorganic compounds. Generally, a "small molecule" is a molecule less than about 5 kilodaltons (kD). In some embodiments, a small molecule is less than about 4 kD, 3 kD, about 2 kD, or about 1 kD. In some embodiments, a small molecule is less than about 800 daltons (D), about 600 D, about 500 D, about 400 D, about 300 D, about 200 D, or about 100 D. In some embodiments, a small molecule is less than about 2000 g/mol, less than about 1500 g/mol, less than about 1000 g/mol, less than about 800 g/mol, or less than about 500 g/mol. In some embodiments, the small molecule is not a polymer.

在一些實施例中，小分子不包括聚合物部分。在一些實施例中，小分子不為及/或不包含蛋白質或多肽(例如不為寡肽或肽)。在一些實施例中，小分子不為及/或不包含多核苷酸(例如不為寡核苷酸)。在一些實施例中，小分子不為及/或不包含多糖；例如，在一些實施例中，小分子不為糖蛋白、蛋白多糖、糖脂等。在一些實施例中，小分子不為脂質。In some embodiments, the small molecule does not include a polymer moiety. In some embodiments, the small molecule is not and/or does not comprise a protein or polypeptide (e.g., is not an oligopeptide or peptide). In some embodiments, the small molecule is not and/or does not comprise a polynucleotide (e.g., is not an oligonucleotide). In some embodiments, the small molecule is not and/or does not comprise a polysaccharide; for example, in some embodiments, the small molecule is not a glycoprotein, proteoglycan, glycolipid,etc. In some embodiments, the small molecule is not a lipid.

在一些實施例中，小分子係調節劑(例如係抑制劑或活化劑)。在一些實施例中，小分子為生物活性劑。在一些實施例中，小分子係可偵測的(例如包含至少一個可偵測部分)。在一些實施例中，小分子係治療劑。In some embodiments, the small molecule is a modulator (e.g., an inhibitor or activator). In some embodiments, the small molecule is a bioactive agent. In some embodiments, the small molecule is detectable (e.g., comprises at least one detectable moiety). In some embodiments, the small molecule is a therapeutic agent.

閱讀本揭示案之熟習此項技術者應意識到，本文所述之某些小分子化合物可以多種形式(例如晶體形式(例如，多形體、溶劑合物等)、鹽形式、受保護形式、前藥形式、酯形式、異構形式(例如，光學及/或結構異構物)、同位素形式等)中之任一者提供及/或使用。Those skilled in the art who read this disclosure will appreciate that certain small molecule compounds described herein may be provided and/or used in any of a variety of forms, such as crystalline forms (e.g., polymorphs, solvates, etc.), salt forms, protected forms, prodrug forms, ester forms, isomeric forms (e.g., optical and/or structural isomers), isotopic forms, etc.

熟習此項技術者應意識到，某些小分子化合物具有可以一或多種立體異構形式存在之結構。在一些實施例中，根據本揭示案，該小分子可以個別鏡像異構物、非鏡像異構物或幾何異構物之形式使用，或可呈立體異構物之混合物形式；在一些實施例中，根據本揭示案，該小分子可以外消旋混合物形式使用。Those skilled in the art will appreciate that certain small molecule compounds have structures that can exist in one or more stereoisomers. In some embodiments, according to the present disclosure, the small molecules can be used as individual mirror isomers, non-mirror isomers, or geometric isomers, or as a mixture of stereoisomers; in some embodiments, according to the present disclosure, the small molecules can be used as racemic mixtures.

熟習此項技術者應意識到，某些小分子化合物具有可以一或多種互變異構形式存在之結構。在一些實施例中，根據本揭示案，該小分子可以個別互變異構物之形式、或以在互變異構形式之間相互轉化之形式使用。Those skilled in the art will appreciate that certain small molecule compounds have structures that can exist in one or more tautomeric forms. In some embodiments, according to the present disclosure, the small molecules can be used in the form of individual tautomers or in a form that is interconvertible between tautomeric forms.

熟習此項技術者應意識到，某些小分子化合物具有容許同位素取代(例如，²H或³H取代H；¹¹C、¹³C或¹⁴C取代¹²C；¹³N或¹⁵N取代¹⁴N；¹⁷O或¹⁸O取代¹⁶O；³⁶Cl取代³⁵Cl或³⁷Cl；⁸F取代¹⁹F；¹³¹I取代¹²⁷I等)之結構。在一些實施例中，根據本揭示案，該小分子可以一或多種同位素修飾形式或其混合物使用。Those skilled in the art will appreciate that certain small molecule compounds have structures that allow for isotopic substitution (e.g.,^2H or^3H for H;^11C ,^13C , or^14C for^12C ;^13N or^15N for^14N ;^17O or^18O for^16O ;^36Cl for^35Cl or^37Cl ;^8F for^19F ;^131I for^127I , etc.). In some embodiments, the small molecules disclosed herein can be used in one or more isotopically modified forms or mixtures thereof.

在一些實施例中，提及特定小分子化合物可能指該化合物之具體形式。在一些實施例中，特定小分子化合物可以鹽形式(例如，以酸加成鹽或鹼加成鹽形式，此端視化合物而定)提供及/或使用；在一些此類實施例中，鹽形式可為醫藥學上可接受之鹽形式。In some embodiments, reference to a particular small molecule compound may refer to a specific form of the compound. In some embodiments, a particular small molecule compound may be provided and/or used in salt form (e.g., as an acid addition salt or a base addition salt, depending on the compound); in some such embodiments, the salt form may be a pharmaceutically acceptable salt form.

在一些實施例中，根據本揭示案，當小分子化合物係在自然界中存在或發現之化合物時，該化合物可以與其在自然界中存在或發現不同之形式提供及/或使用。熟習此項技術者應瞭解，在一些實施例中，特定小分子化合物之製劑(含有絕對或相對量之化合物或其特定形式，其不同於所關注參考製劑中(例如，來自所關注來源、例如生物或環境來源之初級樣品中)存在之絕對或相對(相對於製劑之另一組分，包括例如化合物之另一形式)量之化合物或形式)不同於在參考製劑或來源中存在之化合物。因此，在一些實施例中，例如，小分子化合物之單一立體異構物之製劑可視為不同於化合物之外消旋混合物之化合物形式；小分子化合物之特定鹽可視為不同於化合物之另一鹽形式之形式；僅含含有雙鍵之一種構形異構物((Z)或(E))之化合物形式之製劑可視為不同於含有雙鍵之另一構形異構物((E)或(Z))之化合物形式之化合物形式；其中一或多個原子係不同於參考製劑中存在之同位素的製劑可視為不同形式等。In some embodiments, according to the present disclosure, when a small molecule compound is a compound that exists or is found in nature, the compound can be provided and/or used in a form that is different from that in which it exists or is found in nature. Those skilled in the art will understand that in some embodiments, a formulation of a particular small molecule compound (containing an absolute or relative amount of the compound or a particular form thereof that is different from the absolute or relative (relative to another component of the formulation, including, for example, another form of the compound) amount present in a reference formulation of interest (e.g., a primary sample from a source of interest, such as a biological or environmental source)) is different from the compound present in the reference formulation or source. Thus, in some embodiments, for example, a preparation of a single stereoisomer of a small molecule compound can be considered a different form of the compound from a racemic mixture of the compound; a particular salt of a small molecule compound can be considered a different form from another salt form of the compound; a preparation containing only a form of the compound containing one conformational isomer ((Z) or (E)) of a double bond can be considered a different form of the compound from a form of the compound containing another conformational isomer ((E) or (Z)) of the double bond; a preparation in which one or more atoms are isotopically different from that present in a reference preparation can be considered a different form, etc.

熟習此項技術者應進一步瞭解，在小分子結構中，如本文所用之符號係指兩個原子之間的連接點。另外或替代地，符號係指呈螺環方式之環之連接點。Those skilled in the art will further understand that in small molecule structures, the symbols used herein are Refers to the point of connection between two atoms. Additionally or alternatively, the symbol Refers to the connection point of the ring in the form of a screw ring.

治療：如本文所用之術語「治療(treat)」、「治療(treatment)」或「治療(treating)」係指用於部分或完全緩和、改善、減輕、抑制、預防疾病、病症及/或疾患之一或多個症狀或特徵、延遲其發作、降低其嚴重程度及/或減小其發生率的任一方法。治療可投與未展現疾病、病症及/或疾患之徵象之個體。在一些實施例中，治療可投與僅展現疾病、病症及/或疾患之早期徵象之個體，例如用於降低患上與疾病、病症及/或疾患相關之病狀風險之目的。Treatment: As used herein, the terms "treat,""treatment," or "treating" refer to any method used to partially or completely alleviate, ameliorate, alleviate, suppress, prevent, delay onset, reduce severity, and/or reduce the incidence of one or more symptoms or features of a disease, disorder, and/or condition. Treatment can be administered to a subject who does not exhibit signs of the disease, disorder, and/or condition. In some embodiments, treatment can be administered to a subject who exhibits only early signs of the disease, disorder, and/or condition, for example, to reduce the risk of developing conditions associated with the disease, disorder, and/or condition.

陽離子或可離子化脂質本揭示案尤其提供陽離子或可離子化脂質。本文所述之化合物亦稱為「可離子化」或「陽離子」脂質。此類脂質意欲意指在一些實施例中能夠變得小於生理pH之陽離子(亦即，變得帶正電荷)之化合物。在一些實施例中，本文所述之實例脂質在小於7、小於6.5、小於6、小於5.5、小於5、小於4.5或小於4之pH下變得帶正電荷。在一些實施例中，包含複數種本文所述之陽離子或可離子化脂質之組合物的特徵在於該組合物中10%或更多之陽離子或可離子化脂質包含正電荷。Cationic or Ionizable Lipids The present disclosure provides, inter alia, cationic or ionizable lipids. The compounds described herein are also referred to as "ionizable" or "cationic" lipids. Such lipids are intended to refer to compounds that, in some embodiments, are capable of becoming cationic (i.e., becoming positively charged) at less than physiological pH. In some embodiments, the example lipids described herein become positively charged at a pH of less than 7, less than 6.5, less than 6, less than 5.5, less than 5, less than 4.5, or less than 4. In some embodiments, a composition comprising a plurality of cationic or ionizable lipids described herein is characterized in that 10% or more of the cationic or ionizable lipids in the composition comprise a positive charge.

在一些實施例中，本揭示案提供式I化合物：I 或其醫藥學上可接受之鹽，其中 M¹及M²各自獨立地選自-(CH₂)_p1-； L¹及L²各自選自鍵、-OC(O)-或-C(O)O-； T¹及T²各自為：； L³為視情況經取代之C₁-C₆脂族基； G¹為-OH或-N(R^a)₂，且 R^a為H、視情況經取代之C₁-C₆脂族基或視情況經取代之C₃-C₆環脂族基； p1為選自2-20之整數，包括端值； p2及p3各自為選自1-20之整數，包括端值，且p2及p3係相同的。In some embodiments, the present disclosure provides compounds of Formula I: I or a pharmaceutically acceptable salt thereof, wherein^M1 and^M2 are each independently selected from -(_CH2 )_p1- ;^L1 and^L2 are each selected from a bond, -OC(O)- or -C(O)O-;^T1 and^T2 are each:^L3 is an optionally substituted_C1 -_C6 aliphatic group;^G1 is -OH or -N(^Ra )₂ , and^Ra is H, an optionally substituted_C1 -_C6 aliphatic group, or an optionally substituted_C3 -_C6 cycloaliphatic group; p1 is an integer selected from 2 to 20, inclusive; p2 and p3 are each an integer selected from 1 to 20, inclusive, and p2 and p3 are the same.

在一些實施例中，當部分T¹及T²相同且其中p2及p3中之每一者在式I化合物中相同時，本揭示案認識到令人驚訝之益處。In some embodiments, the present disclosure recognizes surprising benefits when moieties^T1 and^T2 are identical and wherein each of p2 and p3 are identical in a compound of Formula I.

如本文所述，M¹及M²各自獨立地選自-(CH₂)_p1-。在一些實施例中，M¹及M²係相同的。在一些實施例中，T¹及T²係不同的。在一些實施例中，M¹係-(CH₂)_2-10-。在一些實施例中，M¹係-(CH₂)_4-6-。在一些實施例中，M¹係-(CH₂)₄-。在一些實施例中，M¹係-(CH₂)₅-。在一些實施例中，M¹係-(CH₂)₆-。As described herein,^M1 and^M2 are each independently selected from -(_CH2 )_p1- . In some embodiments,^M1 and^M2 are the same. In some embodiments,^T1 and^T2 are different. In some embodiments,^M1 is -(_CH2 )_2-10- . In some embodiments,^M1 is -(_CH2 )_4-6- . In some embodiments,^M1 is -(_CH2 )_4- . In some embodiments,^M1 is -(_CH2 )_5- . In some embodiments,^M1 is -(_CH2 )_6- .

在一些實施例中，M²係-(CH₂)_2-10-。在一些實施例中，M²係-(CH₂)_4-6-。在一些實施例中，M²係-(CH₂)₄-。在一些實施例中，M²係-(CH₂)₅-。在一些實施例中，M²係-(CH₂)₆-。In some embodiments, M² is -(CH₂ )_2-10 -. In some embodiments, M² is -(CH₂ )_4-6 -. In some embodiments, M² is -(CH₂ )₄ -. In some embodiments, M² is -(CH₂ )₅ -. In some embodiments, M² is -(CH₂ )₆ -.

。在一些實施例中，M¹及M²各自係-(CH₂)_2-10-。在一些實施例中，M¹及M²各自係–(CH₂)_4-6-。在一些實施例中，M¹及M²各自係–(CH₂)₄-。在一些實施例中，M¹及M²各自係–(CH₂)₅-。在一些實施例中，M¹及M²各自係–(CH₂)₆-。In some embodiments,^M1 and^M2 are each -(CH₂ )_2-10 -. In some embodiments,^M1 and^M2 are each -(CH₂ )_4-6 -. In some embodiments,^M1 and^M2 are each -(CH₂ )₄ -. In some embodiments,^M1 and^M2 are each -(CH₂ )₅ -. In some embodiments,^M1 and^M2 are each -(CH₂ )₆ -.

如本文所述，p1為選自2-20之整數，包括端值。在一些實施例中，p1為選自2-10之整數，包括端值。在一些實施例中，p1為2、3、4、5、6、7、8、9或10。在一些實施例中，p1為選自4、5或6之整數。As described herein, p1 is an integer selected from 2-20, inclusive. In some embodiments, p1 is an integer selected from 2-10, inclusive. In some embodiments, p1 is 2, 3, 4, 5, 6, 7, 8, 9, or 10. In some embodiments, p1 is an integer selected from 4, 5, or 6.

如本文所述，L¹及L²各自選自鍵、-OC(O)-及-C(O)O-。在一些實施例中，L¹及L²係相同的。在一些實施例中，L¹及L²係不同的。在一些實施例中，L¹及L²選自-OC(O)-及-C(O)O-。在一些實施例中，L¹係鍵。在一些實施例中，L¹係-OC(O)-。在一些實施例中，L¹係-C(O)O-。在一些實施例中，L²係鍵。在一些實施例中，L²係-OC(O)-。在一些實施例中，L²係-C(O)O-。在一些實施例中，L¹及L²各自係鍵。在一些實施例中，L¹及L²各自係-OC(O)-。在一些實施例中，L¹及L²各自係-C(O)O-。As described herein,^L1 and^L2 are each selected from a bond, -OC(O)-, and -C(O)O-. In some embodiments,^L1 and^L2 are the same. In some embodiments,^L1 and^L2 are different. In some embodiments,^L1 and^L2 are selected from -OC(O)- and -C(O)O-. In some embodiments,^L1 is a bond. In some embodiments,^L1 is -OC(O)-. In some embodiments,^L1 is -C(O)O-. In some embodiments,^L2 is a bond. In some embodiments,^L2 is -OC(O)-. In some embodiments,^L2 is -C(O)O-. In some embodiments,^L1 and^L2 are each a bond. In some embodiments,^L1 and^L2 are each -OC(O)-. In some embodiments, L¹ and L² are each -C(O)O-.

如本文所述，T¹及T²各自為：。As described herein,^T1 and^T2 are each: .

在一些實施例中，T¹及T²係相同的。In some embodiments,^T1 and^T2 are the same.

如本文所述，p2及p3各自為選自1-20之整數，包括端值，且p2及p3係相同的。在一些實施例中，p2及p3各自為選自3-9之整數，包括端值。在一些實施例中，p2及p3各自為3、4、5、6、7、8或9。在一些實施例中，p2及p3各自為3。在一些實施例中，p2及p3各自為4。在一些實施例中，p2及p3各自為5。在一些實施例中，p2及p3各自為6。在一些實施例中，p2及p3各自為7。在一些實施例中，p2及p3各自為8。在一些實施例中，p2及p3各自為9。在一些實施例中，p2及p3各自為4、5或6。As described herein, p2 and p3 are each an integer selected from 1-20, inclusive, and p2 and p3 are the same. In some embodiments, p2 and p3 are each an integer selected from 3-9, inclusive. In some embodiments, p2 and p3 are each 3, 4, 5, 6, 7, 8, or 9. In some embodiments, p2 and p3 are each 3. In some embodiments, p2 and p3 are each 4. In some embodiments, p2 and p3 are each 5. In some embodiments, p2 and p3 are each 6. In some embodiments, p2 and p3 are each 7. In some embodiments, p2 and p3 are each 8. In some embodiments, p2 and p3 are each 9. In some embodiments, p2 and p3 are each 4, 5, or 6.

在一些實施例中，T¹及T²各自選自：、、、、及。In some embodiments,^T1 and^T2 are each selected from: 、 、 、 、 and .

在一些實施例中，-M¹-L¹-T¹中之碳原子之總數介於20-30之間，包括端值。在一些實施例中，-M¹-L¹-T¹由以下表示：In some embodiments, the total number of carbon atoms in -M¹ -L¹ -T¹ is between 20 and 30, inclusive. In some embodiments, -M¹ -L¹ -T¹ is represented by:

其中p2及p3各自為選自2至10之整數，包括端值。在一些實施例中，-M¹-L¹-T¹由以下表示：其中p2及p3各自選自4、5或6。Wherein p2 and p3 are each integers selected from 2 to 10, inclusive. In some embodiments, -M¹ -L¹ -T¹ is represented by: Wherein p2 and p3 are each selected from 4, 5 or 6.

在一些實施例中，-M²-L²-T²中之碳原子之總數介於20-30之間，包括端值。在一些實施例中，-M²-L²-T²由以下表示：其中p2及p3各自為選自2至10之整數，包括端值。在一些實施例中，-M²-L²-T²由以下表示：其中p2及p3各自為4、5或6。In some embodiments, the total number of carbon atoms in -M² -L² -T² is between 20 and 30, inclusive. In some embodiments, -M² -L² -T² is represented by: Wherein p2 and p3 are each integers selected from 2 to 10, inclusive. In some embodiments, -M² -L² -T² is represented by: wherein p2 and p3 are 4, 5 or 6 respectively.

在一些實施例中，M¹-L¹-T¹及M²-L²-T²係相同的。In some embodiments, M¹ -L¹ -T¹ and M² -L² -T² are the same.

在一些實施例中，M¹-L¹-T¹及M²-L²-T²各自選自、、、、、、、、、、、、、、、、及。In some embodiments, M¹ -L¹ -T¹ and M² -L² -T² are each selected from 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 and .

如本文所述，L³係視情況經取代之C₁-C₆脂族基。在一些實施例中，L³係視情況經取代之C₁-C₆伸烷基。在一些實施例中，L³係-(CH₂)_1-6-。在一些實施例中，L³係–(CH₂)₃-或–(CH₂)₄-。在一些實施例中，L³係-(CH₂)₃-。在一些實施例中，L³係-(CH₂)₄-。As described herein,^L is an optionally substituted_C1 -_C6 aliphatic group. In some embodiments,^L is an optionally substituted_C1 -_C6 alkylene group. In some embodiments,^L is -(_CH2 )_1-6- . In some embodiments,^L is -(_CH2 )_3- or -(_CH2 )_4- . In some embodiments,^L is -(_CH2 )_3- . In some embodiments,^L is -(_CH2 )_4- .

如本文所述，G¹為-OH或-N(R^a)₂。在一些實施例中，G¹為-OH。在一些實施例中，G¹為-N(R^a)₂。在一些實施例中，G¹係-N(H)(R^a)。在一些實施例中，G¹係-N(C₁-C₆脂族基)(H)。在一些實施例中，G¹係-N(C₁-C₆脂族基)₂。在一些實施例中，G¹係-N(CH₃)₂。As described herein,^G is_-OH or -N(^Ra ). In some embodiments, G^is -OH. In some embodiments,^G is_-N (^Ra ). In some embodiments,^G is -N(H)(^Ra ). In some embodiments,^G is -N(_C1 -_C6 aliphatic)(H). In some embodiments,^G is -N(_C1 -_C6 aliphatic)₂ . In some embodiments,^G is -N(_CH3 )₂ .

在一些實施例中，-L³-G¹為-(CH₂)₃-OH或-(CH₂)₄-OH。在一些實施例中，-L³-G¹為-(CH₂)₃-N(CH₃)₂或-(CH₂)₄- N(CH₃)₂。In some embodiments, -L³ -G¹ is -(CH₂ )₃ -OH or -(CH₂ )₄ -OH. In some embodiments, -L³ -G¹ is -(CH₂ )₃ -N(CH₃ )₂ or -(CH₂ )₄ -N(CH₃ )₂ .

在一些實施例中，式I化合物由式II-1表示：II-1 或其醫藥學上可接受之鹽，其中G¹、L³、L¹、L²、p2及p3單獨及組合如本文之類別及子類中所定義，且其中*表示立體純碳原子。在式II-1之一些實施例中，用*指示之碳原子中之一者為(R)且另一者為(S)。在一些實施例中，用*指示之兩個碳原子均為(R)。在一些實施例中，用*指示之兩個碳原子均為(S)。In some embodiments, the compound of Formula I is represented by Formula II-1: II-1 or a pharmaceutically acceptable salt thereof, wherein G¹ , L³ , L¹ , L² , p2 , and p3 are as defined in classes and subclasses herein, individually and in combination, and wherein * represents a stereopure carbon atom. In some embodiments of Formula II-1, one of the carbon atoms indicated by * is (R ) and the other is (S ). In some embodiments, both carbon atoms indicated by * are (R ). In some embodiments, both carbon atoms indicated by * are (S ).

在一些實施例中，式I化合物由式II-2表示：II-2 或其醫藥學上可接受之鹽，其中G¹、L³、L¹及L²單獨及組合如本文之類別及子類中所定義，且其中*表示立體純碳原子。在式II-2之一些實施例中，用*指示之碳原子中之一者為(R)且另一者為(S)。在一些實施例中，用*指示之兩個碳原子均為(R)。在一些實施例中，用*指示之兩個碳原子均為(S)。In some embodiments, the compound of Formula I is represented by Formula II-2: II-2 or a pharmaceutically acceptable salt thereof, wherein G¹ , L³ , L¹ , and L² are as defined in classes and subclasses herein, individually and in combination, and wherein * represents a stereopure carbon atom. In some embodiments of Formula II-2, one of the carbon atoms indicated by * is (R ) and the other is (S ). In some embodiments, both carbon atoms indicated by * are (R ). In some embodiments, both carbon atoms indicated by * are (S ).

在一些實施例中，式I化合物由式II-3表示：II-3 或其醫藥學上可接受之鹽，其中L³、L¹、L²、p2及p3單獨及組合如本文之類別及子類中所定義，且其中*表示立體純碳原子。在式II-3之一些實施例中，用*指示之碳原子中之一者為(R)且另一者為(S)。在一些實施例中，用*指示之兩個碳原子均為(R)。在一些實施例中，用*指示之兩個碳原子均為(S)。In some embodiments, the compound of Formula I is represented by Formula II-3: II-3 or a pharmaceutically acceptable salt thereof, wherein L³ , L¹ , L² , p2 , and p3 are as defined in classes and subclasses herein, individually and in combination, and wherein * represents a stereopure carbon atom. In some embodiments of Formula II-3, one of the carbon atoms indicated by * is (R ) and the other is (S ). In some embodiments, both carbon atoms indicated by * are (R ). In some embodiments, both carbon atoms indicated by * are (S ).

在一些實施例中，式I化合物由式II-4表示：II-4 或其醫藥學上可接受之鹽，其中L³如本文之類別及子類中所定義，且其中*表示立體純碳原子。在式II-4之一些實施例中，用*指示之碳原子中之一者為(R)且另一者為(S)。在一些實施例中，用*指示之兩個碳原子均為(R)。在一些實施例中，用*指示之兩個碳原子均為(S)。In some embodiments, the compound of Formula I is represented by Formula II-4: II-4 or a pharmaceutically acceptable salt thereof, wherein^L is as defined in classes and subclasses herein, and wherein * represents a stereopure carbon atom. In some embodiments of Formula II-4, one of the carbon atoms indicated by * is (R ) and the other is (S ). In some embodiments, both carbon atoms indicated by * are (R ). In some embodiments, both carbon atoms indicated by * are (S ).

在一些實施例中，式I化合物選自表1： 表1In some embodiments, the compound of formula I is selected from Table 1: Table 1

在一些實施例中，所提供化合物係以鹽形式(例如，醫藥學上可接受之鹽形式)提供及/或使用。除非另有指示，否則提及本文所提供之化合物應理解為包括提及其鹽。In some embodiments, provided compounds are provided and/or used in salt form (e.g., pharmaceutically acceptable salt form). Unless otherwise indicated, reference to a compound provided herein should be understood to include reference to its salt.

如本文所述，在一些實施例中，式I化合物之特徵可在於具有約15,000與30,000之間的維納路徑(Wiener path)及-15與0之間的MMFF94能量。在一些實施例中，式I化合物具有約15,000與30,000之間的維納路徑。維納路徑(亦即維納指數)為分子之拓撲指數，其定義為化學圖中表示分子中之非氫原子的所有頂點對之間的最短路徑之長度之總和。在一些實施例中，式I化合物之MMFF94能量介於約-15與0之間。MMFF94能量係指Merck分子力場且為某些氫鍵結複合物提供構形能量。參見T. Halgren,J. Comput. Chem., 1999年5月; 20(7):730-748。As described herein, in some embodiments, compounds of Formula I can be characterized by having a Wiener path between about 15,000 and 30,000 and an MMFF94 energy between -15 and 0. In some embodiments, compounds of Formula I have a Wiener path between about 15,000 and 30,000. The Wiener path (also known as the Wiener index) is a topological index of a molecule, defined as the sum of the lengths of the shortest paths between all pairs of vertices in a chemical graph representing non-hydrogen atoms in the molecule. In some embodiments, compounds of Formula I have an MMFF94 energy between about -15 and 0. The MMFF94 energy refers to the Merck molecular force field and provides conformational energies for certain hydrogen-bonded complexes.See T. Halgren,J. Comput. Chem. , May 1999; 20(7):730-748.

在一些實施例中，本文所述之醫藥組合物(例如，為或包含脂質奈米粒子之組合物)包含相對於該組合物中脂質之總量約20 mol%至約70 mol%之陽離子脂質(例如，式I化合物或式II-1至II-4中任一者之化合物)。在一些實施例中，醫藥組合物包含相對於該組合物中脂質之總量約40至約50 mol%之陽離子脂質。在一些實施例中，醫藥組合物包含相對於該組合物中脂質之總量約30 mol%至約60 mol%之陽離子脂質。In some embodiments, the pharmaceutical compositions described herein (e.g., compositions that are or include lipid nanoparticles) comprise from about 20 mol% to about 70 mol% of a cationic lipid (e.g., a compound of Formula I or a compound of any one of Formulas II-1 to II-4), relative to the total amount of lipids in the composition. In some embodiments, the pharmaceutical compositions comprise from about 40 to about 50 mol% of a cationic lipid, relative to the total amount of lipids in the composition. In some embodiments, the pharmaceutical compositions comprise from about 30 mol% to about 60 mol% of a cationic lipid, relative to the total amount of lipids in the composition.

脂質奈米粒子在一些實施例中，本文所述之式I化合物與核酸複合以形成核酸粒子。在一些實施例中，核酸粒子係脂質奈米粒子。在一些實施例中，脂質奈米粒子係陽離子脂質奈米粒子，其包含一或多種陽離子脂質(例如，本文所述之陽離子脂質)、核酸(例如，RNA及/或DNA)，及一種或多種額外脂質(例如，聚合物偶聯脂質、類固醇、陰離子兩親物及/或中性脂質) 脂質奈米粒子(LNP)已證實可用於將核酸負荷遞送至所關注組織。LNP用於例如治療COVID-19之某些商業疫苗中。Lipid Nanoparticles In some embodiments, a compound of Formula I described herein is complexed with a nucleic acid to form a nucleic acid particle. In some embodiments, the nucleic acid particle is a lipid nanoparticle. In some embodiments, the lipid nanoparticle is a cationic lipid nanoparticle, which comprises one or more cationic lipids (e.g. , cationic lipids described herein), a nucleic acid (e.g., RNA and/or DNA), and one or more additional lipids (e.g., polymer-conjugated lipids, steroids, anionic amphiphiles, and/or neutral lipids). Lipid nanoparticles (LNPs) have been shown to be useful for delivering nucleic acid cargo to tissues of interest. LNPs are used, for example, in certain commercial vaccines for the treatment of COVID-19.

在一些實施例中，本揭示案提供一種醫藥組合物，其包含本文所述的陽離子或可離子化脂質化合物(例如，式I、II-1、II-2、II-3或II-4化合物)及核酸。在一些實施例中，陽離子或可離子化脂質化合物(例如式I、II-1、II-2、II-3或II-4化合物)及核酸係呈脂質奈米粒子(LNP)形式。In some embodiments, the present disclosure provides a pharmaceutical composition comprising a cationic or ionizable lipid compound described herein (e.g., a compound of Formula I, II-1, II-2, II-3, or II-4) and a nucleic acid. In some embodiments, the cationic or ionizable lipid compound (e.g., a compound of Formula I, II-1, II-2, II-3, or II-4) and the nucleic acid are in the form of lipid nanoparticles (LNPs).

本文所述之核酸粒子(例如，核糖核酸粒子或去氧核糖核酸粒子)之特徵可在於「N/P比」，其係陽離子聚合物中之陽離子(氮)基團(N/P中之「N」)對RNA中之陰離子(磷酸根)基團(N/P中之「P」)的莫耳比。應理解，陽離子基團係呈陽離子形式(例如，N⁺)之基團，或可離子化變成陽離子之基團。在N/P比中使用單個數字(例如，約5之N/P比)意欲指該數字超過1，例如，約4之N/P比意欲意指約4:1。在一些實施例中，本文所述之醫藥組合物(例如，在核酸粒子，諸如脂質奈米粒子中)之N/P比為約3至約12。在一些實施例中，N/P比為3、4、5、6、7、8、9、10、11或12。在一些實施例中，N/P比為約4至約8。在一些實施例中，N/P比為4。在一些實施例中，N/P比為5。在一些實施例中，在一些實施例中，N/P比為6。在一些實施例中，N/P比為7。在一些實施例中，N/P比為8。The nucleic acid particles described herein (e.g., RNA particles or DNA particles) can be characterized by an "N/P ratio," which is the molar ratio of cationic (nitrogen) groups in the cationic polymer (the "N" in N/P) to anionic (phosphate) groups in the RNA (the "P" in N/P). It should be understood that cationic groups are groups that are in a cationic form (e.g., N⁺ ) or that can be ionized to become cationic. The use of a single number in the N/P ratio (e.g., an N/P ratio of about 5) is intended to mean that the number is greater than 1; for example, an N/P ratio of about 4 is intended to mean about 4:1. In some embodiments, the pharmaceutical compositions described herein (e.g., in nucleic acid particles, such as lipid nanoparticles) have an N/P ratio of about 3 to about 12. In some embodiments, the N/P ratio is 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12. In some embodiments, the N/P ratio is about 4 to about 8. In some embodiments, the N/P ratio is 4. In some embodiments, the N/P ratio is 5. In some embodiments, in some embodiments, the N/P ratio is 6. In some embodiments, the N/P ratio is 7. In some embodiments, the N/P ratio is 8.

在一些實施例中，包含本文所述的陽離子或可離子化脂質化合物(例如，式I、II-1、II-2、II-3或II-4化合物)及核酸之醫藥組合物進一步包含額外脂質。在一些實施例中，包含本文所述之陽離子或可離子化脂質化合物(例如式I、II-1、II-2、II-3或II-4之化合物)及核酸之醫藥組合物進一步包含中性脂質、聚合物偶聯脂質、類固醇或陰離子兩親物中之一或多者。In some embodiments, the pharmaceutical composition comprising a cationic or ionizable lipid compound described herein (e.g., a compound of Formula I, II-1, II-2, II-3, or II-4) and a nucleic acid further comprises an additional lipid. In some embodiments, the pharmaceutical composition comprising a cationic or ionizable lipid compound described herein (e.g., a compound of Formula I, II-1, II-2, II-3, or II-4) and a nucleic acid further comprises one or more of a neutral lipid, a polymer-conjugated lipid, a steroid, or an anionic amphiphile.

在一些實施例中，包含本文所述的陽離子或可離子化脂質化合物(例如，式I、II-1、II-2、II-3或II-4化合物)及核酸之醫藥組合物進一步包含中性脂質、聚合物偶聯脂質及類固醇。在一些實施例中，包含本文所述的陽離子或可離子化脂質化合物(例如，式I、II-1、II-2、II-3或II-4化合物)及核酸之醫藥組合物進一步包含中性脂質、陰離子兩親物及類固醇。在一些實施例中，包含本文所述之陽離子或可離子化脂質化合物(例如式I、II-1、II-2、II-3或II-4之化合物)及核酸之醫藥組合物進一步包含中性脂質、陰離子兩親物及類固醇，且其中該藥物組合物不包含聚合物偶聯脂質。In some embodiments, the pharmaceutical composition comprising a cationic or ionizable lipid compound described herein (e.g., a compound of Formula I, II-1, II-2, II-3, or II-4) and a nucleic acid further comprises a neutral lipid, a polymer-conjugated lipid, and a steroid. In some embodiments, the pharmaceutical composition comprising a cationic or ionizable lipid compound described herein (e.g., a compound of Formula I, II-1, II-2, II-3, or II-4) and a nucleic acid further comprises a neutral lipid, an anionic amphiphile, and a steroid. In some embodiments, the pharmaceutical composition comprising a cationic or ionizable lipid compound described herein (e.g., a compound of Formula I, II-1, II-2, II-3, or II-4) and a nucleic acid further comprises a neutral lipid, an anionic amphiphile, and a steroid, wherein the pharmaceutical composition does not comprise a polymer-conjugated lipid.

(i) 中性脂質 如本文所述，在一些實施例中，本文所述之醫藥組合物進一步包含中性脂質。在一些實施例中，中性脂質係磷脂。在一些實施例中，中性脂質係或包含1,2-二硬脂醯基-sn-甘油-3-磷酸膽鹼(DSPC)、1,2-二棕櫚醯基-sn-甘油-3-磷酸膽鹼(DPPC)、1,2-二肉豆蔻醯基-sn-甘油-3-磷酸膽鹼(DMPC)、1-棕櫚醯基-2-油醯基-sn-甘油-3-磷酸膽鹼(POPC)、1,2-二油醯基-sn-甘油-3-磷酸膽鹼(DOPC)、磷脂醯乙醇胺(諸如1,2-二油醯基-sn-甘油-3-磷酸乙醇胺(DOPE))、鞘磷脂(SM)、1,2‑二醯基甘油-3-O-4'-(N,N,N-三甲基)-高絲胺酸 (DGTS)、神經醯胺、膽固醇、類固醇(諸如固醇及其衍生物)。(i) Neutral lipids As described herein, in some embodiments, the pharmaceutical compositions described herein further comprise a neutral lipid. In some embodiments, the neutral lipid is a phospholipid. In some embodiments, the neutral lipid may comprise 1,2-distearoyl-sn -glycero-3-phosphocholine (DSPC), 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), 1,2-dioleoyl-sn-glycero- 3-phosphocholine (DOPC), phosphoethanolamines (such as 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE)), sphingomyelin (SM), 1,2-diacylglycero-3-O-4'-(N,N,N-trimethyl)-homoserine (DGTS), ceramide, cholesterol, steroids (such as sterols and their derivatives).

在一些實施例中，中性脂質係或包含磷脂醯膽鹼、磷脂醯乙醇胺、磷脂醯甘油、磷脂酸、磷脂醯絲胺酸或鞘磷脂。在一些實施例中，中性脂質係或包含二醯基磷脂醯膽鹼，諸如二硬脂醯基磷脂醯膽鹼(DSPC)、二油醯基磷脂醯膽鹼(DOPC)、二肉豆蔻醯基磷脂醯膽鹼(DMPC)、雙十五醯基磷脂醯膽鹼、二月桂醯基磷脂醯膽鹼、二棕櫚醯基磷脂醯膽鹼(DPPC)、二花生醯基磷脂醯膽鹼(DAPC)、二山崳醯基磷脂醯膽鹼(DBPC)、雙二十三醯基磷脂醯膽鹼(DTPC)、雙二十四醯基磷脂醯膽鹼(DLPC)、棕櫚醯基油醯基-磷脂醯膽鹼(POPC)、1,2-二-O-十八烯基-sn-甘油-3-磷酸膽鹼(18:0二醚PC)、1-油醯基-2-膽固醇基半琥珀醯基-sn-甘油-3-磷酸膽鹼(OChemsPC)、1-十六基-sn-甘油-3-磷酸膽鹼(C16 Lyso PC)及磷脂醯乙醇胺，包括例如二醯基磷脂醯乙醇胺，諸如二油醯基磷脂醯乙醇胺(DOPE)、二硬脂醯基-磷脂醯乙醇胺(DSPE)、二棕櫚醯基-磷脂醯乙醇胺(DPPE)、二肉豆蔻醯基-磷脂醯乙醇胺(DMPE)、二月桂醯基-磷脂醯乙醇胺(DLPE)、二植醯基-磷脂醯乙醇胺(DPyPE)、1,2-二-(9Z-十八碳烯醯基)-sn-甘油-3-磷酸膽鹼(DOPG)、1,2-二棕櫚醯基-sn-甘油-3-磷酸-(1′-rac-甘油) (DPPG)、1-棕櫚醯基-2-油醯基-sn-甘油-3-磷酸乙醇胺(POPE)、N-棕櫚醯基-D-赤-鞘胺醯基磷酸膽鹼(SM)。在一些實施例中，中性脂質選自由以下組成之群：DSPC、DOPC、DMPC、DPPC、POPC、DOPE、DOPG、DPPG、POPE、DPPE、DMPE、DSPE及SM。在一些實施例中，中性脂質選自由以下組成之群：DSPC、DPPC、DMPC、DOPC、POPC、DOPE及SM。在一些實施例中，中性脂質係DSPC。In some embodiments, the neutral lipid is or comprises phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol, phosphatidic acid, phosphatidylserine, or sphingomyelin. In some embodiments, the neutral lipid is or comprises a diacylphospholipid acylcholine, such as distearylphospholipid acylcholine (DSPC), dioleoylphospholipid acylcholine (DOPC), dimyristoylphospholipid acylcholine (DMPC), dipentadecyloylphospholipid acylcholine, dilaurylphospholipid acylcholine, dipalmitoylphospholipid acylcholine (DPPC), diarachidylphospholipid acylcholine (DAPC), dibehenylphospholipid acylcholine (DBPC), bis(dipentadecyloylphospholipid acylcholine), dilaurylphospholipid acylcholine, dipalmitoylphospholipid acylcholine (DPPC), diarachidylphospholipid acylcholine (DAPC), dibehenylphospholipid acylcholine (DBPC), dioctylphospholipid acylcholine (D ... Tridecylphosphatidylcholine (DTPC), di-tetracosylphosphatidylcholine (DLPC), palmitoyloleylphosphatidylcholine (POPC), 1,2-di-O-octadecenyl-sn-glycero-3-phosphocholine (18:0 diether PC), 1-oleyl-2-cholestyryl hemisuccinyl-sn-glycero-3-phosphocholine (OChemsPC), 1-hexadecyl-sn-glycero-3-phosphocholine (C16 Lyso PC) and phosphatidylethanolamines, including, for example, diacylphosphatidylethanolamines, such as dioleylphosphatidylethanolamine (DOPE), distearyl-phosphatidylethanolamine (DSPE), dimalmitoyl-phosphatidylethanolamine (DPPE), dimyristoyl-phosphatidylethanolamine (DMPE), dilauryl-phosphatidylethanolamine (DLPE), diphytyl-phosphatidylethanolamine (DPyPE), 1,2-bis-(9Z-octadecenyl)-sn-glycero-3-phosphocholine (DOPG), 1,2-dipalmitoyl-sn-glycero-3-phospho-(1′-rac-glycerol) (DPPG), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine (POPE), N-palmitoyl-D-erythro-sphingamidoylphosphocholine (SM). In some embodiments, the neutral lipid is selected from the group consisting of DSPC, DOPC, DMPC, DPPC, POPC, DOPE, DOPG, DPPG, POPE, DPPE, DMPE, DSPE, and SM. In some embodiments, the neutral lipid is selected from the group consisting of DSPC, DPPC, DMPC, DOPC, POPC, DOPE, and SM. In some embodiments, the neutral lipid is DSPC.

中性脂質可為合成或天然衍生的。適用於醫藥組合物中之其他中性脂質描述於WO2021/026358、WO 2017/075531及WO 2018/081480中，該等專利中每一者之全部內容之全文皆以引用方式併入本文中。Neutral lipids can be synthetic or naturally derived. Other neutral lipids suitable for use in pharmaceutical compositions are described in WO 2021/026358, WO 2017/075531, and WO 2018/081480, the entire contents of each of which are incorporated herein by reference.

在一些實施例中，醫藥組合物包含約1 mol%至約40 mol%之中性脂質(相對於該醫藥組合物中之總脂質)。在一些實施例中，醫藥組合物包含約2 mol%至約25 mol%之中性脂質(相對於該醫藥組合物中之總脂質)。在一些實施例中，醫藥組合物包含約5 mol%至約15 mol%之中性脂質(相對於該醫藥組合物中之總脂質)。在一些實施例中，醫藥組合物包含約8 mol%至約12 mol%之中性脂質(相對於該醫藥組合物中之總脂質)。在一些實施例中，醫藥組合物包含約10 mol%之中性脂質(相對於該醫藥組合物中之總脂質)。In some embodiments, the pharmaceutical composition comprises about 1 mol% to about 40 mol% neutral lipids (relative to the total lipids in the pharmaceutical composition). In some embodiments, the pharmaceutical composition comprises about 2 mol% to about 25 mol% neutral lipids (relative to the total lipids in the pharmaceutical composition). In some embodiments, the pharmaceutical composition comprises about 5 mol% to about 15 mol% neutral lipids (relative to the total lipids in the pharmaceutical composition). In some embodiments, the pharmaceutical composition comprises about 8 mol% to about 12 mol% neutral lipids (relative to the total lipids in the pharmaceutical composition). In some embodiments, the pharmaceutical composition comprises about 10 mol% neutral lipids (relative to the total lipids in the pharmaceutical composition).

在一些實施例中，醫藥組合物包含約1 mol%至約40 mol%之磷脂。在一些實施例中，醫藥組合物包含約2 mol%至約25 mol%之磷脂。在一些實施例中，醫藥組合物包含約5 mol%至約15 mol%之磷脂。在一些實施例中，醫藥組合物包含約8 mol%至約12 mol%之磷脂。在一些實施例中，醫藥組合物包含約10 mol%之磷脂。在一些實施例中，醫藥組合物包含約5 mol%至約20 mol%之DSPC。在一些實施例中，醫藥組合物包含約8 mol%至約12 mol%之DSPC。在一些實施例中，醫藥組合物包含約10 mol%之DSPC。In some embodiments, the pharmaceutical composition comprises about 1 mol% to about 40 mol% phospholipids. In some embodiments, the pharmaceutical composition comprises about 2 mol% to about 25 mol% phospholipids. In some embodiments, the pharmaceutical composition comprises about 5 mol% to about 15 mol% phospholipids. In some embodiments, the pharmaceutical composition comprises about 8 mol% to about 12 mol% phospholipids. In some embodiments, the pharmaceutical composition comprises about 10 mol% phospholipids. In some embodiments, the pharmaceutical composition comprises about 5 mol% to about 20 mol% DSPC. In some embodiments, the pharmaceutical composition comprises about 8 mol% to about 12 mol% DSPC. In some embodiments, the pharmaceutical composition comprises about 10 mol% DSPC.

(ii) 聚合物偶聯脂質 如本文所述，在一些實施例中，本文所述之醫藥組合物進一步包含聚合物偶聯脂質。在一些實施例中，聚合物偶聯脂質係偶聯至聚乙二醇之脂質(PEG-脂質)。在一些實施例中，PEG脂質選自聚乙二醇化二醯基甘油(PEG-DAG) (諸如l-(單甲氧基-聚乙二醇)-2,3-二肉豆蔻醯甘油(PEG-DMG)，例如1,2-二肉豆蔻醯基-rac-甘油-3-甲氧基聚乙二醇-2000 (PEG2000-DMG))、聚乙二醇化磷脂醯乙醇胺(PEG-PE)、PEG二醯基甘油琥珀酸酯(PEG-S-DAG) (諸如4-O-(2',3'-二(十四醯氧基)丙基-1-O-(ω-甲氧基(聚乙氧基)乙基)丁二酸酯(PEG-S-DMG))、1,2-二硬脂醯基-sn-甘油-3-磷酸乙醇胺-N-[胺基(聚乙二醇)-2000] (DSPE-PEG2000胺)、聚乙二醇化神經醯胺(PEG-cer)或PEG二烷氧基丙基胺基甲酸酯(諸如ω-甲氧基(聚乙氧基)乙基-N-(2,3-二(十四烷氧基)丙基)胺基甲酸酯、2,3-二(十四烷氧基)丙基-1-N-(ω-甲氧基(聚乙氧基)乙基)胺基甲酸酯)及N-棕櫚醯基-鞘胺醇-1-{琥珀醯基[甲氧基(聚乙二醇)2000]} (C16-PEG2000神經醯胺或C16Cer-PEG2K)。(ii) Polymer-Conjugated LipidsAs described herein, in some embodiments, the pharmaceutical compositions described herein further comprise a polymer-conjugated lipid. In some embodiments, the polymer-conjugated lipid is a lipid conjugated to polyethylene glycol (PEG-lipid). In some embodiments, the PEG lipid is selected from PEGylated diacylglycerol (PEG-DAG) (e.g., 1-(monomethoxy-polyethylene glycol)-2,3-dimyristoylglycerol (PEG-DMG), e.g., 1,2-dimyristoyl-rac-glycero-3-methoxypolyethylene glycol-2000 (PEG2000-DMG)), PEGylated phosphatidylethanolamine (PEG-PE), PEG diacylglycerol succinate (PEG-S-DAG) (e.g., 4-O-(2',3'-bis(tetradecanoyloxy)propyl)-1-O-(ω-methoxy(polyethoxy)ethyl)succinate (PEG-S-DMG)), 1,2-distearyl-sn-glycero-3-phosphoethanolamine-N-[amino(polyethylene glycol)-2000] (DSPE-PEG2000 amine), PEGylated ceramide (PEG-cer) or PEG dialkoxypropyl carbamate (such as ω-methoxy(polyethoxy)ethyl-N-(2,3-di(tetradecyloxy)propyl) carbamate, 2,3-di(tetradecyloxy)propyl-1-N-(ω-methoxy(polyethoxy)ethyl) carbamate) and N-palmitoyl-sphingosine-1-{succinyl[methoxy(polyethylene glycol)2000]} (C16-PEG2000 ceramide or C16Cer-PEG2K).

在一些實施例中，PEG-脂質係PEG2000-DMG：In some embodiments, the PEG-lipid is PEG2000-DMG:

在一些實施例中，PEG-脂質係DMG-PEG。In some embodiments, the PEG-lipid is DMG-PEG.

在一些實施例中，PEG-脂質提供於WO2021/026358、WO 2017/075531或WO 2018/081480中，該等專利中每一者之全文皆以引用方式併入。In some embodiments, the PEG-lipid is provided in WO 2021/026358, WO 2017/075531, or WO 2018/081480, each of which is incorporated by reference in its entirety.

在一些實施例中，聚合物偶聯脂質係C16 PEG2000：In some embodiments, the polymer-coupled lipid is C16 PEG2000:

在一些實施例中，PEG-脂質具有以下結構：其中上式中之n係30至60，例如約50。在一個實施例中，PEG偶聯脂質(聚乙二醇化脂質)係PEG₂₀₀₀-C-DMA，其較佳地係指3-N-[(ω-甲氧基聚(乙二醇)2000)胺甲醯基]-1,2-二肉豆蔻氧基-丙胺(MPEG-(2 kDa)-C-DMA)或甲氧基-聚乙二醇-2,3-雙(十四基氧基)丙基胺基甲酸酯(2000)。In some embodiments, the PEG-lipid has the following structure: wherein n in the above formula is 30 to 60, for example, about 50. In one embodiment, the PEG-conjugated lipid (PEGylated lipid) is PEG₂₀₀₀ -C-DMA, preferably 3-N-[(ω-methoxypoly(ethylene glycol) 2000)aminoformyl]-1,2-dimyristyloxy-propylamine (MPEG-(2 kDa)-C-DMA) or methoxy-poly(ethylene glycol)-2,3-bis(tetradecyloxy)propylcarbamate (2000).

在一些實施例中，PEG-脂質選自PEG-DAG、PEG-PE、PEG-S-DAG、PEG2000-DMG、PEG-S-DMG、PEG-cer、PEG二烷氧基丙基胺基甲酸酯(例如，ω-甲氧基(聚乙氧基)乙基-N-(2,3-二(十四烷氧基)丙基)胺基甲酸酯或2,3-二(十四烷氧基)丙基-N-(ω-甲氧基(聚乙氧基)乙基)胺基甲酸酯)及其組合。在一些實施例中，PEG-脂質係PEG2000-DMG。在一些實施例中，PEG-脂質係PEG-DAG。在一些實施例中，PEG-脂質係PEG-PE。在一些實施例中，PEG-脂質係PEG-S-DAG。在一些實施例中，PEG-脂質係PEG-cer。在一些實施例中，PEG-脂質係PEG二烷氧基丙基胺基甲酸酯。In some embodiments, the PEG-lipid is selected from PEG-DAG, PEG-PE, PEG-S-DAG, PEG2000-DMG, PEG-S-DMG, PEG-cer, PEG dialkoxypropylcarbamate (e.g., ω-methoxy(polyethoxy)ethyl-N-(2,3-di-tetradecyloxy)propyl)carbamate or 2,3-di-tetradecyloxypropyl-N-(ω-methoxy(polyethoxy)ethyl)carbamate), and combinations thereof. In some embodiments, the PEG-lipid is PEG2000-DMG. In some embodiments, the PEG-lipid is PEG-DAG. In some embodiments, the PEG-lipid is PEG-PE. In some embodiments, the PEG-lipid is PEG-S-DAG. In some embodiments, the PEG-lipid is PEG-cer. In some embodiments, the PEG-lipid is PEG dialkoxypropylcarbamate.

在一些實施例中，為PEG-脂質之一部分之PEG基團在包含一或多種PEG-脂質分子之組合物中平均具有約2000 g/mol之重量平均分子量(M_w)。In some embodiments, the PEG group that is part of the PEG-lipid has, on average, a weight average molecular weight (_Mw ) of about 2000 g/mol in a composition comprising one or more PEG-lipid molecules.

在一些實施例中，聚合物偶聯脂質係聚肌胺酸偶聯脂質，在本文中亦稱為肌胺酸化脂質或pSar-脂質。術語「肌胺酸化脂質」係指包含脂質部分及聚肌胺酸(聚(N-甲基甘胺酸))部分之分子。In some embodiments, the polymer-conjugated lipid is a poly(sarcosine)-conjugated lipid, also referred to herein as a sarcosinated lipid or pSar-lipid. The term "sarcosinated lipid" refers to a molecule comprising a lipid portion and a poly(sarcosine) (poly(N-methylglycine)) portion.

在一些實施例中，聚合物偶聯脂質係聚噁唑啉(POX)偶聯脂質及/或聚噁嗪(POZ)偶聯脂質，在本文中亦稱為POX及/或POZ聚合物及一或多條疏水鏈之偶聯物或稱為噁唑啉化及/或噁嗪化脂質或POX-及/或POZ-脂質。術語「噁唑啉化脂質」或「POX-脂質」係指包含脂質部分及聚噁唑啉部分之分子。術語「噁嗪化脂質」或「POZ-脂質」係指包含脂質部分及聚噁嗪部分之分子。術語「噁唑啉化/噁嗪化脂質」或「POX/POZ-脂質」或「POXZ-脂質」係指包含脂質部分以及聚噁唑啉及聚噁嗪之共聚物之部分的分子。In some embodiments, the polymer-conjugated lipid is a polyoxazoline (POX)-conjugated lipid and/or a polyoxazine (POZ)-conjugated lipid, also referred to herein as a conjugate of a POX and/or POZ polymer and one or more hydrophobic chains, or as an oxazolinated and/or oxazinated lipid or a POX- and/or POZ-lipid. The term "oxazolinated lipid" or "POX-lipid" refers to a molecule comprising a lipid portion and a polyoxazoline portion. The term "oxazinated lipid" or "POZ-lipid" refers to a molecule comprising a lipid portion and a polyoxazine portion. The term "oxazolinated/oxazinated lipid" or "POX/POZ-lipid" or "POXZ-lipid" refers to a molecule comprising a lipid portion and a copolymer of polyoxazoline and polyoxazine.

在一些實施例中，本文所述之LNP可包含肌胺酸化脂質。在一些實施例中，本文所述之核酸組合物(例如DNA或RNA組合物，尤其mRNA組合物)包含肌胺酸化脂質且實質上不含聚乙二醇化脂質(或不含聚乙二醇化脂質)。In some embodiments, the LNPs described herein may comprise sarcosinate lipids. In some embodiments, the nucleic acid compositions described herein (e.g., DNA or RNA compositions, particularly mRNA compositions) comprise sarcosinate lipids and are substantially free of PEGylated lipids (or free of PEGylated lipids).

在一些實施例中，本文所述之核酸組合物(例如DNA或RNA組合物)包含如本文所述之陽離子/陽離子可離子化脂質及肌胺酸化脂質(pSAR偶聯脂質)。在一些實施例中，本文所述之核酸組合物(例如DNA或RNA組合物，尤其mRNA組合物)可進一步包含中性脂質(例如，磷脂、膽固醇或其衍生物)或中性脂質(例如，磷脂及膽固醇或其衍生物)之組合。在一些實施例中，本文所述之核酸組合物(例如DNA或RNA組合物，尤其mRNA組合物)包含如本文所述之陽離子/陽離子可離子化脂質、肌胺酸化脂質、中性脂質(例如，磷脂)及膽固醇或其衍生物。在一些實施例中，磷脂係DSPC。In some embodiments, the nucleic acid compositions described herein (e.g., DNA or RNA compositions) comprise cations/cationically ionizable lipids and sarcosinate lipids (pSAR-coupled lipids) as described herein. In some embodiments, the nucleic acid compositions described herein (e.g., DNA or RNA compositions, especially mRNA compositions) may further comprise neutral lipids (e.g., phospholipids, cholesterol, or derivatives thereof) or a combination of neutral lipids (e.g., phospholipids and cholesterol, or derivatives thereof). In some embodiments, the nucleic acid compositions described herein (e.g., DNA or RNA compositions, especially mRNA compositions) comprise cations/cationically ionizable lipids, sarcosinate lipids, neutral lipids (e.g., phospholipids), and cholesterol, or derivatives thereof, as described herein. In some embodiments, the phospholipid is DSPC.

在包含肌胺酸化脂質之本文所述之核酸組合物(例如DNA或RNA組合物，尤其mRNA組合物)之一些實施例中，該等組合物實質上不含聚乙二醇化脂質(或不含聚乙二醇化脂質)。In some embodiments of the nucleic acid compositions described herein (e.g., DNA or RNA compositions, particularly mRNA compositions) that comprise sarcosinate lipids, the compositions are substantially free of PEGylated lipids (or contain no PEGylated lipids).

在一些實施例中，肌胺酸化脂質包含2至200個肌胺酸單元，例如5至100個肌胺酸單元、10至50個肌胺酸單元、15至40個肌胺酸單元，例如約23個肌胺酸單元。In some embodiments, the sarcosinate lipid comprises 2 to 200 sarcosine units, e.g., 5 to 100 sarcosine units, 10 to 50 sarcosine units, 15 to 40 sarcosine units, e.g., about 23 sarcosine units.

在一些實施例中，肌胺酸化脂質包含以下通式(XVII)之結構：XVII 其中s係肌胺酸單元之數量。In some embodiments, the sarcosinated lipid comprises the following structure of Formula (XVII): XVII where s is the number of sarcosine units.

在一些實施例中，肌胺酸化脂質包含以下通式(XVIII)之結構：XVIII 其中R₂₁及R₂₂中之一者包含疏水基團且另一者係H、親水基團或視情況地包含靶向部分之官能基；且x係肌胺酸單元之數量。In some embodiments, the sarcosinated lipid comprises the following structure of Formula (XVIII): XVIII wherein one of R₂₁ and R₂₂ comprises a hydrophobic group and the other is H, a hydrophilic group, or optionally comprises a functional group of a targeting moiety; and x is the number of sarcosine units.

在一些實施例中，本文之LNP可包含噁唑啉化及/或噁嗪化脂質。在一些實施例中，本文所述之核酸組合物(例如DNA或RNA組合物，尤其mRNA組合物)包含噁唑啉化及/或噁嗪化脂質且實質上不含聚乙二醇化脂質(或不含聚乙二醇化脂質)。In some embodiments, the LNPs herein may comprise oxazolinylated and/or oxazinylated lipids. In some embodiments, the nucleic acid compositions described herein (e.g., DNA or RNA compositions, particularly mRNA compositions) comprise oxazolinylated and/or oxazinylated lipids and are substantially free of PEGylated lipids (or free of PEGylated lipids).

在一些實施例中，聚合物偶聯脂質包含2-(2-(2-胺基乙氧基)乙氧基)乙酸之單體。在一些實施例中，聚合物偶聯脂質包含2-(2-(2-胺基乙氧基)乙氧基)乙酸之單體。在一些實施例中，聚合物偶聯脂質之聚合物係或包含聚-2-(2-(2-胺基乙氧基)乙氧基)乙酸(pAEEA)或聚-2-(2-(2-甲基胺基乙氧基)乙氧基)乙酸(pMAEEA)或其衍生物，如本文所定義。In some embodiments, the polymer-conjugated lipid comprises a monomer of 2-(2-(2-aminoethoxy)ethoxy)acetic acid. In some embodiments, the polymer-conjugated lipid comprises a monomer of 2-(2-(2-aminoethoxy)ethoxy)acetic acid. In some embodiments, the polymer of the polymer-conjugated lipid comprises poly-2-(2-(2-aminoethoxy)ethoxy)acetic acid (pAEEA) or poly-2-(2-(2-methylaminoethoxy)ethoxy)acetic acid (pMAEEA), or a derivative thereof, as defined herein.

在一些實施例中，聚合物包含以下通式：其中X¹¹及X¹²一起係視情況經取代之醯胺、視情況經取代之硫代醯胺或酯；Y係-CH₂-、-(CH₂)₂-或-(CH₂)₃-；z係2至24；且n係1至100。In some embodiments, the polymer comprises the following formula: wherein X¹¹ and X¹² together are optionally substituted amide, optionally substituted thioamide, or ester; Y is -CH₂ -, -(CH₂ )₂ -, or -(CH₂ )₃ -; z is 2 to 24; and n is 1 to 100.

在一些實施例中，(i)當X¹¹係-C(O)-時，則X¹²係-NR¹⁰-；(ii)當X¹¹係-NR¹⁰-時，則X¹²係-C(O)-；(iii)當X¹¹係-C(S)-時，則X¹²係-NR¹-；(iv)當X¹¹係-NR¹-時，則X¹²係-C(S)-；(v)當X¹¹係-C(O)-時，則X¹²係-O-；或(vi)當X¹¹係-O-時，則X¹²係-C(O)-；其中R¹⁰係氫或C_1-8烷基。在一些實施例中，X¹¹係-C(O)-且X¹²係-NR¹⁰-，其中R¹⁰係氫或C_1-8烷基。在一些實施例中，X¹¹係-C(O)-且X¹²係-NR¹⁰-，其中R¹⁰係氫或甲基。在一些實施例中，X¹¹係-C(O)-且X¹²係-NR¹⁰-，其中R¹⁰係氫。在一些實施例中，Y係-CH₂-或-(CH₂)₂-。在一些實施例中，Y係-CH₂-。In some embodiments, (i) when X¹¹ is -C(O)-, then X¹² is -NR¹⁰ -; (ii) when X¹¹ is -NR¹⁰ -, then X¹² is -C(O)-; (iii) when X¹¹ is -C(S)-, then X¹² is -NR¹ -; (iv) when X¹¹ is -NR¹ -, then X¹² is -C(S)-; (v) when X¹¹ is -C(O)-, then X¹² is -O-; or (vi) when X¹¹ is -O-, then X¹² is -C(O)-; wherein R¹⁰ is hydrogen or C_1-8 alkyl. In some embodiments, X¹¹ is -C(O)- and X¹² is -NR¹⁰ -, wherein R¹⁰ is hydrogen or C_1-8 alkyl. In some embodiments, X¹¹ is -C(O)- and X¹² is -NR¹⁰ -, wherein R¹⁰ is hydrogen or methyl. In some embodiments, X¹¹ is -C(O)- and X¹² is -NR¹⁰ -, wherein R¹⁰ is hydrogen. In some embodiments, Y is -CH₂ - or -(CH₂ )₂ -. In some embodiments, Y is -CH₂ -.

在一些實施例中，聚合物包含以下通式：其中R¹⁰係氫或C1-8烷基；z係2至24；且n係1至100。在上式之一些實施例中，z係2至10。在一些實施例中，z係2至5。在一些實施例中，z係2。在上式之一些實施例中，R¹⁰係氫或甲基。在一些實施例中，R¹⁰係氫。In some embodiments, the polymer comprises the following formula: wherein R¹⁰ is hydrogen or C 1-8 alkyl; z is 2 to 24; and n is 1 to 100. In some embodiments of the above formula, z is 2 to 10. In some embodiments, z is 2 to 5. In some embodiments, z is 2. In some embodiments of the above formula, R¹⁰ is hydrogen or methyl. In some embodiments, R¹⁰ is hydrogen.

在一些實施例中，聚合物偶聯脂質包含以下結構之「n」個單體：In some embodiments, the polymer-conjugated lipid comprises "n" monomers of the following structure:

在上式之一些實施例中，n係5至50。在一些實施例中，n係5至25。在一些實施例中，n係7至14。在一些實施例中，n係10至25。在一些實施例中，n係14至17。在一些實施例中，n係8或14。In some embodiments of the above formula, n is 5 to 50. In some embodiments, n is 5 to 25. In some embodiments, n is 7 to 14. In some embodiments, n is 10 to 25. In some embodiments, n is 14 to 17. In some embodiments, n is 8 or 14.

在一些實施例中，聚合物偶聯脂質包含以下結構之單體：In some embodiments, the polymer-conjugated lipid comprises a monomer having the following structure:

在一些實施例中，聚合物偶聯脂質選自下表：DSPE-AEEA14-ACVE-AEEA14-ACVE-(AEEA)8-ACDMA-(AEEA)14-ACDMA-(AEEA)8-ACDMG-(AEEA)8-ACIn some embodiments, the polymer-conjugated lipid is selected from the following table: DSPE-AEEA14-AC VE-AEEA14-AC VE-(AEEA)8-AC DMA-(AEEA)14-AC DMA-(AEEA)8-AC DMG-(AEEA)8-AC

在一些實施例中，相對於LNP中之總脂質，聚合物偶聯脂質係約0.5 mol%至約5 mol%。在一些實施例中，LNP包含約1.0 mol%至約2.5 mol%之聚合物偶聯脂質。在一些實施例中，LNP包含約1.5 mol%至約2.0 mol%之聚合物偶聯脂質。在一些實施例中，LNP包含約1.5 mol%至約1.8 mol%之聚合物偶聯脂質。In some embodiments, the polymer-coupled lipid comprises about 0.5 mol% to about 5 mol% of the total lipid in the LNP. In some embodiments, the LNP comprises about 1.0 mol% to about 2.5 mol% of the polymer-coupled lipid. In some embodiments, the LNP comprises about 1.5 mol% to about 2.0 mol% of the polymer-coupled lipid. In some embodiments, the LNP comprises about 1.5 mol% to about 1.8 mol% of the polymer-coupled lipid.

在一些實施例中，總陽離子脂質與總聚合物偶聯脂質之莫耳比為約100:1至約20:1。在一些實施例中，總陽離子脂質與總聚合物偶聯脂質之莫耳比為約50:1至約20:1。在一些實施例中，總陽離子脂質與總聚合物偶聯脂質之莫耳比為約40:1至約20:1。在一些實施例中，總陽離子脂質與總聚合物偶聯脂質(例如，PEG偶聯脂質)之莫耳比為約35:1至約25:1。In some embodiments, the molar ratio of total cationic lipids to total polymer-coupled lipids is from about 100:1 to about 20:1. In some embodiments, the molar ratio of total cationic lipids to total polymer-coupled lipids is from about 50:1 to about 20:1. In some embodiments, the molar ratio of total cationic lipids to total polymer-coupled lipids is from about 40:1 to about 20:1. In some embodiments, the molar ratio of total cationic lipids to total polymer-coupled lipids (e.g., PEG-coupled lipids) is from about 35:1 to about 25:1.

(iii) 類固醇 如本文所述，在一些實施例中，核酸粒子進一步包含類固醇。在一些實施例中，類固醇係固醇。在一些實施例中，固醇係β-植物固醇、豆固醇、膽固醇、膽鈣化固醇、麥角鈣化固醇、卡泊三醇、肉毒毒素、羽扇豆醇、環阿屯醇、羊毛脂固醇或α-生育酚。在一些實施例中，固醇係β-植物固醇。在一些實施例中，固醇係豆固醇。在一些實施例中，固醇係膽固醇。在一些實施例中，固醇係膽鈣化固醇。在一些實施例中，固醇係麥角鈣化固醇。在一些實施例中，固醇係卡泊三醇。在一些實施例中，固醇係肉毒毒素。在一些實施例中，固醇係羽扇豆醇。在一些實施例中，固醇係環阿屯醇。在一些實施例中，固醇係羊毛脂固醇。在一些實施例中，固醇係α-生育酚。(iii) SteroidsAs described herein, in some embodiments, the nucleic acid particle further comprises a steroid. In some embodiments, the steroid is a sterol. In some embodiments, the sterol is β-phytosterol, stigmasterol, cholesterol, cholecalciferol, ergocalciferol, calcipotriol, botulinum toxin, lupeol, cycloartenol, lanolin sterol, or α-tocopherol. In some embodiments, the sterol is β-phytosterol. In some embodiments, the sterol is stigmasterol. In some embodiments, the sterol is cholesterol. In some embodiments, the sterol is cholecalciferol. In some embodiments, the sterol is ergocalciferol. In some embodiments, the sterol is calcipotriol. In some embodiments, the sterol is botulinum toxin. In some embodiments, the sterol is lupeol. In some embodiments, the sterol is cycloartenol. In some embodiments, the sterol is lanolin sterol. In some embodiments, the sterol is α-tocopherol.

在一些實施例中，醫藥組合物包含約30 mol%至約50 mol%之類固醇。在一些實施例中，醫藥組合物包含約35 mol%至約45 mol%之類固醇。在一些實施例中，醫藥組合物包含約38 mol%至約40 mol%之類固醇。在一些實施例中，醫藥組合物包含約38.5 mol%之類固醇。在一些實施例中，醫藥組合物包含約40 mol%之類固醇。In some embodiments, the pharmaceutical composition comprises about 30 mol% to about 50 mol% of the steroid. In some embodiments, the pharmaceutical composition comprises about 35 mol% to about 45 mol% of the steroid. In some embodiments, the pharmaceutical composition comprises about 38 mol% to about 40 mol% of the steroid. In some embodiments, the pharmaceutical composition comprises about 38.5 mol% of the steroid. In some embodiments, the pharmaceutical composition comprises about 40 mol% of the steroid.

在一些實施例中，醫藥組合物包含約30 mol%至約50 mol%之膽固醇。在一些實施例中，醫藥組合物包含約35 mol%至約45 mol%之膽固醇。在一些實施例中，醫藥組合物包含約38 mol%至約41 mol%之膽固醇。在一些實施例中，醫藥組合物包含約38.5 mol%之膽固醇。在一些實施例中，醫藥組合物包含約40.7 mol%之膽固醇。In some embodiments, the pharmaceutical composition comprises about 30 mol% to about 50 mol% cholesterol. In some embodiments, the pharmaceutical composition comprises about 35 mol% to about 45 mol% cholesterol. In some embodiments, the pharmaceutical composition comprises about 38 mol% to about 41 mol% cholesterol. In some embodiments, the pharmaceutical composition comprises about 38.5 mol% cholesterol. In some embodiments, the pharmaceutical composition comprises about 40.7 mol% cholesterol.

(iv) 陰離子兩親物 如本文所述，在一些實施例中，醫藥組合物包含陰離子兩親物。「兩親物」應理解為具有親水性部分及親脂性部分之分子。「親脂性」部分係可溶於非極性溶劑(例如己烷、四氫呋喃(THF)及/或氯仿)中之部分。「親水性」部分係可溶於水溶液(例如水)中之部分。(iv) Anionic AmphiphilesAs described herein, in some embodiments, the pharmaceutical compositions comprise an anionic amphiphile. "Amphiphile" is understood to mean a molecule having a hydrophilic portion and a lipophilic portion. The "lipophilic" portion is one that is soluble in non-polar solvents (e.g., hexane, tetrahydrofuran (THF), and/or chloroform). The "hydrophilic" portion is one that is soluble in aqueous solutions (e.g., water).

在一些實施例中，陰離子兩親物在特定pH (例如生理pH)下在兩親物之親水性部分中包含負電荷。在一些實施例中，陰離子兩親物為二羧酸與二醯基甘油之半酯。在一些實施例中，二醯基甘油部分之醯基部分之烴基部分為具有6至40個、較佳地8至18個碳原子之烷基(如上文所定義)或具有6至40個、較佳14至18個碳原子之烯基(如上文所定義)。在一些實施例中，醯基部分存在於甘油部分之1位及2位上。在一些實施例中，二醯基甘油部分之醯基部分係相同的。在一些實施例中，二醯基甘油部分之醯基部分係不同的。在一些實施例中，醯基部分為飽和脂肪酸部分，較佳選自由以下組成之群：硬脂醯基、棕櫚醯基、肉荳蔻醯基、月桂醯基、癸醯基及辛醯基部分。In some embodiments, the anionic amphiphile comprises a negative charge in the hydrophilic portion of the amphiphile at a specific pH (e.g., physiological pH). In some embodiments, the anionic amphiphile is a half ester of a dicarboxylic acid and a diacylglycerol. In some embodiments, the alkyl moiety of the acyl moiety of the diacylglycerol moiety is an alkyl group (as defined above) having 6 to 40, preferably 8 to 18, carbon atoms, or an alkenyl group (as defined above) having 6 to 40, preferably 14 to 18, carbon atoms. In some embodiments, the acyl moiety is present at the 1-position and the 2-position of the glycerol moiety. In some embodiments, the acyl moieties of the diacylglycerol moiety are identical. In some embodiments, the acyl moieties of the diacylglycerol moiety are different. In some embodiments, the acyl moiety is a saturated fatty acid moiety, preferably selected from the group consisting of stearyl, palmitoyl, myristyl, lauryl, caprylyl, and octanoyl moieties.

在一些實施例中，醯基部分為不飽和脂肪酸部分，較佳選自由油醯基、亞麻基及亞麻醯基部分組成之群。在一些實施例中，二羧酸部分具有2至20個碳原子、2至10個碳原子或2至8個碳原子。二羧酸部分之實例包括草酸、丙二酸、琥珀酸、戊二酸、己二酸、庚二酸及辛二酸。In some embodiments, the acyl moiety is an unsaturated fatty acid moiety, preferably selected from the group consisting of oleyl, linalool, and linalool moieties. In some embodiments, the dicarboxylic acid moiety has 2 to 20 carbon atoms, 2 to 10 carbon atoms, or 2 to 8 carbon atoms. Examples of dicarboxylic acid moieties include oxalic acid, malonic acid, succinic acid, glutaric acid, adipic acid, pimelic acid, and suberic acid.

在一些實施例中，帶負電荷之兩親物為琥珀酸與二醯基甘油之半酯(亦即，二羧酸部分為琥珀酸部分)，在本文中亦稱為「二醯基甘油半琥珀酸酯」。In some embodiments, the negatively charged amphiphile is a half-ester of succinic acid and diacylglycerol (ie, the dicarboxylic acid moiety is a succinic acid moiety), also referred to herein as "diacylglycerol hemisuccinate."

在一些實施例中，陰離子兩親物選自1,2-二月桂醯基甘油基半琥珀酸酯(DLGS)、1,2-二肉荳蔻醯基甘油基半琥珀酸酯(DMGS)、1,2-二棕櫚醯基甘油基半琥珀酸酯(DPGS)、1-棕櫚醯基-2-硬脂醯基甘油基半琥珀酸酯(PSGS)、二硬脂醯基半琥珀酸甘油酯(DSGS)、1,2-二油醯基半琥珀酸甘油酯(DOGS)、1-硬脂醯基2-肉荳蔻醯基-半琥珀酸甘油酯(SMGS)、1-棕櫚醯基-2-油醯基半琥珀酸甘油酯(POGS)及其上文中任一者之類似物，其中二羧酸部分為草酸、丙二酸、琥珀酸、戊二酸、己二酸、庚二酸或辛二酸。在一些實施例中，陰離子兩親物為二肉荳蔻醯基甘油基半琥珀酸酯、二棕櫚醯基甘油基半琥珀酸酯或二硬脂醯基甘油基半琥珀酸酯。。在一些實施例中，陰離子兩親物為熊果酸、齊墩果酸。In some embodiments, the anionic amphiphile is selected from 1,2-dilaurylglyceryl hemisuccinate (DLGS), 1,2-dimyristylglyceryl hemisuccinate (DMGS), 1,2-dipalmitoylglyceryl hemisuccinate (DPGS), 1-palmitoyl-2-stearoylglyceryl hemisuccinate (PSGS), distearyl hemisuccinate (PSGS), 1,2-dilaurylglyceryl hemisuccinate (DLGS), 1,2-dimyristylglyceryl hemisuccinate (DMGS), 1,2-dipalmitoylglyceryl hemisuccinate (DPGS), 1,2-dipalmitoylglyceryl hemisuccinate (PSGS), 1,2-dicarboxylic acid glyceryl hemisuccinate (DP ... (DSGS), 1,2-dioleyl hemisuccinate (DOGS), 1-stearyl-2-myristyl-hemisuccinate (SMGS), 1-palmitoyl-2-oleyl hemisuccinate (POGS), and analogs thereof, wherein the dicarboxylic acid moiety is oxalic acid, malonic acid, succinic acid, glutaric acid, adipic acid, pimelic acid, or suberic acid. In some embodiments, the anionic amphiphile is dimyristyl glyceryl hemisuccinate, dimalmitoyl glyceryl hemisuccinate, or distearyl glyceryl hemisuccinate. In some embodiments, the anionic amphiphile is ursolic acid or azinoic acid.

在一些實施例中，陰離子兩親物為二羧酸與膽固醇之半酯。在一些實施例中，陰離子兩親物選自下表：半琥珀酸膽固醇酯半己二酸膽固醇酯半辛二酸膽固醇酯半(癸二酸)膽固醇酯半(十二烷二酸)膽固醇酯半(十四烷二酸)膽固醇酯半(十六烷二酸)膽固醇酯半(十八烷二酸)膽固醇酯In some embodiments, the anionic amphiphile is a half ester of a dicarboxylic acid and cholesterol. In some embodiments, the anionic amphiphile is selected from the following table: Cholesterol hemisuccinate Cholesterol hemiadipate Cholesterol hemisuberate Cholesterol hemi(sebacic acid) ester Cholesterol hemi(dodecanedioate) Cholesterol hemi(tetradecanedioate) Cholesterol hemi(hexadecandioic acid) ester Cholesterol hemi(octadecane dioate)

在一些實施例中，本文所述之脂質奈米粒子包含約0.5 mol%至約10 mol%之陰離子兩親物。In some embodiments, the lipid nanoparticles described herein comprise about 0.5 mol % to about 10 mol % of anionic amphiphile.

在一些實施例中，本文所述之醫藥組合物包含： 約30 mol%至約60 mol%之化合物(例如式I、II-1、II-2、II-3或II-4之陽離子可離子化脂質)； 約20 mol%至約60 mol%之類固醇； 約1 mol%至約2 mol%之聚合物偶聯脂質；及 約5 mol%至約20 mol%之中性脂質。In some embodiments, the pharmaceutical compositions described herein comprise:about 30 mol% to about 60 mol% of a compound (e.g., a cationically ionizable lipid of Formula I, II-1, II-2, II-3, or II-4);about 20 mol% to about 60 mol% of a steroid;about 1 mol% to about 2 mol% of a polymer-conjugated lipid; andabout 5 mol% to about 20 mol% of a neutral lipid.

在一些實施例中，本文所述之醫藥組合物包含： 約30 mol%至約60 mol%之化合物(例如式I、II-1、II-2、II-3或II-4之陽離子可離子化脂質)； 約20 mol%至約60 mol%之類固醇； 約0.5至約10 mol%之陰離子兩親物；及 約5 mol%至約20 mol%之中性脂質。In some embodiments, the pharmaceutical compositions described herein comprise:about 30 mol% to about 60 mol% of a compound (e.g., a cationically ionizable lipid of Formula I, II-1, II-2, II-3, or II-4);about 20 mol% to about 60 mol% of a steroid;about 0.5 to about 10 mol% of an anionic amphiphile; andabout 5 mol% to about 20 mol% of a neutral lipid.

在一些實施例中，本文所述之醫藥組合物包含： 約30 mol%至約60 mol%之化合物(例如式I、II-1、II-2、II-3或II-4之陽離子可離子化脂質)； 約20 mol%至約60 mol%之類固醇； 約0.5至約10 mol%之陰離子兩親物；及 約5 mol%至約20 mol%之中性脂質，且進一步其中該醫藥組合物不包括聚合物偶聯脂質。In some embodiments, the pharmaceutical compositions described herein comprise:about 30 mol% to about 60 mol% of a compound (e.g., a cationically ionizable lipid of Formula I, II-1, II-2, II-3, or II-4);about 20 mol% to about 60 mol% of a steroid;about 0.5 to about 10 mol% of an anionic amphiphile; andabout 5 mol% to about 20 mol% of a neutral lipid, further wherein the pharmaceutical composition does not include a polymer-conjugated lipid.

術語「平均直徑(average diameter)」或「平均直徑(mean diameter)」係指如藉由動態雷射光散射(DLS)量測之粒子之平均流體力學直徑，使用所謂的累積量演算法進行數據分析，其提供所謂的具有長度維度之Z平均值及無維度之多分散性指數(PDI)作為結果(Koppel, D., J. Chem. Phys. 57, 1972，第4814-4820頁，ISO 13321)。在此處，粒子之「平均直徑(average diameter)」、「平均直徑(mean diameter)」、「直徑」或「大小」與Z平均值之此值以同義使用。The term "average diameter" or "mean diameter" refers to the mean hydrodynamic diameter of particles, as measured by dynamic laser light scattering (DLS). Data analysis using the so-called cumulative volume algorithm provides the so-called Z-average value, which has a length dimension, and the dimensionless polydispersity index (PDI) as a result (Koppel, D., J. Chem. Phys. 57, 1972, pp. 4814-4820, ISO 13321). Herein, the terms "average diameter," "mean diameter," "diameter," or "size" of particles are used synonymously with this value of the Z-average value.

「多分散性指數」較佳地基於動態光散射量測藉由如「平均直徑」之定義中所提及之所謂的累積量分析來計算。在某些先決條件下，其可作為核糖核酸奈米粒子(例如，核糖核酸奈米粒子)整體之大小分佈之量度。The "polydispersity index" is preferably calculated based on dynamic light scattering measurements by so-called cumulative mass analysis as mentioned in the definition of "mean diameter." Under certain prerequisites, it can be used as a measure of the size distribution of the RNA nanoparticle population (e.g., RNA nanoparticles).

不同類型之核酸粒子先前已描述為適於遞送呈顆粒形式之核酸(例如，Kaczmarek, J. C.等人，2017, Genome Medicine 9, 60)。對於非病毒核酸遞送媒劑，核酸之奈米粒子囊封在物理上保護核酸免於降解，且端視特定化學可幫助細胞攝取及胞內體逃逸。Various types of nucleic acid particles have been previously described as suitable for delivering nucleic acids in particulate form (e.g., Kaczmarek, J. C. et al., 2017, Genome Medicine 9, 60). For non-viral nucleic acid delivery vehicles, nanoparticle encapsulation of nucleic acids physically protects the nucleic acids from degradation and, depending on the specific chemistry, can facilitate cellular uptake and endosomal escape.

本文所述之一些實施例係關於涉及一種以上、例如2種、3種、4種、5種、6種或甚至更多種核酸種類之組合物、方法及用途。核酸種類可為RNA及/或DNA。舉例而言，本文所述之粒子可含有一種RNA (例如，一種mRNA)及一種DNA。Some embodiments described herein relate to compositions, methods, and uses involving more than one, for example, two, three, four, five, six, or even more, nucleic acid species. The nucleic acid species can be RNA and/or DNA. For example, a particle described herein can contain one RNA (e.g., one mRNA) and one DNA.

在核酸粒子組合物中，每一核酸種類單獨調配為個別核酸粒子調配物係可能的。在該情形下，每一個別核酸粒子調配物將包含一種核酸種類。個別核酸粒子調配物可作為單獨實體存在於例如單獨容器中。該等調配物可藉由單獨提供每一核酸種類(通常各自呈含核酸之溶液形式)以及粒子形成劑來獲得，由此允許形成粒子。各別粒子將僅含在形成粒子(個別粒子調配物)時提供之特定核酸種類。In nucleic acid particle compositions, it is possible to separately formulate each nucleic acid species as a separate nucleic acid particle formulation. In this case, each separate nucleic acid particle formulation would contain a single nucleic acid species. The separate nucleic acid particle formulations can be present as separate entities, for example, in separate containers. Such formulations can be obtained by separately providing each nucleic acid species (typically each in the form of a nucleic acid-containing solution) and a particle-forming agent, thereby allowing particle formation. Each individual particle will contain only the specific nucleic acid species provided during particle formation (the individual particle formulation).

(v) 製造脂質奈米粒子之方法 脂質及包含核酸之脂質奈米粒子以及其製備方法為此項技術中已知，包括例如如美國專利第8,569,256號、第5,965,542號及美國專利公開案第2016/0199485號、第2016/0009637號、第2015/0273068號、第2015/0265708號、第2015/0203446號、第2015/0005363號、第2014/0308304號、第2014/0200257號、第2013/086373號、第2013/0338210號、第2013/0323269號、第2013/0245107號、第2013/0195920號、第2013/0123338號、第2013/0022649號、第2013/0017223號、第2012/0295832號、第2012/0183581號、第2012/0172411號、第2012/0027803號、第2012/0058188號、第2011/0311583號、第2011/0311582號、第2011/0262527號、第2011/0216622號、第2011/0117125號、第2011/0091525號、第2011/0076335號、第2011/0060032號、第2010/0130588號、第2007/0042031號、第2006/0240093號、第2006/0083780號、第2006/0008910號、第2005/0175682號、第2005/017054號、第2005/0118253號、第2005/0064595號、第2004/0142025號、第2007/0042031號、第1999/009076號及PCT公開案第WO 99/39741號、第WO 2018/081480號、第WO 2017/004143號、第WO 2017/075531號、第WO 2015/199952號、第WO 2014/008334號、第WO 2013/086373號、第WO 2013/086322號、第WO 2013/016058號、第WO 2013/086373號、第W02011/141705號及第WO 2001/07548號中所述，該等專利之全部揭示內容之全文出於本文所述之目的皆以引用方式併入本文中。(v) Methods for Making Lipid NanoparticlesLipids and nucleic acid-containing lipid nanoparticles and methods for preparing the same are known in the art, including, for example, U.S. Patent Nos. 8,569,256 and 5,965,542, and U.S. Patent Publication Nos. 2016/0199485, 2016/0009637, 2015/0273068, 2015/0265708, 2015/0203446, 2015/0005363, and 2014/030830. 4, 2014/0200257, 2013/086373, 2013/0338210, 2013/0323269, 2013/0245107, 2013/0195920, 2013/0123338, 2013/0022649, 2013/0017223, 2012/0295832, 2012/0183581, 2012/ No. 0172411, No. 2012/0027803, No. 2012/0058188, No. 2011/0311583, No. 2011/0311582, No. 2011/0262527, No. 2011/0216622, No. 2011/0117125, No. 2011/0091525, No. 2011/0076335, No. 2011/0060032, No. 2010/0130588 , 2007/0042031, 2006/0240093, 2006/0083780, 2006/0008910, 2005/0175682, 2005/017054, 2005/0118253, 2005/0064595, 2004/0142025, 2007/0042031, 1999/009076, and PCT Publication No. WO 99/39741, WO 2018/081480, WO 2017/004143, WO 2017/075531, WO 2015/199952, WO 2014/008334, WO 2013/086373, WO 2013/086322, WO 2013/016058, WO 2013/086373, WO 2011/141705, and WO 2001/07548, the entire disclosures of which are incorporated herein by reference for the purposes herein.

舉例而言，在一些實施例中，可用於遞送核酸之脂質以預定重量或莫耳比/百分比(例如，本文所述之重量或莫耳比/百分比)溶解於乙醇中。在一些實施例中，脂質奈米粒子(LNP)係以大約6:1至30:1之總脂質對RNA或DNA莫耳比製備。在一些實施例中，該RNA或DNA可在乙酸鹽緩衝液中稀釋至0.2 mg/mL。For example, in some embodiments, lipids useful for nucleic acid delivery can be dissolved in ethanol at a predetermined weight or molar ratio/percentage (e.g., the weights or molar ratios/percentages described herein). In some embodiments, lipid nanoparticles (LNPs) are prepared at a total lipid to RNA or DNA molar ratio of approximately 6:1 to 30:1. In some embodiments, the RNA or DNA can be diluted to 0.2 mg/mL in acetate buffer.

在一些實施例中，使用乙醇注射技術，可如下形成包含核酸(例如，RNA或DNA)之膠體脂質分散液：將包含脂質(諸如本文所述之環狀可離子化脂質、中性脂質、類固醇以及視情況選用之聚合物偶聯脂質或陰離子兩親物)之乙醇溶液注射至包含核酸(例如，RNA或DNA)之水溶液中。In some embodiments, colloidal lipid dispersions containing nucleic acids (e.g., RNA or DNA) can be formed using an ethanol injection technique by injecting an ethanol solution containing lipids (such as cyclic ionizable lipids, neutral lipids, steroids, and optionally polymer-conjugated lipids or anionic amphiphiles as described herein) into an aqueous solution containing nucleic acids (e.g., RNA or DNA).

在一些實施例中，可在室溫下藉由例如使用活塞泵將每一溶液(例如，包含本文所述之環狀可離子化脂質化合物、中性脂質、類固醇以及視情況選用之聚合物偶聯脂質或陰離子兩親物或任何其他添加劑之脂質溶液)以受控流量泵送至混合單元中來混合脂質及核酸溶液。在一些實施例中，脂質溶液及核酸溶液進入混合單元中之流量維持在1:3之比率。混合後，當用水性RNA稀釋乙醇脂質溶液時，形成核酸-脂質粒子。脂質溶解度減小，同時帶有正電荷之陽離子脂質與帶負電核酸相互作用。In some embodiments, the lipid and nucleic acid solutions can be mixed at room temperature by pumping each solution (e.g., a lipid solution comprising a cyclic ionizable lipid compound described herein, a neutral lipid, a steroid, and optionally a polymer-conjugated lipid or anionic amphiphile, or any other additive) at a controlled flow rate into a mixing unit, for example, using a piston pump. In some embodiments, the flow rate of the lipid solution and the nucleic acid solution into the mixing unit is maintained at a ratio of 1:3. After mixing, when the ethanolic lipid solution is diluted with aqueous RNA, nucleic acid-lipid particles are formed. The lipid solubility decreases, and the positively charged cationic lipids interact with the negatively charged nucleic acids.

在一些實施例中，包含囊封核酸(例如，RNA)之脂質奈米粒子之溶液可藉由濃度調節、緩衝液交換、調配及/或過濾中之一或多者來處理。In some embodiments, a solution comprising lipid nanoparticles encapsulating nucleic acids (e.g., RNA) can be processed by one or more of concentration adjustment, buffer exchange, blending, and/or filtration.

在一些實施例中，本文所述之組合物或複合物進一步包含醫藥學上可接受之表面活性劑。在一些實施例中，醫藥學上可接受之表面活性劑選自聚山梨醇酯(例如，聚山梨醇酯20 (Tween20)、聚山梨醇酯40 (Tween40)、聚山梨醇酯60 (Tween60)及聚山梨醇酯80 (Tween80))、泊洛沙姆(poloxamer)以及包含選自聚烷二醇(例如，聚乙二醇)、聚(2-噁唑啉)、聚(2-甲基-2-噁唑啉)、聚肌胺酸、聚乙烯基吡咯啶酮及聚[N-(2-羥基丙基)甲基丙烯醯胺之部分之兩親性基團，其中該部分結合至一或多個C₁₂-C₂₀脂族基團。In some embodiments, the compositions or complexes described herein further comprise a pharmaceutically acceptable surfactant. In some embodiments, the pharmaceutically acceptable surfactant is selected from polysorbates (e.g., polysorbate 20 (Tween 20), polysorbate 40 (Tween 40), polysorbate 60 (Tween 60), and polysorbate 80 (Tween 80)), poloxamers, and amphiphilic groups comprising a moiety selected from polyalkylene glycols (e.g., polyethylene glycol), poly(2-oxazoline), poly(2-methyl-2-oxazoline), polysarcosine, polyvinylpyrrolidone, and poly[N-(2-hydroxypropyl)methacrylamide], wherein the moiety is bound to one or more C₁₂ -C₂₀ aliphatic groups.

RNA在一些實施例中，本文所述之粒子包含一或多種寡糖組合物及核酸。在一些實施例中，核酸係RNA。RNA In some embodiments, the particles described herein comprise one or more oligosaccharide compositions and a nucleic acid. In some embodiments, the nucleic acid is RNA.

在一些實施例中，適於本文所述技術之RNA係單股RNA。在一些實施例中，如本文所揭示之RNA係線性RNA。在一些實施例中，單股RNA係非編碼RNA之原因在於，其核苷酸序列不包括開放閱讀框(或其補體)。在一些實施例中，單股RNA具有編碼本揭示案之多肽或複數個多肽(例如，抗原決定基)之核苷酸序列(或係編碼該多肽或複數個多肽之序列之補體)。In some embodiments, RNA suitable for use with the techniques described herein is single-stranded RNA. In some embodiments, RNA as disclosed herein is linear RNA. In some embodiments, a single-stranded RNA is non-coding RNA because its nucleotide sequence does not include an open reading frame (or a complement thereof). In some embodiments, a single-stranded RNA has a nucleotide sequence that encodes a polypeptide or polypeptides (e.g., an antigenic determinant) of the present disclosure (or is a complement of a sequence that encodes the polypeptide or polypeptides).

在一些實施例中，RNA係或包含siRNA、miRNA或其他非編碼RNA。In some embodiments, the RNA is or comprises siRNA, miRNA, or other non-coding RNA.

在許多實施例中，相關RNA包括至少一個開放閱讀框(ORF) (例如，係mRNA)；在一些實施例中，相關RNA包括單一ORF；在一些實施例中，相關RNA包括多於一個ORF。In many embodiments, the related RNA comprises at least one open reading frame (ORF) (e.g., is an mRNA); in some embodiments, the related RNA comprises a single ORF; in some embodiments, the related RNA comprises more than one ORF.

在一些實施例中，RNA包含例如編碼所關注多肽或編碼所關注之複數個多肽之ORF。在一些實施例中，根據本文所提供之技術產生之RNA包含複數個ORF (例如，編碼複數個多肽)。在一些實施例中，根據本文技術產生之RNA包含編碼複數個多肽之單一ORF。在一些此類實施例中，多肽係或包含抗原或其抗原決定基(例如，相關抗原)。In some embodiments, the RNA comprises an ORF,e.g., encoding a polypeptide of interest or encoding multiple polypeptides of interest. In some embodiments, the RNA produced according to the techniques provided herein comprises multiple ORFs (e.g., encoding multiple polypeptides). In some embodiments, the RNA produced according to the techniques provided herein comprises a single ORF encoding multiple polypeptides. In some such embodiments, the polypeptide comprises an antigen or an antigenic determinant thereof (e.g., a related antigen).

在一些實施例中，根據本揭示案使用之ORF編碼包括例如在哺乳動物細胞中起作用之信號序列(例如固有信號序列或異源信號序列)之多肽。在一些實施例中，信號序列指導經編碼多肽之分泌，在一些實施例中，信號序列指導經編碼多肽輸送至所界定細胞隔室、較佳地細胞表面、內質網(ER)或胞內體-溶酶體隔室中。In some embodiments, the ORF used in accordance with the present disclosure encodes a polypeptide that includes a signal sequence (e.g. , a native signal sequence or a heterologous signal sequence) that functions in mammalian cells. In some embodiments, the signal sequence directs secretion of the encoded polypeptide, and in some embodiments, the signal sequence directs trafficking of the encoded polypeptide to a defined cellular compartment, preferably the cell surface, the endoplasmic reticulum (ER), or the endosomal-lysosomal compartment.

在一些實施例中，ORF編碼包括多聚元件(例如，固有或異源多聚元件)之多肽。在一些實施例中，編碼表面多肽(例如，包括指導表面定位之信號序列之多肽)之ORF包括多聚元件。In some embodiments, an ORF encodes a polypeptide comprising a multimerization element (e.g., an intrinsic or heterologous multimerization element). In some embodiments, an ORF encoding a surface polypeptide (e.g., a polypeptide comprising a signal sequence directing surface localization) comprises a multimerization element.

在一些實施例中，ORF編碼包括跨膜元件或結構域之多肽。In some embodiments, the ORF encodes a polypeptide that includes a transmembrane element or domain.

在一些實施例中，ORF經密碼子最佳化以在特定宿主(例如哺乳動物宿主，例如人類)之細胞中表現。In some embodiments, the ORF is codon-optimized for expression in the cells of a specific host (e.g., a mammalian host,such as a human).

在一些實施例中，RNA包括未經修飾之尿苷殘基；僅包括未經修飾之尿苷殘基之RNA可稱為「uRNA」。在一些實施例中，RNA包括一或多個經修飾之尿苷殘基；在一些實施例中，該RNA (例如，包括完全經修飾之尿苷殘基之RNA)稱為「modRNA」。在一些實施例中，RNA可為自擴增RNA (saRNA)。在一些實施例中，RNA可為反式擴增RNA (taRNA) (參見例如WO2017/162461)。In some embodiments, the RNA includes unmodified uridine residues; RNA that includes only unmodified uridine residues may be referred to as "uRNA." In some embodiments, the RNA includes one or more modified uridine residues; in some embodiments, the RNA (e.g., RNA that includes all modified uridine residues) is referred to as "modRNA." In some embodiments, the RNA may be self-amplifying RNA (saRNA). In some embodiments, the RNA may be trans-amplifying RNA (taRNA) (see , e.g., WO2017/162461).

在一些實施例中，相關RNA包括多肽編碼部分或複數個多肽編碼部分。在一些具體實施例中，該一或多個部分可編碼一或多種多肽，其係或包含生物活性多肽或其部分(例如，酶或細胞介素或治療蛋白，例如替代蛋白或其抗體或部分)。在一些具體實施例中，該一或多個部分可編碼一或多種多肽，其係或包含抗原(或其抗原決定基)、細胞介素、酶等。在一些實施例中，一或多種經編碼多肽可為或包括與腫瘤相關之一或多種新抗原或新抗原決定基。在一些實施例中，一或多種經編碼多肽可為或包括傳染原(例如，細菌、真菌、病毒等)之一或多種抗原(或其抗原決定基)。在某些實施例中，經編碼多肽可為野生型多肽之變異體。In some embodiments, the RNA of interest comprises a polypeptide-encoding portion or portions thereof. In some embodiments, the one or more portions may encode one or more polypeptides, which are or comprise biologically active polypeptides or portions thereof (e.g., enzymes or interleukins or therapeutic proteins, such as replacement proteins or antibodies or portions thereof). In some embodiments, the one or more portions may encode one or more polypeptides, which are or comprise antigens (or antigenic determinants thereof), interleukins, enzymes, etc. In some embodiments, the one or more encoded polypeptides may be or comprise one or more neoantigens or neoantigens associated with tumors. In some embodiments, the one or more encoded polypeptides may be or comprise one or more antigens (or antigenic determinants thereof) of infectious agents (e.g. , bacteria, fungi, viruses, etc.). In certain embodiments, the encoded polypeptides may be variants of wild-type polypeptides.

在一些實施例中，單股RNA (例如，mRNA)可包含分泌信號編碼區(例如，允許一或多種經編碼靶實體在經細胞轉譯後分泌之分泌信號編碼區)。在一些實施例中，該分泌信號編碼區可為或包含非人類分泌信號。在一些實施例中，該分泌信號編碼區可為或包含人類分泌信號。In some embodiments, a single-stranded RNA (e.g., mRNA) may comprise a secretory signal coding region (e.g., a secretory signal coding region that allows one or more encoded target entities to be secreted after translation by a cell). In some embodiments, the secretory signal coding region may be or comprise a non-human secretory signal. In some embodiments, the secretory signal coding region may be or comprise a human secretory signal.

在一些實施例中，單股RNA (例如，mRNA)可包含至少一種非編碼元件(例如，以增強RNA穩定性及/或轉譯效率)。非編碼元件之實例包括(但不限於) 3’非轉譯區(UTR)、5’ UTR、帽結構(例如，在一些實施例中，酶添加之帽；在一些實施例中，共轉錄帽)、聚腺嘌呤(聚A)尾(例如，在一些實施例中，其可為或包含100或更多個A殘基，及/或在一些實施例中，其可包括一或多個「中斷」 [亦即，非A]序列元件)及其任何組合。該等非編碼元件之示範性實施例可參見例如WO2011015347、WO2017053297、US 10519189、US 10494399、WO2007024708、WO2007036366、WO2017060314、WO2016005324、WO2005038030、WO2017036889、WO2017162266及WO2017162461，該等專利中每一者之全文皆以引用方式併入本文中。In some embodiments, a single-stranded RNA (e.g. , mRNA) may comprise at least one non-coding element (e.g. , to enhance RNA stability and/or translation efficiency). Examples of non-coding elements include, but are not limited to, a 3' untranslated region (UTR), a 5' UTR, a cap structure (e.g., in some embodiments, an enzymatically added cap; in some embodiments, a co-transcriptional cap), a polyadenine (poly-A) tail (e.g., in some embodiments, it may be or comprise 100 or more A residues, and/or in some embodiments, it may include one or more "interrupted" [i.e., non-A] sequence elements), and any combination thereof. Exemplary embodiments of these non-coding elements can be found in, for example, WO2011015347, WO2017053297, US 10519189, US 10494399, WO2007024708, WO2007036366, WO2017060314, WO2016005324, WO2005038030, WO2017036889, WO2017162266, and WO2017162461, the entire contents of each of which are incorporated herein by reference.

格式 已開發出可用於RNA醫藥組合物(例如，免疫原性組合物或疫苗)之至少四種格式，即含未經修飾尿苷之mRNA (uRNA)、核苷修飾之mRNA (modRNA)、自擴增mRNA (saRNA)及反式擴增RNA。FormatsAt least four formats have been developed for use in RNA pharmaceutical compositions (e.g., immunogenic compositions or vaccines): unmodified uridine-containing mRNA (uRNA), nucleoside-modified mRNA (modRNA), self-amplifying mRNA (saRNA), and trans-amplifying RNA.

未經修飾之尿苷平臺之特徵可包括例如固有佐劑效應、良好的耐受性及安全性以及強抗體及T細胞反應中之一或多者。Characteristics of the unmodified uridine platform may include, for example, one or more of an intrinsic adjuvant effect, good tolerability and safety, and strong antibody and T cell responses.

經修飾尿苷(例如，假尿苷)平臺之特徵可包括降低的佐劑效應、鈍化的免疫先天免疫感測活化能力及因此加強的抗原表現、良好的耐受性及安全性以及強抗體及CD4-T細胞反應。如本文所述，本揭示案提供該等強抗體及CD4 T細胞反應尤其可用於疫苗接種之見解。Characteristics of the modified uridine (e.g., pseudouridine) platform may include reduced adjuvant effects, blunted activation of innate immune sensing and thus enhanced antigen presentation, good tolerability and safety, and strong antibody and CD4-T cell responses. As described herein, the present disclosure provides insights that these strong antibody and CD4 T cell responses are particularly useful for vaccination.

自擴增平臺之特徵可包括例如多肽(例如，蛋白質)表現之持續時間長、良好的耐受性及安全性、使用極低疫苗劑量產生功效之較高可能性。Characteristics of a self-expanding platform may include, for example, prolonged expression of the polypeptide (e.g., protein), good tolerability and safety, and a higher likelihood of efficacy using very low vaccine doses.

在一些實施例中，自擴增平臺(例如，saRNA)包含編碼複製酶(例如，病毒複製酶)及所關注基因之核酸分子，其中核酸分子能夠由該複製酶順式複製(順式複製系統)。在一些實施例中，反式擴增平臺(例如，taRNA)包含兩種核酸分子，其中一種核酸分子編碼複製酶(例如，病毒複製酶)且另一核酸分子能夠由該複製酶反式複製(例如，複製子) (反式複製系統)。在一些實施例中，自擴增/反式擴增平臺(例如，RNA)包含複數種核酸分子，其中該等核酸編碼複數種複製酶及/或複製子。In some embodiments, a self-amplifying platform (e.g., saRNA ) comprises a nucleic acid molecule encoding a replicase (e.g., a viral replicase) and a gene of interest, wherein the nucleic acid molecule is capable of being replicatedin tandem by the replicase (a tandem replication system). In some embodiments, a trans-amplifying platform (e.g., taRNA ) comprises two nucleic acid molecules, one of which encodes a replicase (e.g., a viral replicase) and the other of which is capable of being replicatedin trans by the replicase (e.g., a replicon) (atrans- replication system). In some embodiments, a self-amplifying/trans-amplifying platform (e.g., RNA) comprises a plurality of nucleic acid molecules, wherein the nucleic acids encode a plurality of replicase and/or replicons.

在一些實施例中，反式複製系統包括在單一宿主細胞中存在兩種核酸分子。In some embodiments, thetrans- replication system comprises the presence of two nucleic acid molecules in a single host cell.

在一些此類實施例中，編碼複製酶(例如，病毒複製酶)之核酸無法在靶細胞及/或靶生物體中自複製。在一些此類實施例中，編碼複製酶(例如，病毒複製酶)之核酸缺少對基於(+)股模板合成(-)股及/或基於(-)股模板合成(+)股至關重要的至少一種保守序列元件。In some such embodiments, the nucleic acid encoding the replicase (e.g., a viral replicase) is unable to self-replicate in the target cell and/or target organism. In some such embodiments, the nucleic acid encoding the replicase (e.g., a viral replicase) lacks at least one conserved sequence element that is critical for the synthesis of (-) strands from a (+) strand template and/or the synthesis of (+) strands from a (-) strand template.

在一些實施例中，自擴增RNA包含5’帽；在一些反式複製系統中，編碼複製酶之至少RNA係加帽的。不希望受限於任一理論，已發現5’帽可能對所關注基因之反式高水準表現至關重要。In some embodiments, the self-amplifying RNA comprises a 5'cap; in sometrans- replication systems, at least the RNA encoding the replicase is capped. Without wishing to be bound by any theory, it has been discovered that the 5' cap may be crucial for high-level expression of the gene of interestin trans .

在一些實施例中，自擴增/反式擴增平臺不需要病毒粒子之繁殖(例如，與不期望病毒粒子形成無關)。在一些實施例中，自擴增/反式擴增平臺無法形成病毒粒子。In some embodiments, the self-amplification/trans-amplification platform does not require the propagation of viral particles (e.g., is not associated with undesirable viral particle formation). In some embodiments, the self-amplification/trans-amplification platform is unable to form viral particles.

在一些實施例中，RNA可包含內部核糖體進入位點(IRES)元件。在一些實施例中，RNA不包含IRES位點；具體而言，在一些實施例中，saRNA不包含IRES位點。在一些此類實施例中，所關注基因及/或複製酶之轉譯並非由IRES元件驅動。在一些實施例中，IRES元件經5’帽取代。在一些此類實施例中，經5’帽取代不會影響由RNA編碼之多肽序列。In some embodiments, the RNA may comprise an internal ribosome entry site (IRES) element. In some embodiments, the RNA does not comprise an IRES site; specifically, in some embodiments, the saRNA does not comprise an IRES site. In some such embodiments, translation of the gene of interest and/or replicase is not driven by an IRES element. In some embodiments, the IRES element is replaced with a 5' cap. In some such embodiments, replacement of the 5' cap does not affect the polypeptide sequence encoded by the RNA.

在一些實施例中，本文所述之複合物包含modRNA、saRNA、taRNA或uRNA。在一些實施例中，複合物包含modRNA。在一些實施例中，複合物包含saRNA。在一些實施例中，複合物包含taRNA。在一些實施例中，複合物包含uRNA。In some embodiments, the complexes described herein comprise modRNA, saRNA, taRNA, or uRNA. In some embodiments, the complexes comprise modRNA. In some embodiments, the complexes comprise saRNA. In some embodiments, the complexes comprise taRNA. In some embodiments, the complexes comprise uRNA.

使用方法本文所述之粒子可用於治療及預防個體之本文所述之疾病、病症及疾患。在一些實施例中，本揭示案提供治療疾病、病症或疾患之方法，其包括向患者投與包含本文所述粒子之組合物。在一些實施例中，本揭示案提供包含本文所述粒子之組合物之用途，其用於治療疾病、病症或疾患。在一些實施例中，疾病、病症或疾患係傳染病、癌症、自體免疫疾病或罕見疾病。Methods of Use The particles described herein can be used to treat and prevent the diseases, disorders, and conditions described herein in a subject. In some embodiments, the present disclosure provides methods of treating a disease, disorder, or condition comprising administering to a patient a composition comprising the particles described herein. In some embodiments, the present disclosure provides uses of compositions comprising the particles described herein for treating a disease, disorder, or condition. In some embodiments, the disease, disorder, or condition is an infectious disease, cancer, autoimmune disease, or rare disease.

在一些實施例中，傳染病係由病毒病原體引起或與其相關。在一些實施例中，病毒病原體屬於選自以下之科：痘病毒科(poxviridae)、彈狀病毒科(rhabdoviridae)、絲狀病毒科(filoviridae)、副黏液病毒科(paramyxoviridae)、嗜肝DNA病毒科(hepadnaviridae)、冠狀病毒科(coronaviridae)、杯狀病毒科(caliciviridae)、小核糖核酸病毒科(picornaviridae)、呼腸病毒科(reoviridae)、逆轉錄病毒科(retroviridae)及正黏液病毒科(orthomyxoviridae)。在一些實施例中，傳染病係由選自以下之病毒引起或與其相關：SARS-CoV-2、流感病毒、克里米亞剛果出血熱(Crimean-Congo Hemorhhagic Fever，CCHF)、伊波拉病毒(Ebola virus)、拉沙病毒(Lassa virus)、馬堡病毒(Marburg virus)、HIV、立百病毒(Nipah virus)及MERS-CoV。In some embodiments, the infectious disease is caused by or associated with a viral pathogen. In some embodiments, the viral pathogen belongs to a family selected from the group consisting of poxviridae, rhabdoviridae, filoviridae, paramyxoviridae, hepadnaviridae, coronaviridae, caliciviridae, picornaviridae, reoviridae, retroviridae, and orthomyxoviridae. In some embodiments, the infectious disease is caused by or is associated with a virus selected from the group consisting of SARS-CoV-2, influenza virus, Crimean-Congo Hemorrhagic Fever (CCHF), Ebola virus, Lassa virus, Marburg virus, HIV, Nipah virus, and MERS-CoV.

在一些實施例中，傳染病係由細菌病原體引起或與其相關。在一些實施例中，細菌病原體屬於選自以下之屬：以色列放線菌(Actinomyces israelii)、炭疽桿菌(bacillus antracis)、脆弱類桿菌(Bacteroides fragilis)、百日咳嗜血桿菌(Bordetella pertussis)、伯氏疏螺旋體(Borrelia burgdorferi)、伽氏疏螺旋體(Borrelia garinii)、包柔氏疏螺旋體(Borrelia afzelii)、迴歸熱螺旋體(Borrelia recurrentis)、流產布氏桿菌(Brucella abortus)、犬布氏桿菌(Brucella canis)、地中海熱布氏桿菌(Brucella melitensis)、豬布氏桿菌(Brucella suis)、空腸彎曲桿菌(Campolobacter jejuni)、肺炎披衣菌(Chlamydia pneumoniae)、沙眼披衣菌(Chlamydia trachomatis)、鸚鵡熱衣原體(Chlamydophila psittaci)、肉毒桿菌(Clostridium botulinum)、難養芽孢梭菌(Clostridium difficile)、產氣莢膜芽孢梭菌(Clostridium perfringens)、破傷風梭菌(Clostridium tetani)、白喉棒狀桿菌(Corynebacterium diphtheriae)、犬艾利希體(Ehrlichia canis)、查菲艾利希體(Ehrlichia chaffeensis)、糞腸球菌(Enterococcus faecalis)、屎腸球菌(Enterococcus faecium)、大腸桿菌(Escherichia coli)、土拉文氏桿菌(Francisella tularensis)、流感嗜血桿菌(Haemophilus influenzae)、幽門螺旋桿菌(Helicobacter pylori)、克雷伯氏肺炎桿菌(Klebsiella pneumoniae)、退伍軍人症嗜肺桿菌(Legionella pneumophila)、鉤端螺旋體(Leptospira)、李斯特單胞菌(Listeria monocytogenes)、麻瘋分枝桿菌(Mycobacterium leprae)、結核分枝桿菌(Mycobacterium tuberculosis)、肺炎黴漿菌(Mycoplasma pneumoniae)、淋病雙球菌(Neisseria gonorrhoeae)、腦膜炎雙球菌(Neisseria meningitidis)、銅綠假單胞菌(Pseudomonas aeruginosa)、星形諾卡菌(Nocardia asteroids)、立氏立克次體(Rickettsia ricektssii)、傷寒沙氏桿菌(Salmonella typhi)、鼠傷寒沙氏桿菌(Salmonella typhimurium)、宋內氏桿菌(Shigella sonnei)、赤痢桿菌(Shigella dysenteriae)、金黃色葡萄球菌(Staphylococcus aureus)、表皮葡萄球菌(Staphylococcus epidermidis)、腐生葡萄球菌(Staphylococcus saprophyticus)、無乳鏈球菌(Streptococcus agalactiae)、肺炎鏈球菌(Streptococcus pneumoniae)、釀膿鏈球菌(Streptococcus pyogenes)、草綠色鏈球菌(Streptococcus viridans)、梅毒螺旋體(Treponema pallidum)、霍亂弧菌(Vibrio cholerae)及鼠疫桿菌(Yersinia pestis)。In some embodiments, the infectious disease is caused by or associated with a bacterial pathogen. In some embodiments, the bacterial pathogen is of a genus selected from the group consisting of Actinomyces israelii, Bacillus antracis, Bacteroides fragilis, Bordetella pertussis, Borrelia burgdorferi, Borrelia garinii, Borrelia afzelii, Borrelia recurrentis, Brucella abortus, Brucella canis, Brucella melitensis, Brucella suis, Campolobacter jejuni, Chlamydia pneumoniae, pneumoniae), Chlamydia trachomatis, Chlamydophila psittaci, Clostridium botulinum, Clostridium difficile, Clostridium perfringens, Clostridium tetani, Corynebacterium diphtheriae, Ehrlichia canis, Ehrlichia chaffeensis, Enterococcus faecalis, Enterococcus faecium, Escherichia coli, Francisella tularensis tularensis), Haemophilus influenzae, Helicobacter pylori, Klebsiella pneumoniae, Legionella pneumophila, Leptospira, Listeria monocytogenes, Mycobacterium leprae, Mycobacterium tuberculosis, Mycoplasma pneumoniae, Neisseria gonorrhoeae, Neisseria meningitidis, Pseudomonas aeruginosa, Nocardia asteroids), Rickettsia ricektssii, Salmonella typhi, Salmonella typhimurium, Shigella sonnei, Shigella dysenteriae, Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcus saprophyticus, Streptococcus agalactiae, Streptococcus pneumoniae, Streptococcus pyogenes, Streptococcus viridans, Treponema pallidum, Vibrio cholerae cholerae) and Yersinia pestis.

在一些實施例中，傳染病係由寄生蟲引起或與其相關。在一些實施例中，寄生蟲屬於選自以下之科：瘧原蟲(Plasmodium)、利什曼原蟲屬(Leishmania)、隱孢子蟲屬(Cryptosporidium)、阿米巴屬(Entamoeba)、錐蟲屬(Trypanosomas)、血吸蟲(Schistosomes)、蛔蟲屬(Ascaris)、胞蟲屬(Echinococcus)及帶絛蟲屬(Taeniidae)。In some embodiments, the infectious disease is caused by or associated with a parasite. In some embodiments, the parasite belongs to a family selected from the group consisting of Plasmodium, Leishmania, Cryptosporidium, Entamoeba, Trypanosomas, Schistosomes, Ascaris, Echinococcus, and Taeniidae.

在一些實施例中，疾病、病症或疾患係癌症。在一些實施例中，癌症選自膀胱癌、乳癌、結腸直腸癌、腎癌、肺癌、淋巴瘤、黑色素瘤、口腔/口咽癌、胰臟癌、前列腺癌、甲狀腺癌及子宮癌。In some embodiments, the disease, disorder or condition is cancer. In some embodiments, the cancer is selected from bladder cancer, breast cancer, colorectal cancer, kidney cancer, lung cancer, lymphoma, melanoma, oral/oropharyngeal cancer, pancreatic cancer, prostate cancer, thyroid cancer and uterine cancer.

在一些實施例中，疾病、病症或疾患係遺傳病症。在一些實施例中，遺傳病症與功能獲得型突變或功能喪失型突變相關。In some embodiments, the disease, disorder, or condition is a genetic disorder. In some embodiments, the genetic disorder is associated with a gain-of-function mutation or a loss-of-function mutation.

在一些實施例中，疾病、病症或疾患係自體免疫疾病。在一些實施例中，自體免疫疾病選自青銅色病(Addison disease)、乳糜瀉、類風濕性關節炎、狼瘡、發炎性腸病、皮肌炎、多發性硬化、糖尿病、格林-巴利症候群(Guillain-Barre syndrome)、慢性發炎性去髓鞘型多發性神經病變、牛皮癬、惡性貧血、格雷氏病(graves’ disease)、橋本氏甲狀腺炎(Hashimoto’s thyroiditis)、重症肌無力及血管炎休格倫氏症候群(Sjogren syndrome)。In some embodiments, the disease, disorder, or condition is an autoimmune disease. In some embodiments, the autoimmune disease is selected from Addison disease, chylous diarrhea, rheumatoid arthritis, lupus, inflammatory bowel disease, dermatomyositis, multiple sclerosis, diabetes, Guillain-Barre syndrome, chronic inflammatory demyelinating polyneuropathy, psoriasis, pernicious anemia, Graves' disease, Hashimoto's thyroiditis, myasthenia gravis, and Sjogren's syndrome.

在一些實施例中，疾病、病症或疾患係罕見疾病。如本文所述，罕見疾病係指危及生命或慢性衰弱性疾病，其盛行率如此之低(例如，低於1/2000人)以致於需要特別的聯合努力來解決。In some embodiments, the disease, disorder, or condition is a rare disease. As used herein, a rare disease is a life-threatening or chronically debilitating disease that occurs so rarely (e.g., less than 1 in 2,000 people) that a special joint effort is required to address it.

在一些實施例中，本揭示案提供可選擇性靶向身體內之特定系統之複合物。如本文所用，提及「靶向」特定系統係指增加期望系統中衍生自複合物中之負荷之RNA的表現。舉例而言，在一些實施例中，本文所述之複合物可選擇性靶向肺、肝臟、脾、心臟、腦、淋巴結、膀胱、腎臟及胰臟。如本文所述，當單一靶表現之mRNA之量係投與後其他器官中之表現的65%或更多時，複合物「選擇性靶向」器官(例如，全身65%或更多之mRNA自單一器官表現，剩餘35%分佈在一或多個不同器官之間)。在一些實施例中，本文所述之複合物選擇性靶向肺。在一些實施例中，本文所述之複合物選擇性靶向肝臟。在一些實施例中，本文所述之複合物選擇性靶向脾。在一些實施例中，本文所述之複合物選擇性靶向心臟。In some embodiments, the present disclosure provides complexes that can selectively target specific systems in the body. As used herein, reference to "targeting" a specific system refers to increasing the expression of RNA derived from the cargo in the complex in the desired system. For example, in some embodiments, the complexes described herein can selectively target the lungs, liver, spleen, heart, brain, lymph nodes, bladder, kidneys, and pancreas. As described herein, a complex is "selectively targeted" to an organ when the amount of mRNA expressed by a single target is 65% or more of the expression in other organs after administration (e.g., 65% or more of the mRNA throughout the body is expressed from a single organ, with the remaining 35% distributed among one or more different organs). In some embodiments, the complexes described herein selectively target the lungs. In some embodiments, the complexes described herein selectively target the liver. In some embodiments, the complexes described herein are selectively targeted to the spleen. In some embodiments, the complexes described herein are selectively targeted to the heart.

遞送方法本揭示案尤其提供粒子，其納入組合物(例如，如本文所提及之醫藥組合物或醫藥調配物)中以投與個體。舉例而言，在一些實施例中，包含本文所述粒子之組合物係作為單一療法投與。在一些實施例中，包含本文所述粒子之組合物係作為組合療法的一部分投與。在一些實施例中，本文所述之組合物中總RNA之濃度(例如，所有一或多種RNA分子之總濃度)係約0.01 mg/mL至約1.0 mg/mL、或0.03 mg/mL至約0.3 mg/mL、或約0.05 mg/mL至約0.15 mg/mL。Delivery Methods The present disclosure provides, inter alia, particles that are incorporated into compositions (e.g., pharmaceutical compositions or pharmaceutical formulations as described herein) for administration to a subject. For example, in some embodiments, a composition comprising the particles described herein is administered as a single therapy. In some embodiments, a composition comprising the particles described herein is administered as part of a combination therapy. In some embodiments, the concentration of total RNA (e.g., the total concentration of all one or more RNA molecules) in a composition described herein is about 0.01 mg/mL to about 1.0 mg/mL, or 0.03 mg/mL to about 0.3 mg/mL, or about 0.05 mg/mL to about 0.15 mg/mL.

組合物(亦稱為醫藥組合物)可另外包含醫藥學上可接受之賦形劑，其如本文所用包括如適於期望具體劑量形式之任何及所有溶劑、分散介質、稀釋劑或其他液體媒劑、分散或懸浮助劑、表面活性劑、等滲劑、增稠或乳化劑、防腐劑、固體黏合劑、潤滑劑及諸如此類。Remington's The Science and Practice of Pharmacy，第21版，A. R. Gennaro (Lippincott, Williams & Wilkins, Baltimore, MD, 2006；其全文皆以引用方式併入本文中)揭示用於調配醫藥組合物之各種賦形劑及製備其之已知技術。除非任何習用賦形劑介質與物質或其衍生物不相容，例如藉由產生任何不期望生物效應或另外以有害方式與醫藥組合物之任何其他組分相互作用，否則其使用預期在本揭示案之範圍內。The composition (also referred to as a pharmaceutical composition) may further comprise a pharmaceutically acceptable excipient, which, as used herein, includes any and all solvents, dispersion media, diluents or other liquid vehicles, dispersing or suspending aids, surfactants, isotonic agents, thickening or emulsifying agents, preservatives, solid binders, lubricants, and the like, suitable for the desired specific dosage form. Remington's The Science and Practice of Pharmacy, 21st ed., A. R. Gennaro (Lippincott, Williams & Wilkins, Baltimore, MD, 2006; incorporated herein by reference in its entirety) discloses various excipients used to formulate pharmaceutical compositions and known techniques for preparing the same. Unless any customary excipient medium is incompatible with the substance or its derivatives, for example by producing any undesirable biological effect or otherwise interacting in a deleterious manner with any other component of the pharmaceutical composition, its use is contemplated to be within the scope of the present disclosure.

在一些實施例中，賦形劑經批准用於人類及用於獸醫用途。在一些實施例中，賦形劑係由美國食品藥品管理局(United States Food and Drug Administration)核准。在一些實施例中，賦形劑係醫藥級。在一些實施例中，賦形劑滿足美國藥典(United States Pharmacopoeia，USP)、歐洲藥典(European Pharmacopoeia，EP)、英國藥典(British Pharmacopoeia)及/或國際藥典之標準。In some embodiments, the excipient is approved for human and veterinary use. In some embodiments, the excipient is approved by the United States Food and Drug Administration. In some embodiments, the excipient is pharmaceutical grade. In some embodiments, the excipient meets the standards of the United States Pharmacopoeia (USP), the European Pharmacopoeia (EP), the British Pharmacopoeia, and/or the International Pharmacopoeia.

用於製造醫藥組合物之醫藥學上可接受之賦形劑包括但不限於惰性稀釋劑、分散劑及/或造粒劑、表面活性劑及/或乳化劑、崩解劑、黏合劑、防腐劑、緩衡劑、潤滑劑及/或油。該等賦形劑可視情況地包括在醫藥調配物中。根據調配者之判斷，諸如可可脂及栓劑蠟、著色劑、包衣劑、甜味劑、矯味劑及/或芳香劑之賦形劑可存在於組合物中。Pharmaceutically acceptable excipients used in the manufacture of pharmaceutical compositions include, but are not limited to, inert diluents, dispersants and/or granulating agents, surfactants and/or emulsifiers, disintegrants, binders, preservatives, balancing agents, lubricants, and/or oils. Such excipients may be included in the pharmaceutical formulation as appropriate. Excipients such as cocoa butter and suppository wax, coloring agents, coating agents, sweeteners, flavoring agents, and/or fragrances may be present in the composition at the discretion of the formulator.

醫藥劑之調配及/或製造中之一般考慮因素可參見例如Remington: The Science and Practice of Pharmacy，第21版，Lippincott Williams & Wilkins, 2005 (其全文皆以引用方式併入本文中)。General considerations in the formulation and/or manufacture of pharmaceuticals can be found, for example, in Remington: The Science and Practice of Pharmacy, 21st ed., Lippincott Williams & Wilkins, 2005 (incorporated herein by reference in its entirety).

在一些實施例中，本文所提供之醫藥組合物可根據習用技術(例如Remington: The Science and Practice of Pharmacy，第21版，Lippincott Williams & Wilkins, 2005 (其全文皆以引用方式併入本文中)中所揭示之技術)與一或多種醫藥學上可接受之載劑或稀釋劑以及任何其他已知佐劑及賦形劑一起調配。In some embodiments, the pharmaceutical compositions provided herein can be formulated according to customary techniques, such as those disclosed in Remington: The Science and Practice of Pharmacy, 21st edition, Lippincott Williams & Wilkins, 2005 (incorporated herein by reference in its entirety), together with one or more pharmaceutically acceptable carriers or diluents and any other known adjuvants and excipients.

本文所述之醫藥複合物及組合物可藉由此項技術中已知之適當方法投與。如熟習此項技術者應瞭解，投與之途徑及/或模式可端視多種因素而定，包括例如(但不限於)本文所述醫藥組合物之穩定性及/或藥物動力學及/或藥效學。The pharmaceutical compounds and compositions described herein can be administered by any suitable method known in the art. As will be appreciated by those skilled in the art, the route and/or mode of administration may depend on a variety of factors, including,for example , but not limited to, the stability and/or pharmacokinetic and/or pharmacodynamic properties of the pharmaceutical compositions described herein.

在一些實施例中，本文所述之醫藥組合物經調配用於非經腸投與，其包括除腸及局部投與外之投與模式，通常藉由注射，且包括(但不限於)靜脈內、肌內、動脈內、鞘內、囊內、眶內、心內、真皮內、腹膜內、經氣管、皮下、表皮下、關節內、囊下、蛛膜下、脊柱內、硬膜外及胸骨內注射及輸注。In some embodiments, the pharmaceutical compositions described herein are formulated for parenteral administration, which includes modes of administration other than enteral and topical administration, usually by injection, and includes, but is not limited to, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcutaneous, intraarticular, subcapsular, subarachnoid, intraspinal, epidural, and intrasternal injection and infusion.

在一些實施例中，本文所述之醫藥組合物經調配用於靜脈內投與。在一些實施例中，可用於靜脈內投與之醫藥學上可接受之載劑包括無菌水溶液或分散液及製備無菌可注射溶液或分散液之無菌粉末。In some embodiments, the pharmaceutical compositions described herein are formulated for intravenous administration. In some embodiments, pharmaceutically acceptable carriers for intravenous administration include sterile aqueous solutions or dispersions and sterile powders for preparing sterile injectable solutions or dispersions.

在一些具體實施例中，本文所述之醫藥組合物經調配用於皮下(s.c)投與。在一些具體實施例中，本文所述之醫藥組合物經調配用於肌內(i.m)投與。In some embodiments, the pharmaceutical compositions described herein are formulated for subcutaneous (s.c.) administration. In some embodiments, the pharmaceutical compositions described herein are formulated for intramuscular (i.m.) administration.

治療組合物在製造及儲存條件下通常必須係無菌且穩定的。組合物可調配為溶液、分散液、粉末(例如，凍乾粉末)、微乳液、脂質奈米粒子或適於高藥物濃度之其他有序結構。載劑可為含有例如水、乙醇、多元醇(例如甘油、丙二醇及液體聚乙二醇及諸如此類)及其適宜混合物之溶劑或分散介質。可例如藉由使用包衣(例如卵磷脂)、在分散液之情形下藉由維持所需粒徑及藉由使用表面活性劑來維持適當流動性。在許多情形下，組合物中較佳應包括等滲劑，例如糖、多元醇(例如甘露醇、山梨醇)、或氯化鈉。在一些實施例中，可注射組合物之延長吸收可藉由將延遲吸收之劑(例如單硬脂酸鹽及明膠)納入組合物中來達成。Therapeutic compositions generally must be sterile and stable under the conditions of manufacture and storage. The compositions can be formulated as solutions, dispersions, powders (e.g. , lyophilized powders), microemulsions, lipid nanoparticles, or other ordered structures suitable for high drug concentrations. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, a polyol (e.g., glycerol, propylene glycol, and liquid polyethylene glycol, and the like), and suitable mixtures thereof. Adequate fluidity can be maintained, for example, by the use of a coating (e.g., lecithin), by maintaining the desired particle size in the case of dispersions, and by the use of a surfactant. In many cases, it is preferable to include an isotonic agent, such as a sugar, a polyol (e.g., mannitol, sorbitol), or sodium chloride, in the composition. In some embodiments, prolonged absorption of the injectable compositions can be brought about by incorporating into the composition an agent that delays absorption, for example, monostearate and gelatin.

無菌可注射溶液可藉由將所需量之活性化合物與上文所列舉之一種成分或成分之組合(視需要)一起納入適當溶劑中、然後滅菌微濾來製備。Sterile injectable solutions can be prepared by incorporating the active compound in the required amount in an appropriate solvent with one or a combination of ingredients enumerated above, as required, followed by sterilization and microfiltration.

在一些實施例中，分散液係藉由將活性化合物納入含有基礎分散介質及來自上文所列舉成分之所需其他成分之無菌媒劑中來製備。在用於製備無菌可注射溶液之無菌粉末之情形下，較佳製備方法係真空乾燥及冷凍乾燥(凍乾)，其產生活性成分加來自其先前經無菌過濾溶液之任何其他期望成分的粉末。In some embodiments, dispersions are prepared by incorporating the active compound into a sterile vehicle containing a basic dispersion medium and the desired other ingredients from those listed above. In the case of sterile powders for the preparation of sterile injectable solutions, the preferred methods of preparation are vacuum drying and freeze drying (lyophilization), which yield a powder of the active ingredient plus any other desired ingredients from a previously sterile-filtered solution thereof.

可用於本文所述之醫藥組合物中之適宜水性及非水性載劑之實例包括水、乙醇、多元醇(例如甘油、丙二醇、聚乙二醇及諸如此類)以及其適宜混合物、植物油(例如橄欖油)及可注射有機酯(例如油酸乙酯)。可例如藉由使用包衣材料(例如卵磷脂)、在分散液之情形下藉由維持所需粒徑及藉由使用表面活性劑來維持適當流動性。Examples of suitable aqueous and non-aqueous carriers that can be used in the pharmaceutical compositions described herein include water, ethanol, polyols (e.g., glycerol, propylene glycol, polyethylene glycol, and the like), and suitable mixtures thereof, vegetable oils (e.g., olive oil), and injectable organic esters (e.g., ethyl oleate). Proper fluidity can be maintained, for example, by the use of coating materials (e.g., lecithin), by maintaining the required particle size in the case of dispersions, and by the use of surfactants.

該等組合物亦可含有佐劑，例如防腐劑、潤濕劑、乳化劑及分散劑。可藉由滅菌程式及藉由包括各種抗細菌劑及抗真菌劑，例如對羥基苯甲酸酯、氯丁醇、苯酚山梨酸及其類似物來確保預防微生物之存在。亦可期望將等滲劑(諸如糖、氯化鈉及其類似劑)包括至本文所述之醫藥組合物中。另外，可藉由包括延遲吸收之劑(諸如單硬脂酸鋁及明膠)來實現可注射醫藥形式之延長吸收。The compositions may also contain adjuvants such as preservatives, wetting agents, emulsifiers, and dispersants. Prevention of the presence of microorganisms can be ensured by sterilization procedures and by the inclusion of various antibacterial and antifungal agents, such as para-hydroxybenzoate, chlorobutanol, phenol, sorbic acid, and the like. It may also be desirable to include isotonic agents (such as sugars, sodium chloride, and the like) in the pharmaceutical compositions described herein. In addition, prolonged absorption of injectable pharmaceutical forms can be achieved by including agents that delay absorption (such as aluminum monostearate and gelatin).

本文所述醫藥組合物之調配物可藉由藥理學技術中已知或後來開發出之任一方法製備。一般而言，該等製備方法包括以下步驟：使活性成分與稀釋劑或另一賦形劑及/或一或多種其他輔助成分締合，且然後若必要及/或需要，將產物成型及/或包裝成所需單劑量或多劑量單位。The formulations of the pharmaceutical compositions described herein can be prepared by any method known or later developed in the art of pharmacology. Generally speaking, such preparation methods include the steps of combining the active ingredient with a diluent or another excipient and/or one or more other auxiliary ingredients, and then, if necessary and/or desired, shaping and/or packaging the product into the desired single or multiple dosage units.

本揭示案之醫藥組合物可以整體、以單一單位劑量、及/或以複數個單一單位劑量來製備、包裝及/或出售。如本文所用之「單位劑量」係包含預定量之使用本文所述之系統及/或方法產生之至少一種RNA產物的醫藥組合物之離散量。The pharmaceutical compositions of the present disclosure can be prepared, packaged, and/or sold in bulk, in a single unit dose, and/or in a plurality of single unit doses. As used herein, a "unit dose" is a discrete amount of a pharmaceutical composition comprising a predetermined amount of at least one RNA product produced using the systems and/or methods described herein.

在一些實施例中，可包括在本文所述之醫藥組合物中之活性劑係或包含在本文所述之組合療法中投與之治療劑。本文所述之醫藥組合物可在組合療法中投與，亦即與其他劑組合。在一些實施例中，該等治療劑可包括導致調節T細胞清除或功能性失活之劑。舉例而言，在一些實施例中，組合療法可包括所提供之醫藥組合物與至少一種免疫檢查點抑制劑。In some embodiments, the active agents included in the pharmaceutical compositions described herein are or are included in therapeutic agents administered in combination therapies described herein. The pharmaceutical compositions described herein can be administered in combination therapies, i.e., in combination with other agents. In some embodiments, such therapeutic agents can include agents that cause the elimination or functional inactivation of regulatory T cells. For example, in some embodiments, a combination therapy can include a provided pharmaceutical composition and at least one immune checkpoint inhibitor.

在一些實施例中，本文所述之醫藥組合物可與放射療法及/或自體外周幹細胞或骨髓移植結合投與。In some embodiments, the pharmaceutical compositions described herein can be administered in combination with radiation therapy and/or autologous peripheral stem cell or bone marrow transplantation.

在一些實施例中，本文所述之醫藥組合物可冷凍以允許長期儲存。In some embodiments, the pharmaceutical compositions described herein can be frozen to allow for long-term storage.

本揭示案之醫藥組合物可呈冷凍形式或呈「即用形式」(亦即呈可立即投與個體之形式，尤其液體形式，例如，無需任何處理，諸如解凍、重構或稀釋)。因此，在投與醫藥組合物之可儲存形式之前，必須將此可儲存形式加工或轉移成即用型或可投與形式。例如，必須解凍冷凍之醫藥組合物。即用型可注射劑可存在於諸如小瓶、安瓿或注射器之容器中，其中容器可含有一或多個劑量。The pharmaceutical compositions of the present disclosure may be in a frozen form or in a "ready-to-use form" (i.e., in a form, particularly a liquid form, that can be immediately administered to a subject,e.g. , without any processing, such as thawing, reconstitution, or dilution). Therefore, prior to administration of the storable form of the pharmaceutical composition, the storable form must be processed or converted into a ready-to-use or administrable form.For example , a frozen pharmaceutical composition must be thawed. Ready-to-use injectables may be in containers such as vials, ampoules, or syringes, wherein the container may contain one or more doses.

在一實施例中，將醫藥組合物凍乾。在一實施例中，將醫藥組合物噴霧乾燥。此等技術為熟習此項技術者所熟知。In one embodiment, the pharmaceutical composition is freeze-dried. In another embodiment, the pharmaceutical composition is spray-dried. These techniques are well known to those skilled in the art.

在一些實施例中，醫藥組合物呈冷凍形式且可在約-90℃或更高，諸如約-90℃至約-10℃之溫度下儲存。舉例而言，本文所述之冷凍醫藥組合物可在約-90℃至約-10℃，諸如約-90℃至約-40℃、或約-40℃至約-25℃、或約-25℃至約-10℃範圍內之溫度，或約-20℃之溫度下儲存。In some embodiments, the pharmaceutical composition is in a frozen form and can be stored at a temperature of about -90°C or higher, such as about -90°C to about -10°C. For example, the frozen pharmaceutical compositions described herein can be stored at a temperature in the range of about -90°C to about -10°C, such as about -90°C to about -40°C, or about -40°C to about -25°C, or about -25°C to about -10°C, or at a temperature of about -20°C.

在呈冷凍形式之醫藥組合物之一些實施例中，醫藥組合物可儲存至少1週，諸如至少2週、至少3週、至少4週、至少1個月、至少2個月、至少3個月、至少6個月、至少12個月、至少24個月或至少36個月，較佳至少4週。舉例而言，冷凍之醫藥組合物可在-20℃下儲存至少4週、較佳至少1個月、更佳至少2個月、更佳至少3個月、更佳至少6個月。In some embodiments of the pharmaceutical composition in frozen form, the pharmaceutical composition can be stored for at least 1 week, such as at least 2 weeks, at least 3 weeks, at least 4 weeks, at least 1 month, at least 2 months, at least 3 months, at least 6 months, at least 12 months, at least 24 months, or at least 36 months, preferably at least 4 weeks. For example, the frozen pharmaceutical composition can be stored at -20°C for at least 4 weeks, preferably at least 1 month, more preferably at least 2 months, more preferably at least 3 months, and more preferably at least 6 months.

在呈冷凍形式之醫藥組合物之一些實施例中，當核酸為mRNA時，解凍冷凍之醫藥組合物後之mRNA完整性為初始mRNA完整性之至少90%、至少95%、至少97%、至少98%或實質上100%，例如，在解凍已在-20℃下儲存(至少1週，諸如至少2週、至少3週、至少4週、至少1個月、至少2個月、至少3個月、至少6個月、至少12個月、至少24個月或至少36個月，較佳至少4週)之冷凍組合物後。In some embodiments of the pharmaceutical composition in frozen form, when the nucleic acid is mRNA, the mRNA integrity after thawing the frozen pharmaceutical composition is at least 90%, at least 95%, at least 97%, at least 98% or substantially 100% of the initial mRNA integrity,for example , after thawing a frozen composition that has been stored at -20°C for at least 1 week, such as at least 2 weeks, at least 3 weeks, at least 4 weeks, at least 1 month, at least 2 months, at least 3 months, at least 6 months, at least 12 months, at least 24 months, or at least 36 months, preferably at least 4 weeks.

在呈冷凍形式之醫藥組合物之一些實施例中，在解凍冷凍之醫藥組合物之後，粒子之大小(Z_平均值)及/或大小分佈及/或PDI基本上等於冷凍前初始醫藥組合物之粒子的大小(Z_平均值)及/或大小分佈及/或PDI。舉例而言，若自如本文所述之冷凍醫藥組合物製備即用型醫藥組合物，則較佳的係，即用型醫藥組合物中含有之粒子之大小(Z_平均值)和/或大小分佈和/或PDI基本上等於冷凍前冷凍醫藥組合物中含有之粒子的初始大小(Z_平均值)和/或大小分佈和/或PDI。In some embodiments of the pharmaceutical composition in frozen form, after thawing the frozen pharmaceutical composition, the size (Z_-average ) and/or size distribution and/or PDI of the particles are substantially equal to the size (Z-average) and/or size distribution and/or PDI of the particles of the initial pharmaceutical composition before freezing. For example, if a ready-to-use pharmaceutical composition is prepared from a frozen pharmaceutical composition as described herein, it is preferred that the size (Z_-average ) and/or size distribution and/or PDI of the particles contained in the ready-to-use pharmaceutical composition are substantially equal to the initial size (Z_-average ) and/or size distribution and/or PDI of the particles contained in the frozen pharmaceutical composition before freezing.

在一些實施例中，當核酸為mRNA時，在一個冷凍/解凍循環後，較佳地在兩個冷凍/解凍循環後，更佳地在三個冷凍/解凍循環後，更佳地在四個冷凍/解凍循環之後，更佳地在五個冷凍/解凍循環或更多次之後，醫藥組合物之mRNA粒子之大小及mRNA完整性基本上等於初始醫藥組合物之mRNA粒子之大小及mRNA完整性(亦即，在醫藥組合物第一次冷凍之前)。In some embodiments, when the nucleic acid is mRNA, after one freeze/thaw cycle, preferably after two freeze/thaw cycles, more preferably after three freeze/thaw cycles, more preferably after four freeze/thaw cycles, and more preferably after five freeze/thaw cycles or more, the size of the mRNA particles and the mRNA integrity of the pharmaceutical composition are substantially equivalent to the size of the mRNA particles and the mRNA integrity of the initial pharmaceutical composition (i.e. , before the first freezing of the pharmaceutical composition).

在一些實施例中，醫藥組合物呈液體形式且可在約0℃至約20℃範圍內之溫度下儲存。舉例而言，本文所述之液體醫藥組合物可在約1℃至約15℃，諸如約2℃至約10℃、或約2℃至約8℃範圍內之溫度下，或在約5℃之溫度下儲存。In some embodiments, the pharmaceutical composition is in liquid form and can be stored at a temperature in the range of about 0° C. to about 20° C. For example, the liquid pharmaceutical compositions described herein can be stored at a temperature in the range of about 1° C. to about 15° C., such as about 2° C. to about 10° C., or about 2° C. to about 8° C., or at a temperature of about 5° C.

在一些實施例中，當核酸為mRNA時，醫藥組合物在儲存時之mRNA完整性為初始mRNA完整性(亦即，初始醫藥組合物之mRNA完整性)之至少70%，較佳至少80%，更佳至少90%。In some embodiments, when the nucleic acid is mRNA, the mRNA integrity of the pharmaceutical composition upon storage is at least 70%, preferably at least 80%, and more preferably at least 90% of the initial mRNA integrity (i.e., the mRNA integrity of the initial pharmaceutical composition).

在呈液體形式之醫藥組合物之一些實施例中，醫藥組合物可儲存至少1週，諸如至少2週、至少3週、至少4週、至少1個月、至少2個月、至少3個月、至少6個月、至少12個月或至少24個月，較佳至少4週。舉例而言，液體醫藥組合物可在5℃下儲存至少4週、較佳至少1個月、更佳至少2個月、更佳至少3個月、更佳至少6個月。In some embodiments of the pharmaceutical composition in liquid form, the pharmaceutical composition can be stored for at least 1 week, such as at least 2 weeks, at least 3 weeks, at least 4 weeks, at least 1 month, at least 2 months, at least 3 months, at least 6 months, at least 12 months, or at least 24 months, preferably at least 4 weeks. For example, the liquid pharmaceutical composition can be stored at 5° C. for at least 4 weeks, preferably at least 1 month, more preferably at least 2 months, more preferably at least 3 months, and more preferably at least 6 months.

在呈液體形式之醫藥組合物之一些實施例中，當核酸為mRNA時，當在例如0℃或更高溫度下儲存至少一週時，液體組合物之mRNA完整性使得可達成期望效應，例如，誘導免疫反應。舉例而言，當在例如在0℃或更高溫度下儲存至少一週(諸如至少2週、至少三週、至少四週、至少一個月、至少兩個月、至少三個月、至少4個月或至少6個月)時，液體組合物之mRNA完整性與初始組合物之mRNA完整性(亦即，組合物儲存前之mRNA完整性)相比可為至少90%。在一些實施例中，在儲存至少四週(例如至少三個月)後，較佳地在0℃或更高，諸如約2℃至約8℃之溫度下，組合物的mRNA完整性與儲存前之mRNA完整性相比為至少90%。In some embodiments of the pharmaceutical composition in liquid form, when the nucleic acid is mRNA, the mRNA integrity of the liquid composition is such that a desired effect,e.g. , induction of an immune response, can be achieved when stored, e.g., at 0° C. or higher for at least one week. For example, the mRNA integrity of the liquid composition can be at least 90% when stored, e.g., at 0° C. or higher for at least one week (e.g., at least two weeks, at least three weeks, at least four weeks, at least one month, at least two months, at least three months, at least four months, or at least six months) compared to the mRNA integrity of the initial composition (i.e. , the mRNA integrity of the composition before storage). In some embodiments, after storage for at least four weeks (e.g., at least three months), preferably at 0°C or higher, such as about 2°C to about 8°C, the mRNA integrity of the composition is at least 90% compared to the mRNA integrity before storage.

在一些實施例中，當核酸為mRNA時，醫藥組合物之初始mRNA完整性(亦即，在其製備之後但儲存之前)為至少50%，且在儲存至少一週(諸如至少2週、至少三週、至少四週、至少一個月、至少兩個月或至少3個月)後，較佳地在0℃或更高，諸如約2℃至約8℃之溫度下，醫藥組合物之mRNA完整性為初始mRNA完整性之至少90%。In some embodiments, when the nucleic acid is mRNA, the initial mRNA integrity of the pharmaceutical composition (i.e. , after its preparation but before storage) is at least 50%, and after storage for at least one week (e.g., at least 2 weeks, at least three weeks, at least four weeks, at least one month, at least two months, or at least 3 months), preferably at a temperature of 0°C or higher, such as about 2°C to about 8°C, the mRNA integrity of the pharmaceutical composition is at least 90% of the initial mRNA integrity.

在呈液體形式之醫藥組合物之一些實施例中，當在例如0℃或更高溫度下儲存至少一週時，醫藥組合物之粒子之大小(Z_平均值) (及/或大小分佈及/或多分散性指數(PDI))使得可達成期望效應，例如，誘導免疫反應。舉例而言，當在例如0℃或更高溫度下儲存至少一週時，醫藥組合物之粒子之大小(Z_平均值) (及/或大小分佈及/或多分散性指數(PDI))基本上等於初始醫藥組合物(亦即，儲存前)之粒子之大小(Z_平均值) (及/或大小分佈及/或PDI)。In some embodiments of the pharmaceutical composition in liquid form, when stored at, for example, 0° C. or higher for at least one week, the size (Z_-average ) (and/or size distribution and/or polydispersity index (PDI)) of the particles of the pharmaceutical composition is such that a desired effect, for example, induction of an immune response, can be achieved. For example, when stored at, for example, 0° C. or higher for at least one week, the size (Z_-average ) (and/or size distribution and/or polydispersity index (PDI)) of the particles of the pharmaceutical composition is substantially equal to the size (Z_-average ) (and/or size distribution and/or PDI) of the particles of the initial pharmaceutical composition (i.e. , before storage).

在一些實施例中，當在例如0℃或更高溫度下儲存至少一週時，醫藥組合物之粒子之大小(Z_平均值)介於約50 nm與約500 nm之間，較佳介於約40 nm與約200 nm之間，更佳介於約40 nm與約120 nm之間。在一些實施例中，當在例如0℃或更高溫度下儲存至少一週時，醫藥組合物之粒子之PDI小於0.3，較佳地小於0.2，更佳地小於0.1。In some embodiments, when stored at,for example, 0° C. or higher for at least one week, the size (Z_-average ) of the particles of the pharmaceutical composition is between about 50 nm and about 500 nm, preferably between about 40 nm and about 200 nm, and more preferably between about 40 nm and about 120 nm. In some embodiments, when stored at,for example, 0° C. or higher for at least one week, the PDI of the particles of the pharmaceutical composition is less than 0.3, preferably less than 0.2, and more preferably less than 0.1.

在一些實施例中，例如在0℃或更高溫度下儲存至少一週後，醫藥組合物之粒子之大小(Z_平均值)介於約50 nm與約500 nm之間，較佳介於約40 nm與約200 nm，更佳介於約40 nm與約120 nm之間，且例如在0℃或更高溫度下儲存至少一週後，醫藥組合物之粒子之大小(Z_平均值) (及/或大小分佈及/或PDI)基本上等於儲存前粒子之大小(Z_平均值) (及/或大小分佈及/或PDI)。在一些實施例中，例如在0℃或更高溫度下儲存至少一週後，醫藥組合物之粒子之大小(Z_平均值)介於約50 nm與約500 nm之間，較佳介於約40 nm與約200 nm，更佳介於約40 nm與約120 nm之間，且例如在0℃或更高溫度下儲存至少一週後，醫藥組合物之粒子之PDI小於0.3 (較佳地小於0.2，更佳地小於0.1)。In some embodiments, the size (Z_-average ) of the particles of the pharmaceutical composition is between about 50 nm and about 500 nm,preferably between about 40 nm and about 200 nm, and more preferably between about 40 nm and about 120 nm,for example after storage at 0° C. or higher for at least one week, and the size (Z-_{average) (and/or size distribution and/or PDI) of the particles of the pharmaceutical composition is substantially the same as the size (Z-average} ) (and/or size distribution and/or PDI) of the particles before storage, for example after storage at₀ ° C. or higher for at least one week. In some embodiments, the size (Z_-average ) of the particles of the pharmaceutical composition is between about 50 nm and about 500 nm, preferably between about 40 nm and about 200 nm, more preferably between about 40 nm and about 120 nm,forexample after storage at 0° C. or higher for at least one week, and the PDI of the particles of the pharmaceutical composition is less than 0.3 (preferably less than 0.2, more preferably less than 0.1), for example after storage at 0° C. or higher for at least one week.

儘管本文所提供之醫藥組合物之描述主要涉及適用於向人類投與的醫藥組合物，但熟習此項技術者將理解，此類組合物一般適用於向各種動物投與。為使適於投與人類之醫藥組合物適於投與各種動物而對該等組合物進行之修飾為此項技術中所熟知，且普通熟練獸醫藥理學家可僅用普通(若有)實驗設計及/或進行此類修飾。Although the descriptions of pharmaceutical compositions provided herein primarily relate to pharmaceutical compositions suitable for administration to humans, those skilled in the art will understand that such compositions are generally suitable for administration to various animals. Modifications of pharmaceutical compositions suitable for administration to humans to make them suitable for administration to various animals are well known in the art, and such modifications can be performed by an ordinarily skilled veterinary pharmacist using only ordinary (if any) experimental design and/or practice.

用於製備脂質之方法如本文所述，本揭示案提供一種製備陽離子或可離子化脂質(諸如本文所述之彼等脂質)之方法。製備本文所述之陽離子或可離子化脂質之方法不僅限於具有對稱脂族尾之彼等方法，且可應用於多種脂質結構。Methods for Preparing Lipids As described herein, the present disclosure provides a method for preparing cationic or ionizable lipids, such as those described herein. The methods for preparing cationic or ionizable lipids described herein are not limited to those having symmetrical aliphatic tails and can be applied to a variety of lipid structures.

舉例而言，在一些實施例中，本揭示案提供製備式III化合物：III 或其醫藥學上可接受之鹽之方法，該方法包含使式IV-1化合物與式IV-2化合物在熱存在下接觸：以提供第一中間產物，且其中使該第一中間產物與鉑觸媒及第一酸在氫氣存在下接觸以提供第二中間產物，且其中使該第二中間產物與第二酸接觸以提供該式III化合物，其中 M^1’和M^2’獨立地選自鍵或視情況經取代之C₂-C₁₀脂族基； L^1'及L^2'各自獨立地選自鍵、-OC(O)-及-C(O)O-； T^1’及T^2’各自獨立地選自視情況經取代之C₂-C₂₀脂族基； L^3’為視情況經取代之C₁-C₆脂族基； LG¹為適宜之脫離基；且 PG為適宜之醇保護基。For example, in some embodiments, the present disclosure provides methods for preparing compounds of formula III: III or a pharmaceutically acceptable salt thereof, which comprises contacting a compound of formula IV-1 with a compound of formula IV-2 in the presence of heat: to provide a first intermediate product, and wherein the first intermediate product is contacted with a platinum catalyst and a first acid in the presence of hydrogen to provide a second intermediate product, and wherein the second intermediate product is contacted with a second acid to provide the compound of formula III, wherein M^1' and M^2' are independently selected from a bond or an optionally substituted C₂ -C₁₀ aliphatic group; L^{1 '} and L^{2 '} are each independently selected from a bond, -OC(O)- and -C(O)O-; T^1' and T^2' are each independently selected from an optionally substituted C₂ -C₂₀ aliphatic group; L^3' is an optionally substituted C₁ -C₆ aliphatic group; LG¹ is a suitable leaving group; and PG is a suitable alcohol protecting group.

在一些實施例中，式IV-2化合物藉由以下方式製備：使式V-1a及式V-1b化合物與式V-2化合物在鈀偶合觸媒、銅(I)輔觸媒及胺鹼存在下接觸，以提供該式IV-2化合物，其中LG²為適宜之脫離基。In some embodiments, the compound of formula IV-2 is prepared by reacting the compounds of formula V-1a and formula V-1b with the compound of formula V-2. The reaction is carried out in the presence of a palladium coupling catalyst, a copper (I) cocatalyst and an amine base to provide the compound of formula IV-2, wherein LG² is a suitable detached group.

在一些實施例中，本揭示案提供一種製備式III-1化合物，III-1 或其醫藥學上可接受之鹽之方法，該方法包含使式IV-1化合物與式IV-3化合物在熱存在下接觸：以提供第一中間產物，且其中使該第一中間產物與酸接觸，以提供該式III-1化合物，其中 M^1’和M^2’各自獨立地選自鍵或視情況經取代之C₂-C₁₀脂族基； T^1’及T^2’各自獨立地選自視情況經取代之C₂-C₂₀脂族基； L^3’為視情況經取代之C₁-C₆脂族基； LG¹為適宜之脫離基；且 PG為適宜之醇保護基。In some embodiments, the present disclosure provides a method for preparing a compound of formula III-1, III-1 or a pharmaceutically acceptable salt thereof, the method comprising contacting a compound of formula IV-1 with a compound of formula IV-3 in the presence of heat: to provide a first intermediate product, and wherein the first intermediate product is contacted with an acid to provide the compound of formula III-1, wherein M^1' and M^2' are each independently selected from a bond or an optionally substituted C₂ -C₁₀ aliphatic group; T^1' and T^2' are each independently selected from an optionally substituted C₂ -C₂₀ aliphatic group; L^3' is an optionally substituted C₁ -C₆ aliphatic group; LG¹ is a suitable leaving group; and PG is a suitable alcohol protecting group.

在一些實施例中，式IV-3化合物藉由以下方式製備：使式IV-4化合物：IV-4 與一或多個當量之式IV-5及IV-6化合物中之每一者：在4-(二甲基胺基)吡啶(DMAP)及1-乙基-3-(3-二甲基胺基丙基)碳二亞胺(EDCl)存在下接觸，以提供該式IV-3化合物。In some embodiments, the compound of formula IV-3 is prepared by: making a compound of formula IV-4: IV-4 and one or more equivalents of each of the compounds of formula IV-5 and IV-6: The reaction mixture is contacted in the presence of 4-(dimethylamino)pyridine (DMAP) and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDCl) to provide the compound of formula IV-3.

在一些實施例中，式IV-4化合物藉由以下方式製備：使式IV-7化合物IV-7 與氫氣在鉑或鈀還原觸媒存在下接觸，以提供該式IV-4化合物。In some embodiments, the compound of formula IV-4 is prepared by: IV-7 is contacted with hydrogen in the presence of a platinum or palladium reducing catalyst to provide the compound of formula IV-4.

在一些實施例中，式IV-7化合物藉由以下方式製備：使V-2化合物與一個當量之式V-1c及V-1d化合物中之每一者在鈀偶合觸媒、銅(I)輔觸媒及胺鹼存在下接觸，以提供該式IV-2化合物，其中LG²為適宜之脫離基。In some embodiments, the compound of formula IV-7 is prepared by reacting the compound of formula V-2 with one equivalent of each of the compounds of formula V-1c and V-1d. The reaction is carried out in the presence of a palladium coupling catalyst, a copper (I) cocatalyst and an amine base to provide the compound of formula IV-2, wherein LG² is a suitable detached group.

如本文所述，M^1’和M^2’獨立地選自鍵或視情況經取代之C₂-C₁₀脂族基。在一些實施例中，M^1’及M^2’係相同的。在一些實施例中，M^1’及M^2’係不同的。在一些實施例中，M^1'及M^2'各自獨立地選自視情況經取代之C₂-C₁₀伸烷基。在一些實施例中，M^1’及M^2’各自獨立地選自–(CH₂)_4-6-。As described herein, M^1' and M^2' are independently selected from a bond or an optionally substituted C₂ -C₁₀ aliphatic group. In some embodiments, M^1' and M^2' are the same. In some embodiments, M^1' and M^2' are different. In some embodiments, M^{1 '} and M^{2 '} are each independently selected from an optionally substituted C₂ -C₁₀ alkylene group. In some embodiments, M^1' and M^2' are each independently selected from -(CH₂ )_4-6 -.

如本文所述，L^1'及L^2'各自獨立地選自鍵、-OC(O)-及-C(O)O-。在一些實施例中，L^1’及L^2’係相同的。在一些實施例中，L^1’及L^2’係不同的。在一些實施例中，L^1’係鍵。在一些實施例中，L^1’係-OC(O)-。在一些實施例中，L^1’係-C(O)O-。在一些實施例中，L^2’係鍵。在一些實施例中，L^2’係-OC(O)-。在一些實施例中，L^2’係-C(O)O-。在一些實施例中，L^1’及L^2’各自係鍵。在一些實施例中，L^1’及L^2’各自係-OC(O)-。在一些實施例中，L^1’及L^2’各自係-C(O)O-。As described herein, L1^′ and L2^′ are each independently selected from a bond, -OC(O)-, and -C(O)O-. In some embodiments,^L1′ and^L2′ are the same. In some embodiments,^L1′ and^L2′ are different. In some embodiments,^L1′ is a bond. In some embodiments,^L1′ is -OC(O)-. In some embodiments,^L1′ is -C(O)O-. In some embodiments,^L2′ is a bond. In some embodiments,^L2′ is -OC(O)-. In some embodiments,^L2′ is -C(O)O-. In some embodiments,^L1′ and^L2′ are each a bond. In some embodiments, L^1′ and L^2′ are each —OC(O)—. In some embodiments, L^1′ and L^2′ are each —C(O)O—.

如本文所述，L^3'為視情況經取代之_C1-C6脂族基。在一些實施例中，L^3'為視情況經取代之C₃-C₅脂族基。在一些實施例中，L^3'為視情況經取代之C₁-C₆伸烷基。在一些實施例中，L^3'為C₃-C₅伸烷基。在一些實施例中，L^3'為-(CH₂)_1-6-。在一些實施例中，L^3'為-(CH₂)₃-、-(CH₂)₄-或-(CH₂)₅-。在一些實施例中，L^3'為-(CH₂)₃-。在一些實施例中，L^3'為-(CH₂)₄-。在一些實施例中，L^3'為-(CH₂)₅-。As described herein, L^3' is an optionally substituted_C1-C6 aliphatic group. In some embodiments, L^3' is an optionally substituted_C3 -_C5 aliphatic group. In some embodiments, L^3' is an optionally substituted_C1 -_C6 alkylene group. In some embodiments, L^3' is a_C3 -_C5 alkylene group. In some embodiments, L^3' is -(_CH2 )_1-6- . In some embodiments, L^3' is -(_CH2 )_3- , -(_CH2 )_4- , or -(_CH2 )_5- . In some embodiments, L^3' is -(_CH2 )_3- . In some embodiments, L^3' is -(_CH2 )_4- . In some embodiments, L^3' is -(CH₂ )₅ -.

如本文所述，T^1’及T^2’各自獨立地選自視情況經取代之C₂-C₂₀脂族基。在一些實施例中，T^1’及T^2’係相同的。在一些實施例中，T^1’及T^2’係不同的。在一些實施例中，T^1'及T^2'各自為分枝C₃-C₂₀脂族基。在一些實施例中，T^1'及T^2'各自為直鏈C₂-C₂₀脂族基。在一些實施例中，T^1'及T^2'中之一者為分枝C₃-C₂₀脂族基，且T^1'及T^2'中之另一者為直鏈C₂-C₂₀脂族基。As described herein,^T1' and^T2' are each independently selected from an optionally substituted_C2 -_C20 aliphatic group. In some embodiments,^T1' and^T2' are the same. In some embodiments,^T1' and^T2' are different. In some embodiments, T1^' and T2^' are each a branched_C3 -_C20 aliphatic group. In some embodiments, T1^' and T2^' are each a linear_C2 -_C20 aliphatic group. In some embodiments, one of T1^' and T2^' is a branched_C3 -_C20 aliphatic group, and the other of T1^' and T2^' is a linear_C2 -_C20 aliphatic group.

在一些實施例中，T^1’及T^2’各自獨立地選自、、、、、、、、、及。In some embodiments, T^1' and T^2' are each independently selected from 、 、 、 、 、 、 、 、 、 and .

在一些實施例中，-L^1’-T^1’選自：及， 且-L^2'-T^2'選自：及其中： 各n1獨立地為0、1、2、3、4、5或6；且 各n2獨立地為1、2、3、4、5、6、7、8、9或10。In some embodiments, -L^1' -T^1' is selected from: and , and -L^2' -T^2' is selected from: and wherein: each n1 is independently 0, 1, 2, 3, 4, 5 or 6; and each n2 is independently 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10.

如本文所述，LG¹為適宜之脫離基。在一些實施例中，LG¹為鹵素或-OTf。在一些實施例中，LG¹選自溴、氯或碘。在一些實施例中，LG¹為溴。As described herein, LG¹ is a suitable ionizing group. In some embodiments, LG¹ is a halogen or -OTf. In some embodiments, LG¹ is selected from bromine, chlorine, or iodine. In some embodiments, LG¹ is bromine.

如本文所述，PG為適宜之醇保護基。熟習此項技術者將容易地瞭解醇部分之適宜保護基。在一些實施例中，PG為乙醯基、苯甲醯基、芐基、甲氧基乙氧基甲基醚(MEM)基團、二甲氧基三苯甲基(DMT)、甲氧基甲基醚(MOM)基團、甲氧基三苯甲基(MMT)、對甲氧基芐基醚(PMB)基團、特戊醯基(Piv)基團、第三丁基醚基團、四氫吡喃基、四氫呋喃基團、三苯甲基或矽基醚，包括第三丁基二甲基矽基醚(TBS)。在一些實施例中，PG為TBS基團。As described herein, PG is a suitable alcohol protecting group. Suitable protecting groups for alcohol moieties will be readily apparent to those skilled in the art. In some embodiments, PG is an acetyl group, a benzyl group, a benzyl group, a methoxyethoxymethyl ether (MEM) group, a dimethoxytrityl group (DMT), a methoxymethyl ether (MOM) group, a methoxytrityl group (MMT),a p-methoxybenzyl ether (PMB) group, a p-pentanoyl (Piv) group, a t-butyl ether group, a tetrahydropyranyl group, a tetrahydrofuranyl group, a trityl group, or a silyl ether, including t-butyldimethylsilyl ether (TBS). In some embodiments, PG is a TBS group.

如本文所述，LG²為適宜之脫離基。在一些實施例中，各LG²獨立地選自鹵素及-OTf。在一些實施例中，各LG²獨立地選自溴、氯或碘。在一些實施例中，各LG²為溴。As described herein,^LG is a suitable ionizing group. In some embodiments, each^LG is independently selected from a halogen and -OTf. In some embodiments, each^LG is independently selected from bromine, chlorine, or iodine. In some embodiments,^each LG is bromine.

在一些實施例中，鉑觸媒為PtO₂。In some embodiments, the platinum catalyst is PtO₂ .

在一些實施例中，鈀還原觸媒為碳載鈀。In some embodiments, the palladium reduction catalyst is carbon-supported palladium.

在一些實施例中，第一酸為有機酸。在一些實施例中，有機酸為乙酸。In some embodiments, the first acid is an organic acid. In some embodiments, the organic acid is acetic acid.

在一些實施例中，第二酸為鹽酸。In some embodiments, the second acid is hydrochloric acid.

在一些實施例中，鈀偶合觸媒為Pd(PPh₃)₄。In some embodiments, the palladium coupling catalyst is Pd(PPh₃ )₄ .

在一些實施例中，銅(I)輔觸媒為CuI。In some embodiments, the copper (I) co-catalyst is CuI.

在一些實施例中，胺鹼為三乙胺。In some embodiments, the amine base is triethylamine.

在一些實施例中，本文所述之化合物(例如式I化合物)係根據本文所述之方法製備。In some embodiments, the compounds described herein (e.g., compounds of Formula I) are prepared according to the methods described herein.

示範性實施例儘管具有非限制性，以下經編號實施例係本揭示案之某些態樣之示範： 1. 一種式I化合物，I 或其醫藥學上可接受之鹽，其中 M¹及M²各自為-(CH₂)_p1-； L¹及L²各自選自鍵、-OC(O)-及-C(O)O-； T¹及T²各自為：； L³為視情況經取代之C₁-C₆脂族基； G¹為-OH或-N(R^a)₂，且 R^a為H、視情況經取代之C₁-C₆脂族基或視情況經取代之C₃-C₆環脂族基； p1為選自2-20之整數，包括端值； p2及p3各自為選自1-20之整數，包括端值，且p2及p3係相同的。 2. 如實施例1之化合物，其中p1為選自2-10之整數，包括端值。 3. 如實施例1或2之化合物，其中M¹為-(CH₂)_4-6-且M²為-(CH₂)_4-6-。 4. 如實施例1至3中任一項之化合物，其中M¹及M²係相同的。 5. 如實施例1至4中任一項之化合物，其中T¹及T²係相同的。 6. 如實施例1至5中任一項之化合物，其中L¹及L²係相同的。 7. 如實施例1至6中任一項之化合物，其中p2及p3各自為選自3、4、5、6、7、8或9之整數。 8. 如實施例7之化合物，其中p2及p3各自為選自4、5或6之整數。 9. 如實施例1之化合物，其中T¹及T²各自選自：、、、、及。 10. 如實施例1之化合物，其中M¹-L¹-T¹由以下表示：其中p2及p3各自為4、5或6。 11. 如實施例1或10之化合物，其中M²-L²-T²由以下表示：其中p2及p3各自為4、5或6。 12. 如實施例1之化合物，其中M¹-L¹-T¹及M²-L²-T²各自選自:、、、、、、、、、、、、、、、、及。 13. 如實施例1至12中任一項之化合物，其中-L³-G¹為-(CH₂)₃-OH或-(CH₂)₄-OH。 14. 如實施例1之化合物，其中該化合物由式II-1表示：II-1 其中*表示立體純碳原子。 15. 如實施例1至14中任一項之化合物，其中該化合物之特徵在於具有約15,000與30,000之間的維納路徑及-15與0之間的MMFF94能量。 16. 如實施例1至15中任一項之化合物，其中該化合物之特徵在於具有0.1與0.2之間的k。 17. 如實施例1之化合物，其中該化合物選自表1。 18. 一種醫藥組合物，其包含如實施例1至17中任一項之化合物及核酸。 19. 如實施例18之醫藥組合物，其進一步包含以下中之一或多者：中性脂質、聚合物偶聯脂質、類固醇或陰離子兩親物。 20. 如實施例19之醫藥組合物，其進一步包含中性脂質。 21. 如實施例19或20之醫藥組合物，其中該中性脂質為磷脂。 22. 如實施例21之醫藥組合物，其中該磷脂係或包含二硬脂醯基磷脂醯膽鹼(DSPC)、二油醯基磷脂醯膽鹼(DOPC)、二肉豆蔻醯基磷脂醯膽鹼(DMPC)、雙十五醯基磷脂醯膽鹼、二月桂醯基磷脂醯膽鹼、二棕櫚醯基磷脂醯膽鹼(DPPC)、二花生醯基磷脂醯膽鹼(DAPC)、二山崳醯基磷脂醯膽鹼(DBPC)、雙二十三醯基磷脂醯膽鹼(DTPC)、雙二十四醯基磷脂醯膽鹼(DLPC)、棕櫚醯基油醯基-磷脂醯膽鹼(POPC)、1,2-二-O-十八烯基-sn-甘油-3-磷酸膽鹼(18:0二醚PC)、1-油醯基-2-膽固醇基半琥珀醯基-sn-甘油-3-磷酸膽鹼(OChemsPC)、1-十六基-sn-甘油-3-磷酸膽鹼(C16 Lyso PC)、二油醯基磷脂醯乙醇胺(DOPE)、二硬脂醯基-磷脂醯乙醇胺(DSPE)、二棕櫚醯基-磷脂醯乙醇胺(DPPE)、二肉豆蔻醯基-磷脂醯乙醇胺(DMPE)、二月桂醯基-磷脂醯乙醇胺(DLPE)、二植醯基-磷脂醯乙醇胺(DPyPE)、1,2-二-(9Z-十八碳烯醯基)-sn-甘油-3-磷酸膽鹼(DOPG)、1,2-二棕櫚醯基-sn-甘油-3-磷酸-(1′-rac-甘油) (DPPG)、1-棕櫚醯基-2-油醯基-sn-甘油-3-磷酸乙醇胺(POPE)或N-棕櫚醯基-D-赤-鞘胺醯基磷酸膽鹼(SM)。 23. 如實施例19至22中任一項之醫藥組合物，其進一步包含聚合物偶聯脂質。 24. 如實施例23之醫藥組合物，其中該聚合物偶聯脂質為包含聚合物之脂質，該聚合物為聚乙二醇。 25. 如實施例24之醫藥組合物，其中該聚合物偶聯脂質選自PEG-DAG、PEG-PE、PEG-S-DAG、PEG2000-DMG、PEG-S-DMG、PEG-cer、PEG二烷氧基丙基胺基甲酸酯(例如，ω-甲氧基(聚乙氧基)乙基-N-(2,3-二(十四烷氧基)丙基)胺基甲酸酯或2,3-二(十四烷氧基)丙基-N-(ω-甲氧基(聚乙氧基)乙基)胺基甲酸酯)及其組合。 26. 如實施例19至25中任一項之醫藥組合物，其進一步包含類固醇。 27. 如實施例26之醫藥組合物，其中該類固醇為固醇。 28. 如實施例27之醫藥組合物，其中該固醇為膽固醇。 29. 如實施例19至28中任一項之醫藥組合物，其進一步包含陰離子兩親物。 30. 如實施例29之醫藥組合物，其中該陰離子兩親物選自DMGS、DPGS及DMGS。 31. 如實施例19至22及26至29中任一項之醫藥組合物，其中該醫藥組合物包含中性脂質、類固醇及陰離子兩親物，且該醫藥組合物不包括聚合物偶聯脂質。 32. 如實施例18至31中任一項之醫藥組合物，其中該核酸為RNA。 33. 如實施例32之醫藥組合物，其中該RNA為modRNA、saRNA、taRNA或uRNA。 34. 如實施例18至31中任一項之醫藥組合物，其中該核酸為DNA。 35. 如實施例18至31中任一項之醫藥組合物，其中該核酸為RNA及DNA之混合物。 36. 如實施例19至35中任一項之醫藥組合物，其中該醫藥組合物包含： 約30 mol%至約60 mol%之該化合物； 約20 mol%至約60 mol%之該類固醇； 約1 mol%至約2 mol%之該聚合物偶聯脂質；及 約5 mol%至約20 mol%之該中性脂質。 37. 如實施例19至35中任一項之醫藥組合物，其中該醫藥組合物包含： 約30 mol%至約60 mol%之該化合物； 約20 mol%至約60 mol%之該類固醇； 約0.5至約10 mol%之該陰離子兩親物；及 約5 mol%至約20 mol%之該中性脂質。 38. 如實施例37之醫藥組合物，其中該醫藥組合物不包括聚合物偶聯脂質。 39. 一種懸浮液，其包含分散相及水相，且其中該分散相包含呈粒子形式之如實施例18至38中任一項之醫藥組合物。 40. 如實施例39之懸浮液，其中該等粒子之平均直徑介於約50 nm至約200 nm之間。 41. 如實施例40之懸浮液，其中該水相包含該懸浮液中核酸之總濃度的小於20%。 42. 如實施例39至41中任一項之懸浮液，其中該水相實質上不含該核酸。 43. 一種治療疾病、病症或疾患之方法，其包含向個體投與如實施例39至42中任一項之懸浮液。 44. 如實施例43之方法，其中該疾病、病症或疾患選自傳染病、癌症、遺傳病症、自體免疫疾病或罕見疾病。 45. 一種增加個體之標靶中RNA表現或引起其增加之方法，其包含向該個體投與如實施例39至42中任一項之懸浮液。 46. 如實施例45之方法，其中該標靶選自肺、肝臟、脾、心臟、腦、淋巴結、膀胱、腎臟及胰臟。 47. 如實施例43至46中任一項之方法，其中該懸浮液係肌內、鼻內、靜脈內、皮下或腫瘤內投與。 48. 一種製備式III化合物，III 或其醫藥學上可接受之鹽之方法，該方法包含使式IV-1化合物與式IV-2化合物在熱存在下接觸：以提供第一中間產物，且其中使該第一中間產物與鉑觸媒及第一酸在氫氣存在下接觸以提供第二中間產物，且其中使該第二中間產物與第二酸接觸以提供該式III化合物，其中 M^1’和M^2’獨立地選自鍵或視情況經取代之C₂-C₁₀脂族基； L^1'及L^2'各自獨立地選自鍵、-OC(O)-及-C(O)O-； T^1'及T^2'各自獨立地選自視情況經取代之C₂-C₂₀脂族基； L^3’為視情況經取代之C₁-C₆脂族基； LG¹為適宜之脫離基；且 PG為適宜之醇保護基。 49. 如實施例48之方法，其中式IV-2化合物藉由以下方式製備：使式V-1a及式V-1b化合物與式V-2化合物在鈀偶合觸媒、銅(I)輔觸媒及胺鹼存在下接觸，以提供該式IV-2化合物，其中LG²為適宜之脫離基。 50. 一種製備式III-1化合物，III-1 或其醫藥學上可接受之鹽之方法，該方法包含使式IV-1化合物與式IV-3化合物在熱存在下接觸：以提供第一中間產物，且其中使該第一中間產物與酸接觸，以提供該式III-1化合物，其中 M^1’和M^2’各自獨立地選自鍵或視情況經取代之C₂-C₁₀脂族基； T^1’及T^2’各自獨立地選自視情況經取代之C₂-C₂₀脂族基； L^3’為視情況經取代之C₁-C₆脂族基； LG¹為適宜之脫離基；且 PG為適宜之醇保護基。EXEMPLARY EMBODIMENTS Although not limiting, the following numbered examples are illustrative of certain aspects of the present disclosure: 1. A compound of formula I, I or a pharmaceutically acceptable salt thereof, wherein^M1 and^M2 are each -(_CH2 )_p1- ;^L1 and^L2 are each selected from a bond, -OC(O)-, and -C(O)O-;^T1 and^T2 are each:^L3 is an optionally substituted_C1 -_C6 aliphatic group;^G1 is -OH or -N(^Ra )₂ , and^Ra is H, an optionally substituted_C1 -_C6 aliphatic group, or an optionally substituted_C3 -_C6 cycloaliphatic group; p1 is an integer selected from 2 to 20, inclusive; p2 and p3 are each an integer selected from 1 to 20, inclusive, and p2 and p3 are the same. 2. The compound of Example 1, wherein p1 is an integer selected from 2 to 10, inclusive. 3. The compound of Example 1 or 2, wherein^M1 is -(_CH2 )_4-6- and^M2 is -(_CH2 )_4-6- . 4. The compound of any one of Examples 1 to 3, wherein^M1 and^M2 are the same. 5. The compound of any one of Examples 1 to 4, wherein^T1 and^T2 are the same. 6. The compound of any one of Examples 1 to 5, wherein^L1 and^L2 are the same. 7. The compound of any one of Examples 1 to 6, wherein p2 and p3 are each an integer selected from 3, 4, 5, 6, 7, 8, or 9. 8. The compound of Example 7, wherein p2 and p3 are each an integer selected from 4, 5, or 6. 9. The compound of Example 1, wherein^T1 and^T2 are each selected from: 、 、 、 、 and 10. The compound of Example 1, wherein M¹ -L¹ -T¹ is represented by: wherein p2 and p3 are each 4, 5 or 6. 11. The compound of embodiment 1 or 10, wherein M² -L² -T² is represented by: wherein p2 and p3 are each 4, 5 or 6. 12. The compound of Example 1, wherein M¹ -L¹ -T¹ and M² -L² -T² are each selected from: 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 and 13. The compound of any one of Examples 1 to 12, wherein -L³ -G¹ is -(CH₂ )₃ -OH or -(CH₂ )₄ -OH. 14. The compound of Example 1, wherein the compound is represented by Formula II-1: II-1 wherein * represents a stereopure carbon atom. 15. The compound of any one of Examples 1 to 14, wherein the compound is characterized by having a Wiener path between about 15,000 and 30,000 and an MMFF94 energy between -15 and 0. 16. The compound of any one of Examples 1 to 15, wherein the compound is characterized by having a k between 0.1 and 0.2. 17. The compound of Example 1, wherein the compound is selected from Table 1. 18. A pharmaceutical composition comprising the compound of any one of Examples 1 to 17 and a nucleic acid. 19. The pharmaceutical composition of Example 18, further comprising one or more of the following: a neutral lipid, a polymer-conjugated lipid, a steroid, or an anionic amphiphile. 20. The pharmaceutical composition of Example 19, further comprising a neutral lipid. 21. The pharmaceutical composition of embodiment 19 or 20, wherein the neutral lipid is a phospholipid. 22. The pharmaceutical composition of embodiment 21, wherein the phospholipid is or comprises distearylphosphatidylcholine (DSPC), dioleoylphosphatidylcholine (DOPC), dimyristoylphosphatidylcholine (DMPC), dipentadecanoylphosphatidylcholine, dilauroylphosphatidylcholine, dipalmitoylphosphatidylcholine (DPPC), diarachidylphosphatidylcholine (DAPC), dibehenylphosphatidylcholine (DBPC), dioctadecanoylphosphatidylcholine (D ... Acylphosphatidylcholine (DTPC), di-tetracosylphosphatidylcholine (DLPC), palmitoyloleyl-phosphatidylcholine (POPC), 1,2-di-O-octadecenyl-sn-glycero-3-phosphocholine (18:0 diether PC), 1-oleyl-2-cholestyryl hemisuccinyl-sn-glycero-3-phosphocholine (OChemsPC), 1-hexadecyl-sn-glycero-3-phosphocholine (C16 Lyso PC), dioleylphosphatidylethanolamine (DOPE), distearylphosphatidylethanolamine (DSPE), dimalmitoylphosphatidylethanolamine (DPPE), dimyristoylphosphatidylethanolamine (DMPE), dilaurylphosphatidylethanolamine (DLPE), diphytylphosphatidylethanolamine (DPyPE), 1,2-bis-(9Z-octadecenyl)-sn-glycero-3-phosphocholine (DOPG), 1,2-dipalmitoyl-sn-glycero-3-phospho-(1′-rac-glycerol) (DPPG), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine (POPE), or N-palmitoyl-D-erythro-sphingamidoylphosphocholine (SM). 23. The pharmaceutical composition of any one of Examples 19 to 22, further comprising a polymer-conjugated lipid. 24. The pharmaceutical composition of Example 23, wherein the polymer-conjugated lipid is a lipid comprising a polymer, and the polymer is polyethylene glycol. 25. The pharmaceutical composition of Example 24, wherein the polymer-coupled lipid is selected from PEG-DAG, PEG-PE, PEG-S-DAG, PEG2000-DMG, PEG-S-DMG, PEG-cer, PEG dialkoxypropylcarbamate (e.g., ω-methoxy(polyethoxy)ethyl-N-(2,3-di-tetradecyloxy)propyl)carbamate or 2,3-di-tetradecyloxypropyl-N-(ω-methoxy(polyethoxy)ethyl)carbamate), and combinations thereof. 26. The pharmaceutical composition of any one of Examples 19 to 25, further comprising a steroid. 27. The pharmaceutical composition of Example 26, wherein the steroid is a sterol. 28. The pharmaceutical composition of Example 27, wherein the sterol is cholesterol. 29. The pharmaceutical composition of any one of Examples 19 to 28, further comprising an anionic amphiphile. 30. The pharmaceutical composition of Example 29, wherein the anionic amphiphile is selected from DMGS, DPGS, and DMGS. 31. The pharmaceutical composition of any one of Examples 19 to 22 and 26 to 29, wherein the pharmaceutical composition comprises a neutral lipid, a steroid, and an anionic amphiphile, and the pharmaceutical composition does not include a polymer-conjugated lipid. 32. The pharmaceutical composition of any one of Examples 18 to 31, wherein the nucleic acid is RNA. 33. The pharmaceutical composition of Example 32, wherein the RNA is modRNA, saRNA, taRNA, or uRNA. 34. The pharmaceutical composition of any one of Examples 18 to 31, wherein the nucleic acid is DNA. 35. The pharmaceutical composition of any one of Examples 18 to 31, wherein the nucleic acid is a mixture of RNA and DNA. 36. The pharmaceutical composition of any one of Examples 19 to 35, wherein the pharmaceutical composition comprises: about 30 mol% to about 60 mol% of the compound; about 20 mol% to about 60 mol% of the steroid; about 1 mol% to about 2 mol% of the polymer-conjugated lipid; and about 5 mol% to about 20 mol% of the neutral lipid. 37. The pharmaceutical composition of any one of Examples 19 to 35, wherein the pharmaceutical composition comprises: about 30 mol% to about 60 mol% of the compound; about 20 mol% to about 60 mol% of the steroid; about 0.5 to about 10 mol% of the anionic amphiphile; and about 5 mol% to about 20 mol% of the neutral lipid. 38. The pharmaceutical composition of Example 37, wherein the pharmaceutical composition does not include a polymer-coupled lipid. 39. A suspension comprising a dispersed phase and an aqueous phase, wherein the dispersed phase comprises the pharmaceutical composition of any one of Examples 18 to 38 in the form of particles. 40. The suspension of Example 39, wherein the particles have an average diameter of about 50 nm to about 200 nm. 41. The suspension of Example 40, wherein the aqueous phase comprises less than 20% of the total concentration of nucleic acids in the suspension. 42. The suspension of any one of Examples 39 to 41, wherein the aqueous phase is substantially free of the nucleic acids. 43. A method of treating a disease, disorder, or condition comprising administering to a subject the suspension of any one of Examples 39 to 42. 44. The method of Example 43, wherein the disease, disorder, or condition is selected from an infectious disease, cancer, a genetic disorder, an autoimmune disease, or a rare disease. 45. A method of increasing or causing an increase in RNA expression in a target in a subject, comprising administering to the subject the suspension of any one of Examples 39 to 42. 46. The method of embodiment 45, wherein the target is selected from the group consisting of lung, liver, spleen, heart, brain, lymph nodes, bladder, kidney, and pancreas. 47. The method of any one of embodiments 43 to 46, wherein the suspension is administered intramuscularly, intranasally, intravenously, subcutaneously, or intratumorally. 48. A method for preparing a compound of formula III, III or a pharmaceutically acceptable salt thereof, which comprises contacting a compound of formula IV-1 with a compound of formula IV-2 in the presence of heat: to provide a first intermediate product, and wherein the first intermediate product is contacted with a platinum catalyst and a first acid in the presence of hydrogen to provide a second intermediate product, and wherein the second intermediate product is contacted with a second acid to provide the compound of formula III, wherein M^1' and M^2' are independently selected from a bond or an optionally substituted C₂ -C₁₀ aliphatic group; L^{1 '} and L^{2 '} are each independently selected from a bond, -OC(O)-, and -C(O)O-; T^{1 '} and T^{2 '} are each independently selected from an optionally substituted C₂ -C₂₀ aliphatic group; L^3' is an optionally substituted C₁ -C₆ aliphatic group; LG¹ is a suitable leaving group; and PG is a suitable alcohol protecting group. 49. The method of Example 48, wherein the compound of formula IV-2 is prepared by reacting the compounds of formula V-1a and formula V-1b with the compound of formula V-2 In the presence of a palladium coupling catalyst, a copper (I) cocatalyst and an amine base, the compound of formula IV-2 is provided, wherein LG² is a suitable deionized group. 50. A method for preparing a compound of formula III-1, III-1 or a pharmaceutically acceptable salt thereof, the method comprising contacting a compound of formula IV-1 with a compound of formula IV-3 in the presence of heat: to provide a first intermediate product, and wherein the first intermediate product is contacted with an acid to provide the compound of formula III-1, wherein M^1' and M^2' are each independently selected from a bond or an optionally substituted C₂ -C₁₀ aliphatic group; T^1' and T^2' are each independently selected from an optionally substituted C₂ -C₂₀ aliphatic group; L^3' is an optionally substituted C₁ -C₆ aliphatic group; LG¹ is a suitable leaving group; and PG is a suitable alcohol protecting group.

實例如下文實例中所述，在某些示範性實施例中，根據以下一般程序製備化合物。應瞭解，儘管一般方法描繪本揭示案之某些化合物之合成，但一般熟習此項技術者已知之以下一般方法及其他方法可適用於所有化合物以及這些化合物中之各者之子類及種類，如本文所述。As described in theExamples below, in certain exemplary embodiments, compounds were prepared according to the following general procedures. It should be understood that although the general methods describe the synthesis of certain compounds of the present disclosure, the following general methods and other methods known to those of ordinary skill in the art can be applied to all compounds, as well as subclasses and species of each of these compounds, as described herein.

縮寫列表aq. =水溶液 DMGS =二肉荳蔻醯基甘油半琥珀酸酯 DPGS =二棕櫚醯基甘油半琥珀酸酯 DSGS =二硬脂醯基甘油基半琥珀酸酯 DSPC =二硬脂醯基磷脂醯膽鹼 equiv. =當量 h或hr =小時(hour/hours) HRMS =高解析度質譜 LNP =脂質奈米粒子 NMR =核磁共振 Otf =三氟甲磺酸酯 PEG =聚乙二醇 rt =室溫 sat. =飽和 TBS =第三丁基二甲基矽基醚 THF =四氫呋喃 TLC =薄層層析Abbreviations: aq. = aqueous solution; DMGS = dimyristylglycerol hemisuccinate; DPGS = disaldicarboxylic acid glycerol hemisuccinate; DSGS = distearylglycerol hemisuccinate; DSPC = distearylphosphatidylcholine; equiv. = equivalents; h or hr = hour/hour; HRMS = high-resolution mass spectrometry; LNP = lipid nanoparticle; NMR = nuclear magnetic resonance; Otf = trifluoromethanesulfonate; PEG = polyethylene glycol; rt = room temperature; sat. = saturated; TBS = tert-butyldimethylsilyl ether; THF = tetrahydrofuran; TLC = thin layer chromatography.

合成脂質之一般合成途徑：用於合成本文所述化合物之一般合成途徑包含兩個線性步驟，Sonogashira偶合，隨後烷基化、還原及TBS去保護。下文提供例示實例合成方法之方案。其中A為適宜之脫離基(例如鹵素，諸如I、Br、Cl或三氟甲磺酸酯，亦即-O-S(O)-CF₃)，且X、Y及Z中之各者在各情況下獨立地為鍵或-(CH₂)_1-4-。General Synthetic Route for Synthetic Lipids: The general synthetic route for the compounds described herein comprises two linear steps, a Sonogashira coupling, followed by alkylation, reduction, and TBS deprotection. Schemes illustrating exemplary synthetic methods are provided below. wherein A is a suitable ionizing group (e.g., a halogen such as I, Br, Cl, or triflate, i.e., —OS(O)—CF₃ ), and each of X, Y, and Z is independently a bond or —(CH₂ )_1-4 —.

在一些實施例中，本文所述之化合物亦可根據以下製程來製備：其中A為適宜之脫離基(例如鹵素，諸如I、Br、Cl或三氟甲磺酸酯，亦即-O-S(O)-CF₃)，且X、Y及Z中之各實例在各情況下獨立地為鍵或-(CH₂)_1-4-。In some embodiments, the compounds described herein can also be prepared according to the following process: wherein A is a suitable ionizing group (e.g., a halogen such as I, Br, Cl, or triflate, i.e., -OS(O)-CF₃ ), and each instance of X, Y, and Z is independently a bond or -(CH₂ )_1-4 -.

實例1：吡啶-3,5-二基雙(丁-3-炔-4,1-二基)雙(2-庚基壬酸酯)：向帶有磁力攪拌棒之25 mL火焰乾燥之Schlenk燒瓶中裝入2,5-二溴吡啶(473.0 mg，2.0 mmol，1.0 equiv.)、碘化銅(I) (38.0 mg，0.2 mmol，0.1 equiv.)及四(三苯基膦)-鈀(116.0 mg，0.1 mmol，0.05 equiv.)。接著將燒瓶在高真空下小心地抽真空且置於氬氣下。將三乙胺(10.0 mL)添加至固體中，且接著逐滴添加2-庚基壬酸丁-3-炔-1-基酯(1.8 g，6 mmol，3.0 equiv.)。將反應混合物在60℃下加熱18 h。藉由TLC監測反應之進展。在減壓下濃縮反應混合物且純化粗混合物，且藉由管柱層析在矽膠上使用己烷/乙酸乙酯(95:5，v/v)作為溶析液純化粗混合物。獲得呈棕色液體狀之產物(0.83 g，60%產率)。¹H NMR (501 MHz, CDCl₃) δ 8.49 (d,J= 2.0 Hz, 2H), 7.64 (t,J= 2.0 Hz, 1H), 4.26 (t,J= 6.7 Hz, 4H), 2.76 (t,J= 6.7 Hz, 4H), 2.35 (tt,J= 8.9, 5.3 Hz, 2H), 1.66–1.55 (m, 4H), 1.45 (ddt,J= 14.7, 11.4, 5.4 Hz, 4H), 1.33–1.12 (m, 40H), 0.86 (t,J= 6.9 Hz, 12H).¹³C NMR (126 MHz, CDCl₃) δ 176.48, 150.92, 140.91, 120.19, 90.08, 78.14, 61.73, 45.89, 32.63, 31.94, 29.67, 29.27, 27.61, 27.12, 22.76, 20.21, 14.22。Example 1: Pyridine-3,5-diylbis(but-3-yn-4,1-diyl)bis(2-heptylnonanoate): A 25 mL flame-dried Schlenk flask equipped with a magnetic stir bar was charged with 2,5-dibromopyridine (473.0 mg, 2.0 mmol, 1.0 equiv.), copper(I) iodide (38.0 mg, 0.2 mmol, 0.1 equiv.), and tetrakis(triphenylphosphine)-palladium (116.0 mg, 0.1 mmol, 0.05 equiv.). The flask was then carefully evacuated under high vacuum and placed under an atmosphere of nitrogen. Triethylamine (10.0 mL) was added to the solid, followed by the dropwise addition of but-3-yn-1-yl 2-heptylnonanoate (1.8 g, 6 mmol, 3.0 equiv.). The reaction mixture was heated at 60°C for 18 h. The progress of the reaction was monitored by TLC. The reaction mixture was concentrated under reduced pressure, and the crude mixture was purified by column chromatography on silica gel using hexane/ethyl acetate (95:5, v/v) as the eluent. The product was obtained as a brown liquid (0.83 g, 60% yield).¹ H NMR (501 MHz, CDCl₃ ) δ 8.49 (d,J = 2.0 Hz, 2H), 7.64 (t,J = 2.0 Hz, 1H), 4.26 (t,J = 6.7 Hz, 4H), 2.76 (t,J = 6.7 Hz, 4H), 2.35 (tt,J13C^​ NMR (126 MHz, CDCl₃ ) δ 176.48, 150.92, 140.91, 120.19, 90.08, 78.14, 61.73, 45.89, 32.63, 31.94, 29.67, 29.27, 27.61, 27.12, 22.76, 20.21, 14.22.

實例2：吡啶-3,5-二基雙(己-5-炔-6,1-二基)雙(2-己基癸酸酯)：本實例之化合物係以與實例1中所述之方法類似之方法，改為使用2-己基癸酸己-5-炔-1-基酯來製備，以提供呈棕色液體狀之標題化合物(1.1 g，92%產率)。¹H NMR (501 MHz, CDCl₃) δ 8.43 (d,J= 2.0 Hz, 2H), 7.59 (t,J= 2.1 Hz, 1H), 4.09 (t,J= 6.4 Hz, 4H), 2.42 (t,J= 7.0 Hz, 4H), 2.28 (tt,J= 9.1, 5.3 Hz, 2H), 1.76 (dq,J= 8.9, 6.5 Hz, 4H), 1.69–1.60 (m, 4H), 1.60–1.49 (m, 4H), 1.39 (dp,J= 14.3, 5.0 Hz, 4H), 1.22 (dd,J= 13.1, 6.1 Hz, 40H), 0.82 (td,J= 6.9, 2.3 Hz, 12H)。¹³C NMR (126 MHz, CDCl₃) δ 176.56, 150.49, 140.69, 120.44, 93.57, 77.32, 63.39, 45.84, 32.58, 31.89, 31.74, 29.60, 29.49, 29.29, 29.26, 28.01, 27.53, 27.49, 25.13, 22.70, 22.63, 19.13, 14.13, 14.08。HRMS m/z (ESI)：C₄₉H₈₂O₄N計算值[M+H]：748.62384；實測值：748.62334。Example 2: Pyridine-3,5-diylbis(hex-5-yn-6,1-diyl)bis(2-hexyldecanoate): The compound of this example was prepared by a method similar to that described in Example 1, except that hex-5-yn-1-yl 2-hexyldecanoate was used to provide the title compound (1.1 g, 92% yield) as a brown liquid.¹ H NMR (501 MHz, CDCl₃ ) δ 8.43 (d,J = 2.0 Hz, 2H), 7.59 (t,J = 2.1 Hz, 1H), 4.09 (t,J = 6.4 Hz, 4H), 2.42 (t,J = 7.0 Hz, 4H), 2.28 (tt,J1.22​ (dd,J = 13.1, 6.1 Hz, 40H), 0.82 (td,J = 6.9, 2.3 Hz, 12H).¹³ C NMR (126 MHz, CDCl₃ ) δ 176.56, 150.49, 140.69, 120.44, 93.57, 77.32, 63.39, 45.84, 32.58, 31.89, 31.74, 29.60, 29.49, 29.29, 29.26, 28.01, 27.53, 27.49, 25.13, 22.70, 22.63, 19.13, 14.13, 14.08. HRMS m/z (ESI): calcd. for C₄₉ H₈₂ O₄ N [M+H]: 748.62384; found: 748.62334.

實例3：吡啶-3,5-二基雙(己-5-炔-6,1-二基)雙(2-庚基壬酸酯)：本實例之化合物係以與實例1中所述之方法類似之方法，改為使用2-庚基壬酸己-5-炔-1-基酯來製備，以提供呈棕色液體狀之標題化合物(1.4 g，94%產率)。Example 3: Pyridine-3,5-diylbis(hex-5-yn-6,1-diyl)bis(2-heptylnonanoate): The compound in this example was prepared by a method similar to that described in Example 1, except that hex-5-yn-1-yl 2-heptylnonanoate was used to provide the title compound (1.4 g, 94% yield) as a brown liquid.

實例4：(1-(4-羥基丁基)哌啶-3,5-二基)雙(己烷-6,1-二基)雙(2-己基癸酸酯)：PIL4677Example 4: (1-(4-Hydroxybutyl)piperidin-3,5-diyl)bis(hexane-6,1-diyl)bis(2-hexyldecanoate): PIL4677

向帶有磁性攪拌棒之25 mL火焰乾燥圓底燒瓶中裝入吡啶-3,5-二基雙(己-5-炔-6,1-二基)雙(2-己基癸酸酯) (1.0 g，1.45 mmol，1.0 equiv.)及1-溴-4-(第三丁基二甲基矽氧基)-丁烷(1.1 g，3.9 mmol，2.7 equiv.)。接著將反應混合物在110℃下加熱16 h。藉由TLC監測反應之進展。A 25 mL flame-dried round-bottom flask equipped with a magnetic stir bar was charged with pyridine-3,5-diylbis(hex-5-yn-6,1-diyl)bis(2-hexyldecanoate) (1.0 g, 1.45 mmol, 1.0 equiv.) and 1-bromo-4-(tert-butyldimethylsilyloxy)-butane (1.1 g, 3.9 mmol, 2.7 equiv.). The reaction mixture was then heated at 110°C for 16 h. The reaction progress was monitored by TLC.

16 h後，將反應混合物在rt下冷卻且以定量產率獲得吡啶鎓鹽。向此粗製吡啶鎓鹽中添加乙酸(1.5 mL)及PtO₂(22.5 mg，0.1 mmol，0.1 equiv.)。將反應混合物在40℃下於H₂氣氛(1 atm)下加熱18 h。藉由TLC監測反應之進展。After 16 h, the reaction mixture was cooled at room temperature and the pyridinium salt was obtained in quantitative yield. To this crude pyridinium salt was added acetic acid (1.5 mL) and_PtO₂ (22.5 mg, 0.1 mmol, 0.1 equiv.). The reaction mixture was heated at 40°C under a_H₂ atmosphere (1 atm) for 18 h. The progress of the reaction was monitored by TLC.

18 h後，將反應混合物在rt下冷卻。向此反應混合物中添加THF (5 mL)及5 mL HCl水溶液(10%)。將混合物在rt下攪拌30 min。接著將反應混合物用Et₂O (25 mL)稀釋且收集有機相。用Et₂O (3×20 mL)萃取水相。合併之有機相經Na₂SO₄乾燥，過濾且在減壓下濃縮。在矽膠管柱層析上使用乙酸乙酯/EtOH (95:5 -＞ 85:15，v/v)作為溶析液純化粗混合物。產物以HCl鹽形式分離。將HCl鹽用飽和NaHCO₃水溶液處理且獲得呈淺黃色油狀之脂質(0.36 g，44%產率，經三個步驟)。¹H NMR (501 MHz, CDCl₃) δ 4.02 (t,J= 6.6 Hz, 4H), 3.43 (q,J= 7.0 Hz, 1H), 3.03–2.96 (m, 2H), 2.80 (dd,J= 12.1, 3.7 Hz, 1H), 2.45 (dd,J= 12.2, 6.4 Hz, 1H), 2.26 (tt,J= 9.0, 5.3 Hz, 2H), 2.21–2.09 (m, 1H), 2.06 (t,J= 11.5 Hz, 2H), 1.80 (d,J= 12.8 Hz, 1H), 1.55 (dp,J= 16.0, 7.2 Hz, 8H), 1.45–1.00 (m, 65H), 0.93–0.71 (m, 12H), 0.55 (q,J= 12.1 Hz, 1H)。¹³C NMR (126 MHz, CDCl₃) δ 176.68, 65.89, 64.11, 60.39, 53.06, 51.98, 45.91, 38.96, 37.44, 36.15, 34.76, 32.96, 32.87, 32.63, 31.94, 31.78, 29.63, 29.60, 29.52, 29.45, 29.32, 29.30, 28.81, 28.79, 27.54, 27.50, 27.29, 26.71, 26.07, 26.02, 22.73, 22.66, 15.33, 14.26, 14.16, 14.13。HRMS m/z (ESI)：C₅₃H₁₀₄NO₅計算值[M+H]：834.79090；實測值：834.79011。After 18 h, the reaction mixture was cooled at rt. THF (5 mL) and 5 mL of aqueous HCl (10%) were added to the reaction mixture. The mixture was stirred at rt for 30 min. The reaction mixture was then diluted with_Et2O (25 mL) and the organic phase was collected. The aqueous phase was extracted with_Et2O (₃ × 20 mL). The combined organic phases were dried over_Na2SO4 , filtered, and concentrated under reduced pressure. The crude mixture was purified by silica gel column chromatography using ethyl acetate/EtOH (95:5 -> 85:15, v/v) as the eluent. The product was isolated as the HCl salt. The HCl salt was treated with saturated aqueous NaHCO₃ solution and the lipid was obtained as a light yellow oil (0.36 g, 44% yield over three steps).¹ H NMR (501 MHz, CDCl₃ ) δ 4.02 (t,J = 6.6 Hz, 4H), 3.43 (q,J = 7.0 Hz, 1H), 3.03–2.96 (m, 2H), 2.80 (dd,J = 12.1, 3.7 Hz, 1H), 2.45 (dd,1.55​​​ (dp,J = 16.0, 7.2 Hz, 8H), 1.45–1.00 (m, 65H), 0.93–0.71 (m, 12H), 0.55 (q,J = 12.1 Hz, 1H).¹³ C NMR (126 MHz, CDCl₃ ) δ 176.68, 65.89, 64.11, 60.39, 53.06, 51.98, 45.91, 38.96, 37.44, 36.15, 34.76, 32.96, 32.87, 32.63, 31.94, 31.78, 29.63, 29.60, 29.52, 29.45, 29.32, 29.30, 28.81, 28.79, 27.54, 27.50, 27.29, 26.71, 26.07, 26.02, 22.73, 22.66, 15.33, 14.26, 14.16, 14.13. HRMS m/z (ESI): Calcd. for C₅₃ H₁₀₄ NO₅ [M+H]: 834.79090; Found: 834.79011.

實例5：(1-(3-羥基丙基)哌啶-3,5-二基)雙(己烷-6,1-二基)雙(2-庚基壬酸酯)：PIL 3677Example 5: (1-(3-Hydroxypropyl)piperidin-3,5-diyl)bis(hexane-6,1-diyl)bis(2-heptylnonanoate): PIL 3677

本實例之化合物係根據實例4中所述之方法製備，其中使吡啶-3,5-二基雙(己-5-炔-6,1-二基)雙(2-庚基壬酸酯)與1-溴-3-(第三丁基二甲基矽氧基)-丙烷反應，以提供呈淺黃色油狀之標題化合物(0.17 g，48%產率，經三個步驟)。¹H NMR (501 MHz, CDCl₃) δ 4.06 (t,J= 6.6 Hz, 4H), 3.82–3.75 (m, 2H), 3.02 (d,J= 10.1 Hz, 2H), 2.58 (t,J= 5.5 Hz, 2H), 2.31–2.30 (m, 2H), 1.85–1.77 (m, 1H), 1.72 (p,J= 5.3 Hz, 2H), 1.66–1.01 (m, 73H), 0.87 (t,J= 6.9 Hz, 12H), 0.47 (q,J= 11.8 Hz, 1H)。¹³C NMR (126 MHz, CDCl₃) δ 176.86, 64.24, 60.72, 60.53, 59.43, 46.01, 38.11, 36.24, 34.73, 32.70, 31.96, 29.75, 29.67, 29.31, 28.85, 27.62, 27.23, 26.87, 26.09, 22.79, 21.19, 14.35, 14.24。The compound of this example was prepared according to the method described in Example 4, wherein pyridine-3,5-diylbis(hex-5-yn-6,1-diyl)bis(2-heptylnonanoate) was reacted with 1-bromo-3-(tert-butyldimethylsilyloxy)-propane to provide the title compound as a light yellow oil (0.17 g, 48% yield over three steps).¹ H NMR (501 MHz, CDCl₃ ) δ 4.06 (t,J = 6.6 Hz, 4H), 3.82–3.75 (m, 2H), 3.02 (d,J = 10.1 Hz, 2H), 2.58 (t,J = 5.5 Hz, 2H), 2.31–2.30 (m, 2H), 1.85–1.77 (m, 1H), 1.72 (p,J = 5.3 Hz, 2H), 1.66–1.01 (m, 73H), 0.87 (t,J = 6.9 Hz, 12H), 0.47 (q,J = 11.8 Hz, 1H).¹³ C NMR (126 MHz, CDCl₃ ) δ 176.86, 64.24, 60.72, 60.53, 59.43, 46.01, 38.11, 36.24, 34.73, 32.70, 31.96, 29.75, 29.67, 29.31, 28.85, 27.62, 27.23, 26.87, 26.09, 22.79, 21.19, 14.35, 14.24.

實例6：(1-(3-羥基丙基)哌啶-3,5-二基)雙(丁烷-4,1-二基)雙(2-庚基壬酸酯)：CPIL 3477本實例之化合物係根據實例4中所述之方法製備，其中使吡啶-3,5-二基雙(丁-3-炔-4,1-二基)雙(2-庚基壬酸酯)與1-溴-3-(第三丁基二甲基矽氧基)-丙烷，以提供呈淺黃色油狀之標題化合物(0.15 g，43%產率，經三個步驟)。¹H NMR (501 MHz, CDCl₃) δ 4.06 (t,J= 6.6 Hz, 4H), 3.79 (dt,J= 11.8, 5.4 Hz, 2H), 3.67–3.45 (m, 1H), 3.02 (d,J= 10.3 Hz, 2H), 2.58 (s, 2H), 2.31 (tt,J= 8.9, 5.3 Hz, 2H), 1.94–1.76 (m, 1H), 1.71 (s, 2H), 1.59 (dt,J= 13.2, 6.5 Hz, 10H), 1.50–1.07 (m, 54H), 0.87 (t,J= 6.9 Hz, 12H), 0.49 (q,J= 11.9 Hz, 1H)。¹³C NMR (126 MHz, CDCl₃) δ 176.83, 64.04, 45.99, 34.37, 32.67, 31.97, 29.68, 29.32, 29.17, 27.63, 23.34, 22.79, 14.25。Example 6: (1-(3-Hydroxypropyl)piperidin-3,5-diyl)bis(butane-4,1-diyl)bis(2-heptylnonanoate): CPIL 3477 The compound of this example was prepared according to the method described in Example 4, wherein pyridine-3,5-diylbis(but-3-yn-4,1-diyl)bis(2-heptylnonanoate) was reacted with 1-bromo-3-(tert-butyldimethylsilyloxy)-propane to provide the title compound as a light yellow oil (0.15 g, 43% yield over three steps).¹ H NMR (501 MHz, CDCl₃ ) δ 4.06 (t,J = 6.6 Hz, 4H), 3.79 (dt,J = 11.8, 5.4 Hz, 2H), 3.67–3.45 (m, 1H), 3.02 (d,J = 10.3 Hz, 2H), 2.58 (s, 2H), 2.31 (tt,J = 8.9, 5.3 Hz, 2H), 1.94–1.76 (m, 1H), 1.71 (s, 2H), 1.59 (dt,J = 13.2, 6.5 Hz, 10H), 1.50–1.07 (m, 54H), 0.87 (t,J = 6.9 Hz, 12H), 0.49 (q,J = 11.9 Hz, 1H).¹³ C NMR (126 MHz, CDCl₃ ) δ 176.83, 64.04, 45.99, 34.37, 32.67, 31.97, 29.68, 29.32, 29.17, 27.63, 23.34, 22.79, 14.25.

實例7：(1-(4-羥基丁基)哌啶-3,5-二基)雙(己烷-6,1-二基)雙(2-辛基癸酸酯) PIL4668Example 7: (1-(4-Hydroxybutyl)piperidin-3,5-diyl)bis(hexane-6,1-diyl)bis(2-octyldecanoate) PIL4668

實例8：環狀可離子化脂質之生物活性本實例描述自如本文所述之哌啶可離子化脂質、膽固醇、DSPC及聚合物偶聯脂質製備脂質奈米粒子(LNP)組合物。Example 8: Biological Activity of Cyclic Ionizable Lipids This example describes the preparation of lipid nanoparticle (LNP) compositions from piperidine ionizable lipids as described herein, cholesterol, DSPC, and polymer-conjugated lipids.

研究設計本實例報告一系列經測試之組合物，其中哌啶可離子化脂質介於29.9 mol%與66.2 mol%之間，膽固醇介於17.7 mol%與57.1 mol%之間，DSPC介於2.6 mol%與23.8 mol%之間且PEG-脂質為1.8 mol%。Study Design This example reports on a series of compositions tested, in which the piperidine ionizable lipid ranged from 29.9 mol% to 66.2 mol%, cholesterol ranged from 17.7 mol% to 57.1 mol%, DSPC ranged from 2.6 mol% to 23.8 mol%, and PEG-lipid was 1.8 mol%.

LNP之產生製備具有以下濃度之儲備溶液濃度：mRNA：0.22 mg/mL於去離子水中，哌啶可離子化脂質：50 mM (經75 mM乙酸酸化)，膽固醇：50 mM，DSPC：33.33 mM，PEG-脂質：8 mM，其中乙醇用作所有脂質之溶劑。滴加混合製程：將哌啶可離子化脂質添加至孔板中，隨後依次添加膽固醇、DSPC及PEG-脂質(用於參考調配物)。添加適量乙醇至33 μL乙醇之最終體積。添加mRNA溶液至0.055 mg/mL之最終濃度，隨後添加緩衝液(35 mM Tris:Hac，pH = 5.5)。mRNA及緩衝溶液均完成200 μL之最終體積。取出等分試樣且使用緩衝液(35 mM Tris:Hac，pH = 7.5)進一步逐步稀釋。此時使用Wyatt Dynapro III動態光散射儀器監測粒徑。LNPproduction : A stock solution with the following concentrations was prepared: mRNA: 0.22 mg/mL in deionized water, piperidine-ionizable lipid: 50 mM (acidified with 75 mM acetic acid), cholesterol: 50 mM, DSPC: 33.33 mM, and PEG-lipid: 8 mM. Ethanol was used as a solvent for all lipids. A drop-by-drop mixing process was used: piperidine-ionizable lipid was added to the well plate, followed by cholesterol, DSPC, and PEG-lipid (for a reference formulation). Ethanol was added to a final volume of 33 μL. The mRNA solution was added to a final concentration of 0.055 mg/mL, followed by the addition of buffer (35 mM Tris:Hac, pH 5.5). Both mRNA and buffer were added to a final volume of 200 μL. Aliquots were removed and further diluted with buffer (35 mM Tris:Hac, pH 7.5). Particle size was monitored using a Wyatt Dynapro III dynamic light scattering instrument.

在進一步分析中取消選擇具有低於40nm之極小尺寸或高於250nm之大尺寸之材料。Materials with extremely small dimensions below 40 nm or large dimensions above 250 nm were eliminated from further analysis.

對於活體外表現，將10 μL樣品轉移至新孔板中，且添加10 μL人類血清。將混合物在室溫下培育30分鐘，接著轉染。將第二個10 μL樣品轉移至另一孔板中，且添加10 μL水。接著，對於未經血清及含血清之奈米粒子，用每孔50 ng之總mRNA含量轉染細胞。細胞培養物中之最終乙醇體積為0.125%。Forin vitro characterization, 10 μL of sample was transferred to a new plate and 10 μL of human serum was added. The mixture was incubated at room temperature for 30 minutes before transfection. A second 10 μL sample was transferred to another plate and 10 μL of water was added. Next, for both serum-free and serum-containing nanoparticles, cells were transfected with 50 ng of total mRNA per well. The final ethanol content in the cell culture was 0.125%.

在HEK-293及HEPG2細胞中量測活體外表現。In vitro expressionwas measured in HEK-293 and HEPG2 cells.

對表現資料進行分群(n=5)，且根據脂質選擇具有最高表現之群集。下表顯示平均表現水準。The performance data were clustered (n=5), and the cluster with the highest performance was selected based on lipid profile. The table below shows the average performance levels.

脂質LipidsHEK 293HEK 293HepG2HepG2-血清-Serum+血清+Serum-血清-Serum+血清+Serum基準benchmark7.32E+067.32E+061.15E+071.15E+073.90E+063.90E+064.52E+064.52E+06PIL4677PIL4677PIL3677PIL3677PIL4668PIL4668

資料表明，對於包含本文所述之哌啶可離子化脂質、膽固醇、DSPC及PEG-脂質之材料，可獲得粒徑＜=250nm之材料。The data indicate that for materials comprising the piperidine ionizable lipids described herein, cholesterol, DSPC, and PEG-lipids, materials with particle sizes <= 250 nm can be obtained.

實例9：自哌啶可離子化脂質製成之LNP之組合物對於具有最高活性之哌啶可離子化脂質，最佳化脂質組合物。測試了組合物範圍，其中哌啶可離子化脂質介於30 mol%與66 mol%之間，膽固醇介於18 mol%與570 mol%之間，DSPC介於2.6 mol%與24 mol%之間且PEG-脂質為1.8 mol%。Example 9: Composition of LNPs Made from Piperidinium-Ionizable Lipids . The lipid composition was optimized for the piperidinium-ionizable lipid with the highest activity. A range of compositions was tested, including piperidinium-ionizable lipid concentrations between 30 and 66 mol%, cholesterol between 18 and 570 mol%, DSPC between 2.6 and 24 mol%, and PEG-lipid at 1.8 mol%.

如實例8中所概述來製備及分析粒子。Particles were prepared and analyzed as outlined in Example 8.

圖1顯示當在人類血清中預培育脂質奈米粒子之情況下在HepG2細胞中進行測試時，自完全組合基質中之哌啶可離子化脂質、膽固醇、DSPC及PEG-脂質製成之LNP的活體外表現。Figure 1 shows the in vitro performance of LNPs made from piperidine-ionizable lipids, cholesterol, DSPC, and PEG-lipid in a complete combination matrix when tested in HepG2 cells with the lipid nanoparticles preincubated in human serum.

如上文所述製備LNP且監測粒度。大於250nm之LNP用「×」表示且排除在進一步分析之外。將信號正規化為參考=1且分類為淺灰色之之「低活性」，其表現小於參考之10^-0.3、灰色之「良好活性」，其表現介於參考之10^-0.3與10^-0之間，以及深灰色之「高活性」，其表現高於參考。LNPs were prepared as described above and particle size was monitored. LNPs larger than 250 nm were indicated by an "×" and excluded from further analysis. Signals were normalized to a reference of 1 and categorized as "low activity" (light gray), representing less than^10-0.3 of the reference; "good activity" (gray), representing between^10-0.3 and^10-0 of the reference; and "high activity" (dark gray), representing greater than the reference.

資料表明，對於包含哌啶可離子化脂質、膽固醇、DSPC及PEG-脂質之材料，可獲得粒徑＜=250nm之材料。對於PIL3677，對於脂質組合物之幾乎整個範圍，例如具有30 mol%與65 mol%之間的可離子化脂質以及20 mol%與58 mol%之間的膽固醇以及2.5 mol%與24 mol%之間的DSPC，觀測到脂質奈米粒子之膠體性質及高活性。對於PIL4677，具有30 mol%與50 mol%之間的可離子化脂質及35 mol%至50 mol%膽固醇之組合物係較佳的。參見圖1。The data demonstrate that for materials containing piperidine-ionizable lipids, cholesterol, DSPC, and PEG-lipids, particle sizes of ≤250 nm can be obtained. For PIL3677, colloidal properties and high activity of the lipid nanoparticles were observed across nearly the entire range of lipid compositions, for example, with ionizable lipids between 30 and 65 mol%, cholesterol between 20 and 58 mol%, and DSPC between 2.5 and 24 mol%. For PIL4677, compositions with ionizable lipids between 30 and 50 mol% and cholesterol between 35 and 50 mol% were preferred.See Figure 1.

實例10：包含缺乏隱形脂質之哌啶可離子化脂質之LNP在此實例中，脂質奈米粒子(LNP)組合物係自哌啶可離子化脂質(例如，本文所述之化合物)、膽固醇、DSPC及陰離子兩親物產生。Example 10: LNPs Comprising Piperidinium-Ionizable Lipids Lacking Stealth Lipids In this example, lipid nanoparticle (LNP) compositions were generated from piperidine-ionizable lipids (e.g., compounds described herein), cholesterol, DSPC, and anionic amphiphiles.

研究設計由於哌啶可離子化脂質係新穎的且不具有隱形脂質之LNP的存在係未知的，因此先驗脂質組合物可能不會導致脂質奈米粒子之形成，或所形成之彼等脂質奈米粒子可能不具有高活性。因此，測試了一系列組合物，其中哌啶可離子化脂質PIL3677介於25 mol%與55 mol%之間，膽固醇介於17.5 mol%與59.0 mol%之間，DSPC介於2.5 mol%與25.0 mol%之間，且DMGS、DPGS或SMGS用作0.5 mol%與2.5 mol%之間的陰離子兩親物。Study Design: Because piperidine-ionizable lipids are novel and the existence of LNPs without stealth lipids is unknown,a priori lipid compositions may not result in the formation of lipid nanoparticles, or those that do form may not be highly active. Therefore, a series of compositions were tested, with the piperidine-ionizable lipid PIL3677 ranging from 25 mol% to 55 mol%, cholesterol ranging from 17.5 mol% to 59.0 mol%, DSPC ranging from 2.5 mol% to 25.0 mol%, and DMGS, DPGS, or SMGS used as the anionic amphiphile ranging from 0.5 mol% to 2.5 mol%.

如實例8中所述來製備及分析粒子。圖2顯示自哌啶可離子化脂質、膽固醇、DSPC及DMGS (二肉荳蔻醯基甘油半琥珀酸酯)、DPGS (二棕櫚醯基甘油半琥珀酸酯)或SMGS (硬脂醯基肉荳蔻醯基甘油半琥珀酸酯)中之一者作為陰離子兩親物在完全組合基質中製成之LNP的活體外表現。Particles were prepared and analyzed as described in Example 8. Figure 2 shows the in vitro performance of LNPs made from piperidine-ionizable lipids, cholesterol, DSPC, and one of DMGS (dimyristylglycerol hemisuccinate), DPGS (dipalmitoylglycerol hemisuccinate), or SMGS (stearylmyristylglycerol hemisuccinate) as the anionic amphiphile in a complete combination matrix.

如上文所述製備LNP且監測粒度。大於200nm之LNP用「×」表示且排除在進一步分析之外。在未預先與血清在HEK293細胞上培育之情況下獲得活體外表現水準。將信號正規化為參考=1且分類為淺灰色之之「低活性」，其表現小於參考之10^-0.3、灰色之「良好活性」，其表現介於參考之10^-0.3與10⁰之間，以及深灰色之「高活性」，其表現高於參考。LNPs were prepared and particle size monitored as described above. LNPs larger than 200 nm are indicated by an "X" and excluded from further analysis. In vitro performance levels were obtained on HEK293 cells without prior incubation with serum. Signals were normalized to a reference of 1 and categorized as "low activity" (light gray), representing performance less than^10-0.3 of the reference; "good activity" (gray), representing performance between^10-0.3 and¹⁰⁰ of the reference; and "high activity" (dark gray), representing performance greater than the reference.

資料表明，對於包含哌啶可離子化脂質、膽固醇、DSPC及DMGS、DPGS及DSGS中之一者作為陰離子兩親物之材料，可獲得粒徑＜=250nm之材料。當選擇PIL3677時，包含陰離子兩親物之脂質奈米粒子通常展示出與基準相當之活性。較佳組合物具有30 mol%與60 mol%之間的膽固醇。Data indicate that materials containing a piperidine-ionizable lipid, cholesterol, DSPC, and one of DMGS, DPGS, and DSGS as anionic amphiphiles can yield materials with particle sizes ≤ 250 nm. When PIL3677 was selected as the anionic amphiphile, lipid nanoparticles containing the anionic amphiphile generally exhibited comparable activity to the baseline. Optimal compositions had between 30 mol% and 60 mol% cholesterol.

上文所述揭示內容之實施例意欲僅具有示範性，熟習此項技術者將明瞭多種變化及修改。所有此類變化及修改意欲在如任何所附申請專利範圍中所定義之本發明之範圍內。The embodiments disclosed above are intended to be exemplary only, and those skilled in the art will readily appreciate that various variations and modifications may be made thereto. All such variations and modifications are intended to be within the scope of the present invention as defined in any of the appended patent claims.

圖1例示使用本文所述之陽離子或可離子化脂質化合物製備之LNP在HepG2細胞中之活體外表現。 圖2例示使用所述陽離子或可離子化脂質化合物製備之LNP之活體外表現，其中該等LNP包含選自DMGS、DPGS及SMGS之陰離子兩親物。Figure 1 illustrates the in vitro performance of LNPs prepared using the cationic or ionizable lipid compounds described herein in HepG2 cells.Figure 2 illustrates the in vivo performance of LNPs prepared using the cationic or ionizable lipid compounds described herein, wherein the LNPs comprise an anionic amphiphile selected from DMGS, DPGS, and SMGS.

Claims (50)

Translated fromChinese

一種式I化合物，I 或其醫藥學上可接受之鹽，其中 M¹及M²各自為-(CH₂)_p1-； L¹及L²各自選自鍵、-OC(O)-及-C(O)O-； T¹及T²各自為：； L³為視情況經取代之C₁-C₆脂族基； G¹為-OH或-N(R^a)₂，且 R^a為H、視情況經取代之C₁-C₆脂族基或視情況經取代之C₃-C₆環脂族基； p1為選自2-20之整數，包括端值； p2及p3各自為選自1-20之整數，包括端值，且p2及p3係相同的。A compound of formula I, I or a pharmaceutically acceptable salt thereof, wherein^M1 and^M2 are each -(_CH2 )_p1- ;^L1 and^L2 are each selected from a bond, -OC(O)-, and -C(O)O-;^T1 and^T2 are each:^L3 is an optionally substituted_C1 -_C6 aliphatic group;^G1 is -OH or -N(^Ra )₂ , and^Ra is H, an optionally substituted_C1 -_C6 aliphatic group, or an optionally substituted_C3 -_C6 cycloaliphatic group; p1 is an integer selected from 2 to 20, inclusive; p2 and p3 are each an integer selected from 1 to 20, inclusive, and p2 and p3 are the same.如請求項1之化合物，其中p1為選自2-10之整數，包括端值。The compound of claim 1, wherein p1 is an integer selected from 2 to 10, inclusive.如請求項1或2之化合物，其中M¹為-(CH₂)_4-6-且M²為-(CH₂)_4-6-。The compound of claim 1 or 2, wherein M¹ is -(CH₂ )_4-6 - and M² is -(CH₂ )_4-6 -.如請求項1至3中任一項之化合物，其中M¹及M²係相同的。The compound of any one of claims 1 to 3, wherein M¹ and M² are the same.如請求項1至4中任一項之化合物，其中T¹及T²係相同的。The compound of any one of claims 1 to 4, wherein^T1 and^T2 are the same.如請求項1至5中任一項之化合物，其中L¹及L²係相同的。The compound of any one of claims 1 to 5, wherein L¹ and L² are the same.如請求項1至6中任一項之化合物，其中p2及p3各自為選自3、4、5、6、7、8或9之整數。The compound of any one of claims 1 to 6, wherein p2 and p3 are each an integer selected from 3, 4, 5, 6, 7, 8 or 9.如請求項7之化合物，其中p2及p3各自為選自4、5或6之整數。The compound of claim 7, wherein p2 and p3 are each an integer selected from 4, 5 or 6.如請求項1之化合物，其中T¹及T²各自選自：、、、、及。The compound of claim 1, wherein^T1 and^T2 are each selected from: 、 、 、 、 and .如請求項1之化合物，其中M¹-L¹-T¹由以下表示：其中p2及p3各自為4、5或6。The compound of claim 1, wherein M¹ -L¹ -T¹ is represented by: wherein p2 and p3 are 4, 5 or 6 respectively.如請求項1或10之化合物，其中M²-L²-T²由以下表示：其中p2及p3各自為4、5或6。The compound of claim 1 or 10, wherein M² -L² -T² is represented by: wherein p2 and p3 are 4, 5 or 6 respectively.如請求項1之化合物，其中M¹-L¹-T¹及M²-L²-T²各自選自:、、、、、、、、、、、、、、、、及。The compound of claim 1, wherein M¹ -L¹ -T¹ and M² -L² -T² are each selected from: 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 and .如請求項1至12中任一項之化合物，其中-L³-G¹為-(CH₂)₃-OH或-(CH₂)₄-OH。The compound of any one of claims 1 to 12, wherein -L³ -G¹ is -(CH₂ )₃ -OH or -(CH₂ )₄ -OH.如請求項1之化合物，其中該化合物由式II-1表示：II-1 其中*表示立體純碳原子。The compound of claim 1, wherein the compound is represented by formula II-1: II-1 Where * represents a stereopure carbon atom.如請求項1至14中任一項之化合物，其中該化合物之特徵在於具有約15,000與30,000之間的維納路徑(Wiener path)及-15與0之間的MMFF94能量。The compound of any one of claims 1 to 14, wherein the compound is characterized by having a Wiener path between about 15,000 and 30,000 and an MMFF94 energy between -15 and 0.如請求項1至15中任一項之化合物，其中該化合物之特徵在於具有0.1與0.2之間的k。The compound of any one of claims 1 to 15, wherein the compound is characterized in having k between 0.1 and 0.2.如請求項1之化合物，其中該化合物選自表1。The compound of claim 1, wherein the compound is selected from Table 1.一種醫藥組合物，其包含如請求項1至17中任一項之化合物及核酸。A pharmaceutical composition comprising the compound of any one of claims 1 to 17 and a nucleic acid.如請求項18之醫藥組合物，其進一步包含以下中之一或多者：中性脂質、聚合物偶聯脂質、類固醇或陰離子兩親物。The pharmaceutical composition of claim 18, further comprising one or more of the following: a neutral lipid, a polymer-conjugated lipid, a steroid, or an anionic amphiphile.如請求項19之醫藥組合物，其進一步包含中性脂質。The pharmaceutical composition of claim 19, further comprising a neutral lipid.如請求項19或20之醫藥組合物，其中該中性脂質為磷脂。The pharmaceutical composition of claim 19 or 20, wherein the neutral lipid is a phospholipid.如請求項21之醫藥組合物，其中該磷脂係或包含二硬脂醯基磷脂醯膽鹼(DSPC)、二油醯基磷脂醯膽鹼(DOPC)、二肉豆蔻醯基磷脂醯膽鹼(DMPC)、雙十五醯基磷脂醯膽鹼、二月桂醯基磷脂醯膽鹼、二棕櫚醯基磷脂醯膽鹼(DPPC)、二花生醯基磷脂醯膽鹼(DAPC)、二山崳醯基磷脂醯膽鹼(DBPC)、雙二十三醯基磷脂醯膽鹼(DTPC)、雙二十四醯基磷脂醯膽鹼(DLPC)、棕櫚醯基油醯基-磷脂醯膽鹼(POPC)、1,2-二-O-十八烯基-sn-甘油-3-磷酸膽鹼(18:0二醚PC)、1-油醯基-2-膽固醇基半琥珀醯基-sn-甘油-3-磷酸膽鹼(OChemsPC)、1-十六基-sn-甘油-3-磷酸膽鹼(C16 Lyso PC)、二油醯基磷脂醯乙醇胺(DOPE)、二硬脂醯基-磷脂醯乙醇胺(DSPE)、二棕櫚醯基-磷脂醯乙醇胺(DPPE)、二肉豆蔻醯基-磷脂醯乙醇胺(DMPE)、二月桂醯基-磷脂醯乙醇胺(DLPE)、二植醯基-磷脂醯乙醇胺(DPyPE)、1,2-二-(9Z-十八碳烯醯基)-sn-甘油-3-磷酸膽鹼(DOPG)、1,2-二棕櫚醯基-sn-甘油-3-磷酸-(1′-rac-甘油) (DPPG)、1-棕櫚醯基-2-油醯基-sn-甘油-3-磷酸乙醇胺(POPE)或N-棕櫚醯基-D-赤-鞘胺醯基磷酸膽鹼(SM)。The pharmaceutical composition of claim 21, wherein the phospholipid is or comprises distearylphosphatidylcholine (DSPC), dioleoylphosphatidylcholine (DOPC), dimyristoylphosphatidylcholine (DMPC), dipentadecanoylphosphatidylcholine, dilaurylphosphatidylcholine, dipalmitoylphosphatidylcholine (DPPC), diarachidylphosphatidylcholine (DAPC), dibehenylphosphatidylcholine (DBPC), dioctadecanoylphosphatidylcholine (D ... Acylphosphatidylcholine (DTPC), di-tetracosylphosphatidylcholine (DLPC), palmitoyloleyl-phosphatidylcholine (POPC), 1,2-di-O-octadecenyl-sn-glycero-3-phosphocholine (18:0 diether PC), 1-oleyl-2-cholestyryl hemisuccinyl-sn-glycero-3-phosphocholine (OChemsPC), 1-hexadecyl-sn-glycero-3-phosphocholine (C16 Lyso PC), dioleylphosphatidylethanolamine (DOPE), distearylphosphatidylethanolamine (DSPE), dimalmitoylphosphatidylethanolamine (DPPE), dimyristoylphosphatidylethanolamine (DMPE), dilaurylphosphatidylethanolamine (DLPE), diphytylphosphatidylethanolamine (DPyPE), 1,2-bis-(9Z-octadecenyl)-sn-glycero-3-phosphocholine (DOPG), 1,2-dipalmitoyl-sn-glycero-3-phospho-(1′-rac-glycerol) (DPPG), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine (POPE), or N-palmitoyl-D-erythro-sphingamidoylphosphocholine (SM).如請求項19至22中任一項之醫藥組合物，其進一步包含聚合物偶聯脂質。The pharmaceutical composition of any one of claims 19 to 22, further comprising a polymer-coupled lipid.如請求項23之醫藥組合物，其中該聚合物偶聯脂質為包含聚合物之脂質，該聚合物為聚乙二醇。The pharmaceutical composition of claim 23, wherein the polymer-conjugated lipid is a lipid comprising a polymer, and the polymer is polyethylene glycol.如請求項24之醫藥組合物，其中該聚合物偶聯脂質選自PEG-DAG、PEG-PE、PEG-S-DAG、PEG2000-DMG、PEG-S-DMG、PEG-cer、PEG二烷氧基丙基胺基甲酸酯(例如，ω-甲氧基(聚乙氧基)乙基-N-(2,3-二(十四烷氧基)丙基)胺基甲酸酯或2,3-二(十四烷氧基)丙基-N-(ω-甲氧基(聚乙氧基)乙基)胺基甲酸酯)及其組合。The pharmaceutical composition of claim 24, wherein the polymer-coupled lipid is selected from PEG-DAG, PEG-PE, PEG-S-DAG, PEG2000-DMG, PEG-S-DMG, PEG-cer, PEG dialkoxypropylcarbamate (e.g., ω-methoxy(polyethoxy)ethyl-N-(2,3-di(tetradecyloxy)propyl)carbamate or 2,3-di(tetradecyloxy)propyl-N-(ω-methoxy(polyethoxy)ethyl)carbamate) and combinations thereof.如請求項19至25中任一項之醫藥組合物，其進一步包含類固醇。The pharmaceutical composition of any one of claims 19 to 25, further comprising a steroid.如請求項26之醫藥組合物，其中該類固醇為固醇。The pharmaceutical composition of claim 26, wherein the steroid is a sterol.如請求項27之醫藥組合物，其中該固醇為膽固醇。The pharmaceutical composition of claim 27, wherein the sterol is cholesterol.如請求項19至28中任一項之醫藥組合物，其進一步包含陰離子兩親物。The pharmaceutical composition of any one of claims 19 to 28, further comprising an anionic amphiphile.如請求項29之醫藥組合物，其中該陰離子兩親物選自DMGS、DPGS及DMGS。The pharmaceutical composition of claim 29, wherein the anionic amphiphile is selected from DMGS, DPGS and DMGS.如請求項19至22及26至29中任一項之醫藥組合物，其中該醫藥組合物包含中性脂質、類固醇及陰離子兩親物，且該醫藥組合物不包括聚合物偶聯脂質。The pharmaceutical composition of any one of claims 19 to 22 and 26 to 29, wherein the pharmaceutical composition comprises a neutral lipid, a steroid and an anionic amphiphile, and the pharmaceutical composition does not include a polymer-coupled lipid.如請求項18至31中任一項之醫藥組合物，其中該核酸為RNA。The pharmaceutical composition of any one of claims 18 to 31, wherein the nucleic acid is RNA.如請求項32之醫藥組合物，其中該RNA為modRNA、saRNA、taRNA或uRNA。The pharmaceutical composition of claim 32, wherein the RNA is modRNA, saRNA, taRNA or uRNA.如請求項18至31中任一項之醫藥組合物，其中該核酸為DNA。The pharmaceutical composition of any one of claims 18 to 31, wherein the nucleic acid is DNA.如請求項18至31中任一項之醫藥組合物，其中該核酸為RNA及DNA之混合物。The pharmaceutical composition of any one of claims 18 to 31, wherein the nucleic acid is a mixture of RNA and DNA.如請求項19至35中任一項之醫藥組合物，其中該醫藥組合物包含： 約30 mol%至約60 mol%之該化合物； 約20 mol%至約60 mol%之該類固醇； 約1 mol%至約2 mol%之該聚合物偶聯脂質；及 約5 mol%至約20 mol%之該中性脂質。The pharmaceutical composition of any one of claims 19 to 35, wherein the pharmaceutical composition comprises:about 30 mol% to about 60 mol% of the compound;about 20 mol% to about 60 mol% of the steroid;about 1 mol% to about 2 mol% of the polymer-conjugated lipid; andabout 5 mol% to about 20 mol% of the neutral lipid.如請求項19至35中任一項之醫藥組合物，其中該醫藥組合物包含： 約30 mol%至約60 mol%之該化合物； 約20 mol%至約60 mol%之該類固醇； 約0.5至約10 mol%之該陰離子兩親物；及 約5 mol%至約20 mol%之該中性脂質。The pharmaceutical composition of any one of claims 19 to 35, wherein the pharmaceutical composition comprises:about 30 mol% to about 60 mol% of the compound;about 20 mol% to about 60 mol% of the steroid;about 0.5 to about 10 mol% of the anionic amphiphile; andabout 5 mol% to about 20 mol% of the neutral lipid.如請求項37之醫藥組合物，其中該醫藥組合物不包括聚合物偶聯脂質。The pharmaceutical composition of claim 37, wherein the pharmaceutical composition does not include a polymer-coupled lipid.一種懸浮液，其包含分散相及水相，且其中該分散相包含呈粒子形式之如請求項18至38中任一項之醫藥組合物。A suspension comprising a dispersed phase and an aqueous phase, wherein the dispersed phase comprises the pharmaceutical composition of any one of claims 18 to 38 in the form of particles.如請求項39之懸浮液，其中該等粒子之平均直徑介於約50 nm至約200 nm之間。The suspension of claim 39, wherein the particles have an average diameter ranging from about 50 nm to about 200 nm.如請求項40之懸浮液，其中該水相包含該懸浮液中核酸之總濃度的小於20%。The suspension of claim 40, wherein the aqueous phase comprises less than 20% of the total concentration of nucleic acids in the suspension.如請求項39至41中任一項之懸浮液，其中該水相實質上不含該核酸。The suspension of any one of claims 39 to 41, wherein the aqueous phase is substantially free of the nucleic acid.一種治療疾病、病症或疾患之方法，其包含向個體投與如請求項39至42中任一項之懸浮液。A method of treating a disease, disorder or condition comprising administering to a subject the suspension of any one of claims 39 to 42.如請求項43之方法，其中該疾病、病症或疾患選自傳染病、癌症、遺傳病症、自體免疫疾病或罕見疾病。The method of claim 43, wherein the disease, disorder or condition is selected from an infectious disease, cancer, a genetic disorder, an autoimmune disease or a rare disease.一種增加個體之標靶中RNA表現或引起其增加之方法，其包含向該個體投與如請求項39至42中任一項之懸浮液。A method of increasing or causing an increase in RNA expression in a target in a subject, comprising administering to the subject the suspension of any one of claims 39 to 42.如請求項45之方法，其中該標靶選自肺、肝臟、脾、心臟、腦、淋巴結、膀胱、腎臟及胰臟。The method of claim 45, wherein the target is selected from the group consisting of lung, liver, spleen, heart, brain, lymph node, bladder, kidney, and pancreas.如請求項43至46中任一項之方法，其中該懸浮液係肌內、鼻內、靜脈內、皮下或腫瘤內投與。The method of any one of claims 43 to 46, wherein the suspension is administered intramuscularly, intranasally, intravenously, subcutaneously, or intratumorally.一種製備式III化合物，III 或其醫藥學上可接受之鹽之方法，該方法包含使式IV-1化合物與式IV-2化合物在熱存在下接觸：以提供第一中間產物，且其中使該第一中間產物與鉑觸媒及第一酸在氫氣存在下接觸以提供第二中間產物，且其中使該第二中間產物與第二酸接觸以提供該式III化合物，其中 M^1’和M^2’獨立地選自鍵或視情況經取代之C₂-C₁₀脂族基； L^1'及L^2'各自獨立地選自鍵、-OC(O)-及-C(O)O-； T^1'及T^2'各自獨立地選自視情況經取代之C₂-C₂₀脂族基； L^3’為視情況經取代之C₁-C₆脂族基； LG¹為適宜之脫離基；且 PG為適宜之醇保護基。A method for preparing a compound of formula III, III or a pharmaceutically acceptable salt thereof, which comprises contacting a compound of formula IV-1 with a compound of formula IV-2 in the presence of heat: to provide a first intermediate product, and wherein the first intermediate product is contacted with a platinum catalyst and a first acid in the presence of hydrogen to provide a second intermediate product, and wherein the second intermediate product is contacted with a second acid to provide the compound of formula III, wherein M^1' and M^2' are independently selected from a bond or an optionally substituted C₂ -C₁₀ aliphatic group; L^{1 '} and L^{2 '} are each independently selected from a bond, -OC(O)-, and -C(O)O-; T^{1 '} and T^{2 '} are each independently selected from an optionally substituted C₂ -C₂₀ aliphatic group; L^3' is an optionally substituted C₁ -C₆ aliphatic group; LG¹ is a suitable leaving group; and PG is a suitable alcohol protecting group.如請求項48之方法，其中式IV-2化合物藉由以下方式製備：使式V-1a及式V-1b化合物與式V-2化合物在鈀偶合觸媒、銅(I)輔觸媒及胺鹼存在下接觸，以提供該式IV-2化合物，其中LG²為適宜之脫離基。The method of claim 48, wherein the compound of formula IV-2 is prepared by: reacting the compounds of formula V-1a and formula V-1b with the compound of formula V-2 The reaction is carried out in the presence of a palladium coupling catalyst, a copper (I) cocatalyst and an amine base to provide the compound of formula IV-2, wherein LG² is a suitable detached group.一種製備式III-1化合物，III-1 或其醫藥學上可接受之鹽之方法，該方法包含使式IV-1化合物與式IV-3化合物在熱存在下接觸：以提供第一中間產物，且其中使該第一中間產物與酸接觸，以提供該式III-1化合物，其中 M^1’和M^2’各自獨立地選自鍵或視情況經取代之C₂-C₁₀脂族基； T^1’及T^2’各自獨立地選自視情況經取代之C₂-C₂₀脂族基； L^3’為視情況經取代之C₁-C₆脂族基； LG¹為適宜之脫離基；且 PG為適宜之醇保護基。A method for preparing a compound of formula III-1, III-1 or a pharmaceutically acceptable salt thereof, the method comprising contacting a compound of formula IV-1 with a compound of formula IV-3 in the presence of heat: to provide a first intermediate product, and wherein the first intermediate product is contacted with an acid to provide the compound of formula III-1, wherein M^1' and M^2' are each independently selected from a bond or an optionally substituted C₂ -C₁₀ aliphatic group; T^1' and T^2' are each independently selected from an optionally substituted C₂ -C₂₀ aliphatic group; L^3' is an optionally substituted C₁ -C₆ aliphatic group; LG¹ is a suitable leaving group; and PG is a suitable alcohol protecting group.