Detailed Description
Further advantages and effects of the present application will become readily apparent to those skilled in the art from the present disclosure, by describing embodiments of the present application with specific examples.
Definition of terms
The term "substituent" or "substituent group" refers to an atom or group of atoms that replaces a hydrogen atom on a molecule. The term "substituted" refers to a particular molecule bearing one or more substituents.
The term "alkyl" refers to branched and straight chain saturated aliphatic hydrocarbon groups containing, for example, 1 to 12 carbon atoms, 1 to 6 carbon atoms, or 1 to 4 carbon atoms. Examples of alkyl groups include, but are not limited to, methyl (Me), ethyl (Et), propyl (e.g., n-propyl and isopropyl), butyl (e.g., n-butyl, isobutyl, sec-butyl and tert-butyl), and pentyl (e.g., n-pentyl, isopentyl, neopentyl), n-hexyl, 2-methylpentyl, 2-ethylbutyl, 3-methylpentyl and 4-methylpentyl. When a number appears after the symbol "C" in the form of a subscript, the subscript more specifically defines the number of carbon atoms that a particular group may contain. For example, "C1-6 alkyl" represents straight and branched alkyl groups having one to six carbon atoms. Examples of C1-6 alkyl groups include methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl and pentyl.
The term "cycloalkyl" as used herein refers to a group derived from a non-aromatic monocyclic hydrocarbon molecule by removal of one hydrogen atom from a saturated ring carbon atom. Representative examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclopentyl, and cyclohexyl. When a numerical subscript appears after the symbol "C," the subscript more specifically defines the number of carbon atoms that a particular cycloalkyl group may contain. For example, "C3-6 cycloalkyl" represents a cycloalkyl group having three to six carbon atoms. Examples of monocyclic cycloalkyl groups are cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl or cycloheptyl.
The term "heteroatom" refers to oxygen (O), sulfur (S) and nitrogen (N).
The terms "halo" and "halogen" refer to F, cl, br and I.
The term "cyano" refers to the group-CN.
The term "amino" refers to the group-NH 2.
The term "heterocycloalkyl" or "heterocyclyl" refers to a non-aromatic 3-8 membered monocyclic, 7-12 membered bicyclic or 10-14 membered tricyclic ring system containing 1-4 heteroatoms (if monocyclic), 1-6 heteroatoms (if bicyclic) or 1-9 heteroatoms (if tricyclic) selected from O, S or N, wherein the non-aromatic ring system is fully saturated. The heterocycloalkyl group can be optionally substituted with one or more substituents. In one embodiment, 0,1, 2, 3 or 4 atoms of each ring of the heterocycloalkyl group may be substituted with substituents.
The term "aromatic ring" or "aryl" refers to a monocyclic, bicyclic, or tricyclic aromatic ring system of a hydrocarbon. The aryl group may be optionally substituted with one or more substituents. In one embodiment, 0, 1, 2, 3, 4, 5, or 6 atoms of each ring of the aryl group may be substituted with a substituent. Examples of aryl groups include phenyl, naphthyl, anthracenyl, fluorenyl, indenyl, azulenyl, and the like.
The term "aromatic heterocycle" or "heteroaryl" refers to substituted and unsubstituted aromatic 5 or 6 membered monocyclic groups and 9 or 10 membered bicyclic groups having at least one heteroatom (O, S or N) in at least one ring, preferably 1,2 or 3 heteroatoms independently selected from O, S and/or N. Each ring in a heteroaryl group containing a heteroatom may contain 1 or 2 oxygen or sulfur atoms and/or 1-4 nitrogen atoms provided that the total number of heteroatoms in each ring is 4 or less and that each ring has at least one carbon atom. The heteroaryl group may be attached to any available nitrogen or carbon atom of any ring. The heteroaryl ring system may be unsubstituted or may contain one or more substituents.
The term "alkoxy" refers to an-O-alkyl group. The alkoxy group may be optionally substituted with one or more substituents.
The term "isomer" or "stereoisomer" refers to a compound having the same chemical composition but a different arrangement of atoms or groups in space.
The term "pharmaceutically acceptable" refers to compounds, materials, compositions, and/or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit/risk ratio.
The compounds of formula I may form salts which are also within the scope of the present application. Unless otherwise indicated, references to a compound of the present application should be understood to include references to one or more salts thereof. The term "salt" means an acidic and/or basic salt formed with an inorganic and/or organic acid and a base. Furthermore, the term "salt" may include zwitterionic (inner salts), for example, when the compounds of formula I contain basic moieties (e.g. amine or pyridine or imidazole rings) as well as acidic moieties (e.g. carboxylic acids). Pharmaceutically acceptable (i.e., non-toxic, physiologically acceptable) salts are preferred, such as acceptable metal salts and amine salts, wherein the cation does not significantly contribute to the toxicity or biological activity of the salt. However, other salts may also be used, for example, in isolation or purification steps that may be employed during preparation, and are thus within the scope of the present application. Salts of the compounds of formula I may be formed, for example, by reacting a compound of formula I with an amount (e.g., one equivalent) of an acid or base in a medium such as one in which the salt is precipitated or in an aqueous medium, and then lyophilizing.
It is further understood that solvates (e.g., hydrates) of the compounds of formula I are also within the scope of the present application. The term "solvate" refers to a physical association of a compound of formula I with one or more solvent molecules (whether organic or inorganic). This physical association includes hydrogen bonding. In some cases, the solvate will be capable of isolation, for example, when one or more solvent molecules are incorporated into the crystal lattice of a crystalline solid. Exemplary solvates include hydrates, ethanolates, methanolates, isopropanolates, acetonitrile solvates and ethyl acetate solvates. Solvation methods are known in the art.
The term "metabolite" refers to a product produced by metabolizing a specific compound or salt thereof in vivo. Metabolites of compounds can be identified using conventional techniques known in the art and their activity determined using those techniques described herein. This product may result, for example, from oxidation, hydroxylation, reduction, hydrolysis, amidation, deamidation, esterification, deesterification, cleavage, etc. of the administered compound. Accordingly, the present application includes metabolites of the compounds of the present application, including compounds produced by a method comprising contacting a compound of the present application with a mammal for a period of time sufficient to produce a metabolite thereof.
The term "prodrug" or "prodrug" generally refers to a compound that is a prodrug that undergoes a chemical conversion by a metabolic or chemical process upon administration to a subject to yield a compound of formula I or a salt thereof. The content of prodrugs is well known in the art (see, e.g., berge et al (1977) "Pharmaceutical Salts", J.Pharm. Sci.66:1-19).
Examples of prodrug moieties include substituted and unsubstituted, branched or unbranched lower alkyl ester moieties (e.g., propionate), lower alkenyl esters, di-lower alkyl-amino lower alkyl esters (e.g., dimethylaminoethyl esters), amido lower alkyl esters (e.g., acetoxymethyl esters), acyloxy lower alkyl esters (e.g., pivaloyloxymethyl esters), aryl esters (phenyl esters), aryl-lower alkyl esters (e.g., benzyl esters), substituted (e.g., substituted with methyl, halogen or methoxy substituents) aryl and aryl-lower alkyl esters, amides, lower alkylamides, di-lower alkylamides, and hydroxyamides. Prodrugs that are converted to the active form by other mechanisms in the body are also included.
The compounds of formula I may form esters, which are also within the scope of the present application. The term "esters" refers to those esters that hydrolyze in vivo and include those that readily decompose in the human body to leave the parent compound or salt thereof. Suitable ester groups include, for example, those derived from pharmaceutically acceptable aliphatic carboxylic acids. Representative examples of specific esters include, but are not limited to, formate, acetate, propionate, butyrate, acrylate, and ethyl succinate.
Furthermore, the compounds of formula I may be isolated and purified after their preparation to give compositions containing the compounds of formula I in an amount equal to or greater than 99% by weight ("substantially pure") which are then used or formulated as described herein. Such "substantially pure" compounds of formula I are also considered to be part of the present application.
The compounds of the present application are intended to include all isotopes of atoms occurring in the compounds of the present application. Isotopes include atoms having the same atomic number but different mass numbers. By way of general example and not limitation, isotopes of hydrogen include deuterium (D) and tritium (T). Isotopes of carbon include13 C and14 C. Isotopically-labeled compounds of the present application can generally be prepared by conventional techniques known to those skilled in the art or by using an appropriate isotopically-labeled reagent in place of an unlabeled reagent employed in other instances by processes analogous to those described herein.
The term "derivative" refers to a compound formed by substitution of an atom or group of atoms in the parent compound molecule with other atoms or groups of atoms, referred to as a derivative of the parent compound. Such as halogenated hydrocarbons, alcohols, aldehydes, carboxylic acids can be considered derivatives of hydrocarbons, as they are products in which the hydrogen atom of a hydrocarbon is replaced with a halogen, hydroxy, oxygen, etc. Also, for example, acid halides, anhydrides, esters are carboxylic acid derivatives, as they are the products of substitution of hydroxyl groups in carboxylic acids with halogens and some organic groups. For example, when methane (CH4) is used as a parent, methanol (CH3 OH), formic acid (HCOOH), chloromethane (CH3 Cl) and the like are derivatives of methane.
The term "carrier" includes pharmaceutically acceptable carriers, excipients or stabilizers which are nontoxic to the cells or mammals at the dosages and concentrations employed. The physiologically acceptable carrier is typically an aqueous pH buffered solution. Non-limiting examples of physiologically acceptable carriers include buffers such as phosphates, citrates and other organic acids, antioxidants including ascorbic acid, low molecular weight (less than about 10 residues) polypeptides, proteins such as serum albumin, gelatin or immunoglobulins, hydrophilic polymers such as polyvinylpyrrolidone, amino acids such as glycine, glutamine, asparagine, arginine or lysine, monosaccharides, disaccharides and other carbohydrates including glucose, mannose or dextrins, chelating agents such as EDTA, sugar alcohols such as mannitol or sorbitol, salt forming counter ions such as sodium, and/or nonionic surfactants such as TWEENTM, polyethylene glycol (PEG) and PLURONICSTM. In certain embodiments, the pharmaceutically acceptable carrier is a non-naturally occurring pharmaceutically acceptable carrier.
The term "inhibit" means to reduce the activity of the enzyme of interest compared to the activity of the enzyme in the absence of the inhibitor. In some embodiments, the term "inhibit" means a reduction in HPK1 activity of at least about 5%, at least about 10%, at least about 20%, at least about 25%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or at least about 95%. In other embodiments, inhibition means a reduction in HPK1 activity of about 5% to about 25%, about 25% to about 50%, about 50% to about 75%, or about 75% to 100%. In some embodiments, inhibition means about a 95% to 100% reduction in HPK1 activity, e.g., a 95%, 96%, 97%, 98%, 99% or 100% reduction in activity. This decrease can be measured using a variety of techniques, which are well known to those skilled in the art, including in vitro kinase assays.
An "HPK1 antagonist" or "HPK1 inhibitor" is a molecule that reduces, inhibits, or otherwise reduces one or more biological activities of HPK1 (e.g., serine/threonine kinase activity, recruitment to the TCR complex upon TCR activation, interaction with a protein binding partner (e.g., SLP 76)). Antagonism using the HPK1 antagonists does not necessarily indicate complete elimination of HPK1 activity. In contrast, the activity can be reduced by a statistically significant amount, including, for example, reducing HPK1 activity by at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 95%, or 100% as compared to an appropriate control.
Detailed Description
Quinazoline compounds
In one aspect, the application provides compounds of formula I:
Or a pharmaceutically acceptable salt, solvate, stereoisomer, prodrug, metabolite, or derivative thereof, wherein one of Ra,Rb,Rc,Rd may beThe remainder may each be independently selected from H, hydroxy, halogen, cyano, C1-6 alkyl, C3-8 cycloalkyl, C1-6 substituted alkyl, C3-8 substituted cycloalkyl, and C1-6 alkoxy;
Re can be selected from H, hydroxy, halogen, cyano, C1-6 alkyl, C3-8 cycloalkyl, C1-6 substituted alkyl, C3-8 substituted cycloalkyl, and C1-6 alkoxy;
X is selected from the group consisting of-O- (O) -NH- (NH) -and S-, SO-, -SO2 -, carbonyl-, carbonylamino-and aminocarbonyl;
R1 can be substituted cycloalkyl, substituted heterocyclyl orM may be 0,1, 2, 3, 4 or 5, and r14 may be C1-6 alkyl, C3-8 cycloalkyl, C1-6 substituted alkyl or C3-8 substituted cycloalkyl;
the A ring may be a substituted benzene ring or a substituted 5-6 membered aromatic heterocyclic ring, wherein the substituted 5-6 membered aromatic heterocyclic ring has 1-4 heteroatoms selected from O, S and N;
R2 may be selected from the following substituents:
Wherein Y may be C or N, R7 may be H, C1-6 alkyl, C3-8 cycloalkyl, C1-6 substituted alkyl or C3-8 substituted cycloalkyl, R8 may be H, c1-6 alkyl, C3-8 cycloalkyl, C1-6 substituted alkyl or C3-8 substituted cycloalkyl, R9 and R10 can independently be H, C1-6 alkyl, C3-8 cycloalkyl, C1-6 substituted alkyl or C3-8 substituted cycloalkyl, R11 can be H, Amino, C1-6 alkyl, C3-8 cycloalkyl, C1-6 substituted alkyl or C3-8 substituted cycloalkyl, R12 can be a 4-6 membered heterocyclyl or a 4-6 substituted heterocyclyl.
In the present application, one of Ra,Rb,Rc,Rd may beThe remainder may each independently be selected from H, hydroxy, halogen, cyano, C1-6 alkyl, C3-8 cycloalkyl, C1-6 substituted alkyl, C3-8 substituted cycloalkyl and C1-6 alkoxy represent a structure when one of Ra,Rb,Rc,Rd isWhen the remaining three may each be independently selected from H, hydroxy, halogen, cyano, C1-6 alkyl, C3-8 cycloalkyl, C1-6 substituted alkyl, C3-8 substituted cycloalkyl and C1-6 alkoxy, e.g., when Ra isWhen Rb,Rc,Rd may each be independently selected from H, hydroxy, halogen, cyano, C1-6 alkyl, C3-8 cycloalkyl, C1-6 substituted alkyl, C3-8 substituted cycloalkyl, and C1-6 alkoxy.
In some embodiments, the R1 may beWhen m takes on a value of 1 or 2.
In some embodiments, the R14 can be C1-6 alkyl.
In some embodiments, the R1 may be a substituted 5-8 membered cycloalkyl or a substituted 5-8 membered heterocyclyl, wherein the substituted 5-8 membered heterocyclyl has 1-4 heteroatoms selected from O, S and N.
In some embodiments, the R1 may be a substituted 5-6 membered cycloalkyl or a substituted 5-6 membered heterocyclyl, wherein the substituted 5-6 membered heterocyclyl has 1-4 heteroatoms selected from O, S and N.
In some embodiments, when R1 is a substituted 5-8 membered cycloalkyl, the R1 may beWherein n may be 1, 2, 3 or 4.
In some embodiments, the R3 may be-OR4 OR-NR5R6, where R4 may be H,Alkyl, substituted alkyl, cycloalkyl or substituted cycloalkyl, R5 and R6 may independently be H,Alkyl, substituted alkyl, cycloalkyl or substituted cycloalkyl, wherein R5 and R6 are not both cycloalkyl or substituted cycloalkyl.
In some embodiments, the R4 can be H, C1-6 alkyl, C1-6 substituted alkyl, C3-8 cycloalkyl, or C3-8 substituted cycloalkyl.
For example, when R4 is H, R3 is-OH and R1 may beWherein n may be 1, 2, 3 or 4.
In some embodiments, the R5 and R6 may independently be H, C1-6 alkyl, C1-6 substituted alkyl, C3-8 cycloalkyl, or C3-8 substituted cycloalkyl, wherein R5 and R6 are not both C3-8 cycloalkyl or C3-8 substituted cycloalkyl.
For example, when R5 and R6 are both H, R3 is-NH2,R1 may beWherein n is 1, 2, 3 or 4.
In some embodiments, the R13 can be H, C1-6 alkyl, C1-6 substituted alkyl, C3-8 cycloalkyl, or C3-8 substituted cycloalkyl.
In some embodiments, when R4 isWhen R3 is
For example, when R13 is methyl, R3 isR1 isWherein n may be 1, 2, 3 or 4.
In some embodiments, when R5 is H, R6 isWhen R3 is
For example, when R13 is methyl, R3 isR1 isWherein n may be 1, 2, 3 or 4.
In some embodiments, R1 may be selected from the following substituents:
in some embodiments, the a ring may be selected from the following substituted aromatic rings:
In some embodiments, the a ring may be a substituted benzene ring, a substituted pyridine ring, or a substituted pyrazole ring.
In some embodiments, when the a ring is a substituted benzene ring or a substituted pyridine ring, the R2 substitution position may be at the para or meta position of X.
For example, the a ring may be selected from the following structures:
In some embodiments, R2 may be selected from the following substituents:
For example, the a ring may be selected from the following structures:
in some embodiments, the a ring may be selected from the following structures:
In some embodiments, R12 may be selected from the following substituents:
For example, the a ring may be selected from the following structures:
In some embodiments, the compound of formula I is as shown in formula I-1,
Or a pharmaceutically acceptable salt, solvate, stereoisomer, prodrug, metabolite, or derivative thereof, wherein Rb,Rc,Rd,Re can each be independently selected from H, hydroxy, halo, cyano, C1-6 alkyl, C3-8 cycloalkyl, C1-6 substituted alkyl, C3-8 substituted cycloalkyl, and C1-6 alkoxy;
X is selected from the group consisting of-O- (O) -NH- (NH) -and S-, SO-, -SO2 -, carbonyl-, carbonylamino-and aminocarbonyl;
R1 can be substituted cycloalkyl, substituted heterocyclyl orM may be 0, 1, 2, 3, 4 or 5, r14 may be C1-6 alkyl, C3-8 cycloalkyl, C1-6 substituted alkyl or C3-8 substituted cycloalkyl;
the A ring may be a substituted benzene ring or a substituted 5-6 membered aromatic heterocyclic ring, wherein the substituted 5-6 membered aromatic heterocyclic ring has 1-4 heteroatoms selected from O, S and N;
R2 may be selected from the following substituents:
Wherein Y may be C or N, R7 is H, C1-6 alkyl, C3-8 cycloalkyl, C1-6 substituted alkyl or C3-8 substituted cycloalkyl, R8 may be H, c1-6 alkyl, C3-8 cycloalkyl, c1-6 substituted alkyl or C3-8 substituted cycloalkyl, R9 and R10 can independently be H, C1-6 alkyl, C3-8 cycloalkyl, C1-6 substituted alkyl or C3-8 substituted cycloalkyl, R11 can be H, Amino, C1-6 alkyl, C3-8 cycloalkyl, C1-6 substituted alkyl or C3-8 substituted cycloalkyl, R12 can be a 4-6 membered substituted heterocyclic group or a 4-6 membered unsubstituted heterocyclic group.
In some embodiments, the compound of formula I may be as shown in formula I-1-1 or I-1-2,
Or a pharmaceutically acceptable salt, solvate, stereoisomer, prodrug, metabolite, or derivative thereof, wherein Rb,Rc,Rd,Re can each be independently selected from H, hydroxy, halo, cyano, C1-6 alkyl, C3-8 cycloalkyl, C1-6 substituted alkyl, C3-8 substituted cycloalkyl, and C1-6 alkoxy;
X is selected from the group consisting of-O- (O) -NH- (NH) -and S-, SO-, -SO2 -, carbonyl-, carbonylamino-and aminocarbonyl;
n may be 1,2, 3 or 4;
m may be 0, 1,2,3,4 or 5;
R14 can be C1-6 alkyl, C3-8 cycloalkyl, C1-6 substituted alkyl or C3-8 substituted cycloalkyl;
the A ring may be a substituted benzene ring or a substituted 5-6 membered aromatic heterocyclic ring, wherein the substituted 5-6 membered aromatic heterocyclic ring has 1-4 heteroatoms selected from O, S and N;
R2 may be selected from the following substituents:
Wherein Y may be C or N, R7 may be H, C1-6 alkyl, C3-8 cycloalkyl, C1-6 substituted alkyl or C3-8 substituted cycloalkyl, R8 may be H, c1-6 alkyl, C3-8 cycloalkyl, c1-6 substituted alkyl or C3-8 substituted cycloalkyl, R9 and R10 can independently be H, C1-6 alkyl, C3-8 cycloalkyl, C1-6 substituted alkyl or C3-8 substituted cycloalkyl, R11 can be H, Amino, C1-6 alkyl, C3-8 cycloalkyl, C1-6 substituted alkyl or C3-8 substituted cycloalkyl, R12 can be a 4-6 membered substituted heterocyclic group or a 4-6 membered unsubstituted heterocyclic group.
In some embodiments, the R3 may be-OR4 OR-NR5R6, where R4 may be H,Alkyl, substituted alkyl, cycloalkyl or substituted cycloalkyl, R5 and R6 may independently be H,Alkyl, substituted alkyl, cycloalkyl or substituted cycloalkyl, wherein R5 and R6 are not both cycloalkyl or substituted cycloalkyl.
In some embodiments, the R4 can be H, C1-6 alkyl, C1-6 substituted alkyl, C3-8 cycloalkyl, or C3-8 substituted cycloalkyl.
For example, when R4 is H, R3 is-OH and R1 may beWherein n is1, 2, 3 or 4, the compound of formula I can be shown as formula I-1-1a,
In some embodiments, the R5 and R6 may independently be H, C1-6 alkyl, C1-6 substituted alkyl, C3-8 cycloalkyl, or C3-8 substituted cycloalkyl, wherein R5 and R6 are not both C3-8 cycloalkyl or C3-8 substituted cycloalkyl.
For example, when R5 and R6 are both H, R3 is-NH2,R1 may beWherein n is 1,2, 3 or 4, the compound of formula I can be shown as formula I-1-1b,
In some embodiments, when R4 isWhen R3 isThe compound of the formula I can be shown as a formula I-1-1c, wherein n can be 1, 2, 3 or 4,
In some embodiments, the R13 can be H, C1-6 alkyl, C1-6 substituted alkyl, C3-8 cycloalkyl, or C3-8 substituted cycloalkyl.
For example, when R13 is methyl, R3 isR1 isWherein n may be 1, 2, 3 or 4.
In some embodiments, when R5 is H, R6 isWhen R3 isThe compound of the formula I can be shown as a formula I-1-1d, wherein n can be 1, 2, 3 or 4,
In some embodiments, the R13 can be H, C1-6 alkyl, C1-6 substituted alkyl, C3-8 cycloalkyl, or C3-8 substituted cycloalkyl.
For example, when R13 is methyl, R3 isR1 isWherein n may be 1, 2, 3 or 4.
In some embodiments, R1 may be selected from the following substituents:
in some embodiments, the a ring may be selected from the following substituted aromatic rings:
In some embodiments, the a ring may be a substituted benzene ring, a substituted pyridine ring, or a substituted pyrazole ring.
In some embodiments, when the a ring is a substituted benzene ring or a substituted pyridine ring, the R2 substitution position may be at the para or meta position of X.
For example, the a ring may be selected from the following structures:
In some embodiments, R2 may be selected from the following substituents:
For example, the a ring may be selected from the following structures:
in some embodiments, the a ring may be selected from the following structures:
In some embodiments, R12 may be selected from the following substituents:
For example, the a ring may be selected from the following structures:
In some embodiments, the compound of formula I may be selected from the following compounds:
In some embodiments, the compound of formula I may be as shown in formula I-2,
Or a pharmaceutically acceptable salt, solvate, stereoisomer, prodrug, metabolite, or derivative thereof, wherein Rb,Rc,Rd,Re can each be independently selected from H, hydroxy, halo, cyano, C1-6 alkyl, C3-8 cycloalkyl, C1-6 substituted alkyl, C3-8 substituted cycloalkyl, and C1-6 alkoxy;
X is selected from the group consisting of-O- (O) -NH- (NH) -and S-, SO-, -SO2 -, carbonyl-, carbonylamino-and aminocarbonyl;
R1 can be substituted cycloalkyl, substituted heterocyclyl orM may be 0,1, 2, 3, 4 or 5, and r14 may be C1-6 alkyl, C3-8 cycloalkyl, C1-6 substituted alkyl or C3-8 substituted cycloalkyl;
the A ring is a substituted benzene ring or a substituted 5-6 membered aromatic heterocyclic ring, wherein the substituted 5-6 membered aromatic heterocyclic ring has 1-4 heteroatoms selected from O, S and N;
R2 may be selected from the following substituents:
Wherein Y may be C or N, R7 may be H, C1-6 alkyl, C3-8 cycloalkyl, C1-6 substituted alkyl or C3-8 substituted cycloalkyl, R8 may be H, c1-6 alkyl, C3-8 cycloalkyl, c1-6 substituted alkyl or C3-8 substituted cycloalkyl, R9 and R10 can independently be H, C1-6 alkyl, C3-8 cycloalkyl, C1-6 substituted alkyl or C3-8 substituted cycloalkyl, R11 can be H, Amino, C1-6 alkyl, C3-8 cycloalkyl, C1-6 substituted alkyl or C3-8 substituted cycloalkyl, R12 can be a 4-6 membered substituted heterocyclic group or a 4-6 membered unsubstituted heterocyclic group.
In some embodiments, the compound of formula I may be as shown in formula I-3,
Or a pharmaceutically acceptable salt, solvate, stereoisomer, prodrug, metabolite, or derivative thereof, wherein Rb,Rc,Rd,Re can each be independently selected from H, hydroxy, halo, cyano, C1-6 alkyl, C3-8 cycloalkyl, C1-6 substituted alkyl, C3-8 substituted cycloalkyl, and C1-6 alkoxy;
X is selected from the group consisting of-O- (O) -NH- (NH) -and S-, SO-, -SO2 -, carbonyl-, carbonylamino-and aminocarbonyl;
R1 can be substituted cycloalkyl, substituted heterocyclyl orM may be 0,1, 2, 3, 4 or 5, and r14 may be C1-6 alkyl, C3-8 cycloalkyl, C1-6 substituted alkyl or C3-8 substituted cycloalkyl;
the A ring is a substituted benzene ring or a substituted 5-6 membered aromatic heterocyclic ring, wherein the substituted 5-6 membered aromatic heterocyclic ring has 1-4 heteroatoms selected from O, S and N;
R2 may be selected from the following substituents:
Wherein Y can be C or N and R7 can be H, C1-6 alkyl, C3-8 cycloalkyl, C1-6 substituted alkyl or C3-8 substituted cycloalkyl, R8 can be H, c1-6 alkyl, C3-8 cycloalkyl, c1-6 substituted alkyl or C3-8 substituted cycloalkyl, R9 and R10 can independently be H, C1-6 alkyl, C3-8 cycloalkyl, C1-6 substituted alkyl or C3-8 substituted cycloalkyl, R11 can be H, Amino, C1-6 alkyl, C3-8 cycloalkyl, C1-6 substituted alkyl or C3-8 substituted cycloalkyl, R12 can be a 4-6 membered substituted heterocyclic group or a 4-6 membered unsubstituted heterocyclic group.
In some embodiments, the compound of formula I may be as shown in formula I-4,
Or a pharmaceutically acceptable salt, solvate, stereoisomer, prodrug, metabolite, or derivative thereof, wherein Rb,Rc,Rd,Re can each be independently selected from H, hydroxy, halo, cyano, C1-6 alkyl, C3-8 cycloalkyl, C1-6 substituted alkyl, C3-8 substituted cycloalkyl, and C1-6 alkoxy;
X is selected from the group consisting of-O- (O) -NH- (NH) -and S-, SO-, -SO2 -, carbonyl-, carbonylamino-and aminocarbonyl;
R1 can be substituted cycloalkyl, substituted heterocyclyl orM may be 0,1, 2, 3, 4 or 5, and r14 may be C1-6 alkyl, C3-8 cycloalkyl, C1-6 substituted alkyl or C3-8 substituted cycloalkyl;
the A ring is a substituted benzene ring or a substituted 5-6 membered aromatic heterocyclic ring, wherein the substituted 5-6 membered aromatic heterocyclic ring has 1-4 heteroatoms selected from O, S and N;
R2 may be selected from the following substituents:
Wherein Y can be C or N and R7 can be H, C1-6 alkyl, C3-8 cycloalkyl, C1-6 substituted alkyl or C3-8 substituted cycloalkyl, R8 can be H, c1-6 alkyl, C3-8 cycloalkyl, c1-6 substituted alkyl or C3-8 substituted cycloalkyl, R9 and R10 can independently be H, C1-6 alkyl, C3-8 cycloalkyl, C1-6 substituted alkyl or C3-8 substituted cycloalkyl, R11 can be H, Amino, C1-6 alkyl, C3-8 cycloalkyl, C1-6 substituted alkyl or C3-8 substituted cycloalkyl, R12 can be a 4-6 membered substituted heterocyclic group or a 4-6 membered unsubstituted heterocyclic group.
Pharmaceutical composition
In another aspect, the present application provides a pharmaceutical composition comprising at least one compound of formula I, or a pharmaceutically acceptable salt, solvate, stereoisomer, prodrug, metabolite, or derivative thereof, and one or more non-toxic pharmaceutically acceptable carriers and/or diluents and/or adjuvants (collectively referred to herein as "carrier" materials), and, when desired, other active ingredients.
Without intending to be limited by any theory, the following examples are meant to illustrate the compounds, methods of preparation, uses, and the like of the present application and are not intended to limit the scope of the application.
Examples
EXAMPLE 1 Synthesis of the Compound DD02001H 8- (((1 s,4 s) -4-aminocyclohexyl) oxo) -N- (1- (1-methylpiperidin-4-yl) -1H-pyrazol-4-yl) quinazolin-2-amine
FIG. 1 shows the synthetic route for compound DD 02001H.
1.1 Synthesis of Compound 2
To a solution of compound 1 (25.0 g,150mmol,1.00 eq) in THF (300 mL) was added BH3/THF (1.00 m,329mL,2.20 eq) dropwise under protection of N2 at 0 ℃. After the completion of the dropwise addition, the mixture was stirred at 0 ℃ for 30 minutes, and then the mixture was heated to 50 ℃ and stirred for 12 hours. TLC (petroleum ether: ethyl acetate=1:1, starting material Rf =0.1, product Rf =0.3) showed new spot formation, complete disappearance of starting material, complete reaction, subsequent cooling of the mixture to 0 ℃, dropwise addition of MeOH (400 mL) to no bubble formation, then addition of 40.0mL of H2 O and extraction with ethyl acetate (300 mL x 2), washing of the organic layer brine (100 mL x 2), drying and filtration over anhydrous Na2SO4, and concentration under reduced pressure gave compound 2 (45.0 g, 254 mmol,98.2% yield) as a pale yellow oily substance, structure confirmed by1 H NMR.
1H NMR(400MHz,CDCl3)δ6.65-6.85(m,3H),4.63(s,2H),3.86(s,3H)。
1.2 Synthesis of Compound 3
To a solution of compound 2 (45.0 g, 254 mmol,1.00 eq.) in Dichloromethane (DCM) (500 mL) was added MnO2 (128 g,1.47mol,5.00 eq.) and the resulting mixture was stirred at 25℃for 12 hours. Thin layer chromatography (petroleum ether: ethyl acetate=1:1, material Rf =0.45, product Rf =0.8) showed new spot formation, the starting material spot disappeared, the mixture was filtered through celite after the reaction was completed, and the filtrate was concentrated under reduced pressure. The compound 3 (25.0 g,166mmol,56.3% yield) was isolated and purified by column chromatography (SiO2, petroleum ether/ethyl acetate=30/1 to 5/1) and the structure confirmed by1 H NMR.
1H NMR(400MHz,DMSO-d6)δ9.85(s,1H),7.18(dd,J＝8.0,1.2Hz,1H),7.02(dd,J＝8.0,0.8Hz,1H),6.80(br s,2H),6.64(t,J＝8.0Hz,1H),3.77-3.87(m,3H).
1.3 Synthesis of Compound 4
A mixture of compound 3 (23.0 g,152mmol,1.00 eq), urea (101 g,1.67mol,89.7mL,11.0 eq) and NH4 OAc (586 mg,7.61mmol,0.05 eq) was stirred at 160℃for 0.5 h, the mixture began to precipitate from the hot solution. After NMP (100 mL) was added to dissolve the solid precipitate, the reaction was stirred at 160℃for 1 hour. LC-MS showed that MS (rt=0.247 min) was detected as target product 4, starting material disappeared, after the end of the reaction the mixture was cooled to 25 ℃ and poured into 100mL H2 O, stirred for 10 min at 25 ℃ and filtered under reduced pressure. The crude product was dispersed with petroleum ether (60.0 mL) at 25 ℃ with shaking for 5min and filtered under reduced pressure to give compound 4 (20.0 g,114mmol,74.6% yield) as a grey solid. The structure was confirmed by1 H NMR.
LCMS product rt=0.247 min, M/z=177.2 (m+h)+
1H NMR(400MHz,DMSO)δ8.36(s,1H),6.90(br d,J＝4.0Hz,1H),6.87(d,J＝8.0Hz,1H),6.79(dd,J＝8.0,4.0Hz,1H),5.98(dd,J＝8.0,4.0Hz,1H),3.79(s,3H).
1.4 Synthesis of Compound 5
Compound 4 (19.0 g,108mmol,1.00 eq) was added to POCl3 (248 g,1.61mol,150mL,15.0 eq) at 0℃and the resulting mixture stirred at 25℃for 0.5h, then warmed to 140℃and stirred for 2 h. LC-MS showed detection of the target product MS (rt=0.655 minutes). After the reaction was completed, the mixture was slowly poured into ice water (200 mL) at 0-10 ℃ and stirred, then extracted with ethyl acetate (300 mL x 4), the organic layer was washed with brine (100 mL x 2), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. Shaking dispersion with petroleum ether (60.0 mL) at 25℃for 5min, and filtration gave compound 5 (7.60 g,39.1mmol,36.2% yield) as a yellow solid, the structure was confirmed by1 H NMR.
LCMS product rt=0.655 min, M/z=195.0 (m+h)+
1H NMR(400 MHz,DMSO)δ9.56(s,1H),7.69-7.79(m,2H),7.54(br d,J=4.0 Hz,1H),3.99(s,3H)。
1.5 Synthesis of Compound 6
DIAD (26.3 g,130 mmol,25.3 mL,1.50 eq) was added to a solution of compound 6-1 (10.0 g,86.8 mmol,10.1 mL,1.00 eq), compound 6-1a (11.7 g,104mmol,1.20 eq), and triphenylphosphine (34.2 g,130 mmol,1.50 eq) in THF (300 mL) at 0deg.C and the resulting mixture was stirred under 25℃ C, N2 for 12 hours. LCMS showed complete reaction of starting material and product formation. After the reaction was completed, the reaction mixture was adjusted to ph=4 with 1M HCl and extracted with ethyl acetate (300 mL), the aqueous phase was adjusted to ph=8 with NaHCO3 (saturated) and extracted again with ethyl acetate (100 ml x 2). The organic layer was washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give compound 6-2 (8.00 g,38.1 mmol,43.8% yield) as a yellow oily substance, the structure of which was confirmed by1 H NMR.
LCMS product rt=0.150 min, M/z=211.1 (m+h)+
1H NMR(400 MHz,CDCl3)δ8.15(s,1H),8.04(s,1H),4.08-4.14(m,1H),2.95-2.98(m,2H),2.31(s,3H),2.09-2.15(m,6H).
Compound 6-2 (8.00 g,38.1 mmol,1.00 eq), pd/C (2.00 g,10% purity) was stirred in methanol (50.0 mL) and the mixture stirred at 25℃under H2 (15 psi) for 12 hours. LCMS showed complete consumption of compound 6-2, and after completion of the reaction the mixture was filtered and concentrated under reduced pressure to give compound 6 (5.00 g,27.7 mmol,72.9% yield) as a yellow oil, the structure confirmed by1 H NMR.
LCMS product rt=0.10 min, M/z= 181.1 (m+h)+
1H NMR(400 MHz,CDCl3)δ7.09(s,1H),7.01(s,1H),3.92-3.98(m,1H),2.88-2.91(m,4H),2.26(s,3H),1.85-2.26(m,6H).
1.6 Synthesis of Compound 7
To a solution of compound 5 (400 mg,2.06mmol,1.00 eq) and compound 6 (444 mg,2.47mmol,1.20 eq) in IPA (10.0 mL) was added TFA (23.4 mg,205umol,15.2ul,0.10 eq) and the resulting mixture was stirred at 100 ℃ for 2 hours. LCMS showed the detection of the target product mass (rt=0.653 min). The mixture was diluted with ethyl acetate (100 mL) and washed with saturated NaHCO3 solution (50.0 mL), the organic phase was washed with brine (50.0 mL), dried over anhydrous sodium sulfate, filtered and concentrated to give a solid material which was dispersed with petroleum ether (25.0 mL) with shaking to give compound 7 (570 mg,1.68mmol,81.9% yield) as a yellow solid, the structure was confirmed by LCMS and1 H NMR.
LCMS product rt=0.673 min, M/z= 339.3 (m+h)+
1H NMR(400MHz,CDCl3)δ9.05(s,1H),8.22(s,1H),7.60(s,1H),7.31-7.35(m,2H),7.21-7.25(m,1H),7.10-7.12(m,1H),4.12-4.14(m,1H),4.05(s,3H),2.98-3.01(m,2H),2.34(s,3H),2.14-2.23(m,6H).
1.7 Synthesis of Compound 8
To a solution of compound 7 (570 mg,1.68mmol,1.00 eq) in DCM (15.0 mL) was added BBr3 (1.05 g,4.21mmol,405.7uL,2.50 eq) dropwise. The mixture was stirred at 25 ℃ for 12 hours. LCMS (EW 20001-101-P1 A1) showed the detection of the target product (rt=0.622 min). At the end of the reaction, the mixture was diluted with DCM (100 mL) and washed with saturated NaHCO3 solution (50.0 mL), the organic phase was washed with brine (50.0 mL), dried over anhydrous sodium sulfate, filtered and concentrated to give compound 8 (500 mg, crude) as a yellow solid which was used directly in the next step.
LCMS product rt=0.847 min, M/z= 325.2 (m+h)+
1.8 Synthesis of Compound 10
To a solution of compound 8 (200 mg,616umol,1.00 eq) and compound 9 (273 mg,740umol,1.20 eq) in DMF (3.00 mL) was added Cs2CO3 (502 mg,1.54mmol,2.50 eq) and the resulting mixture was stirred at 80℃for 1 hour. LCMS showed the detection of the target product (rt=0.799 min). After the reaction was completed, the mixture was quenched with water (50.0 mL) and extracted with ethyl acetate (50.0 mL x 2), the organic phase was washed with brine (50.0 mL), dried over anhydrous sodium sulfate, filtered and concentrated to give compound 10 (320 mg, 803 umol,99.5% yield) as a yellow oily substance which was used directly in the next step without further purification.
LCMS product rt=0.799 min, M/z=522.3 (m+h)+.
1.9 Synthesis of Compound DD02001H
A mixture of compound 10 (320 mg, 313 umol,1.00 eq) and HCl/dioxane (4M, 10.0mL,65.2 eq) was stirred at 25℃for 1 hour. LCMS showed the detection of the target product (rt=0.651 min). The mixture was concentrated to give a residue which was purified by preparative HPLC. The mixture was adjusted to ph=8 with saturated NaHCO3 solution and extracted with DCM (50.0 mL x 2), the organic phase was washed with brine (50.0 mL), dried over anhydrous sodium sulfate, filtered and concentrated to give compound DD02001H, the structure of which was confirmed by LCMS, HPLC (EW 20001-106-P1 A2) and1 H-NMR as a yellow solid (27.2 mg,91.7umol,14.9% yield, 96.7% purity).
LCMS product rt=0.644 min, M/z=422.3 (m+h)+
1H NMR(400MHz,CDCl3)δ8.94(s,1H),8.14(s,1H),7.68(s,1H),7.23-7.25(m,1H),7.12-7.13(m,2H),7.06(s,1H),4.67-4.68(m,1H),4.05-4.09(m,1H),2.89-2.92(m,2H),2.78-2.80(m,1H),2.26(s,3H),2.05-2.15(m,8H),1.62-1.74(m,8H).
EXAMPLE 2 Synthesis of Compound DD02013H 4- ((2- ((4- ((2- (dimethylamino) ethyl) (methyl) amino) phenyl) amino) quinazolin-8-yl) oxy) cyclohexan-1-ol
FIG. 2 shows the synthetic route for compound DD 02013H.
2.1 Synthesis of Compound 11
To a solution of compound 5 (500 mg,2.57mmol,1.00 eq) in DCM (10.0 mL) was added dropwise a solution of BBr3 (1.42 g,5.65mmol,545uL,2.20 eq) in DCM (5.00 mL) at 0 ℃. After the completion of the dropwise addition, the mixture was stirred at 25℃for 12 hours. LCMS showed the detection of the target product molecular weight (rt=0.585 min). After the reaction was complete, the mixture was quenched with water (50.0 mL) and extracted with DCM (50.0 mL). The organic phase was washed with brine (50.0 mL), dried over anhydrous sodium sulfate, filtered and concentrated to give a residue which was purified by column chromatography (SiO2, petroleum ether/ethyl acetate=20/1 to 10/1, tlc (petroleum ether: ethyl acetate=3:1, rf =0.3) to give compound 11 (400 mg,2.21mmol,86.2% yield) as a yellow solid, the structure was confirmed by LCMS and1 H NMR.
LCMS product rt=0.575 min, M/z=181.0 (m+h)+
1H NMR(400MHz,CDCl3)δ9.30(s,1H),7.59-7.64(m,1H),7.49-7.52(m,1H),7.42-7.45(m,2H)。
2.2 Synthesis of Compound 12
To a solution of compound 11 (200 mg,1.11mmol,1.00 eq) and compound 11a (639 mg,1.66mmol,1.50 eq) in DMF (5.00 mL) was added Cs2CO3 (721.6 mg,2.21mmol,2.00 eq) and the resulting mixture was stirred at 80℃for 2 hours. LCMS detected the target product molecular weight (rt= 1.229 min). After the reaction was completed, the reaction was quenched by addition of water (50.0 mL), extracted with ethyl acetate (50.0 mL x 2), the organic phase was washed with brine (50.0 mL), dried over anhydrous sodium sulfate, filtered and concentrated to give a residue which was purified by column chromatography (SiO2, petroleum ether/ethyl acetate=20/1 to 10/1, tlc (petroleum ether/ethyl acetate=5:1, rf =0.4)) to give compound 12 (100 mg,254umol,22.9% yield) as a yellow solid, the structure was confirmed by1 H NMR.
LCMS product rt=1.229 mins, M/z= 393.2 (m+h)+
1H NMR(400MHz,CDCl3)δ9.25(s,1H),7.55-7.60(m,1H),7.49-7.51(m,1H),7.34-7.37(m,1H),4.53-4.57(m,1H),3.90-3.92(m,1H),2.15-2.20(m,2H),1.86-1.89(m,4H),1.60-1.64(m,2H),0.93(s,9H),0.08(s,6H).
2.3 Synthesis of Compound 12a
To a solution of compound 12-1 (2.00 g,14.2mmol,1.50mL,1.00 eq) and compound 12-1a (1.59 g,15.6mmol,2.03mL,1.10 eq) in dimethyl sulfoxide (10.0 mL) was added K2CO3 (3.92 g,28.4mmol,2.00 eq) and the resulting mixture was stirred at 40℃for 2 hours. TLC (petroleum ether: ethyl acetate=5:1) showed that compound 12-1 (Rf =0.6) remained and new product formation was detected. After the reaction was completed, the reaction mixture was diluted with H2 O (100 mL) and extracted with ethyl acetate (100 mL x 2), the combined organic layers were washed with saturated brine (200 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give compound 12-2 (3.20 g, crude oil), the structure was confirmed by1 H NMR for the next step without further purification.
1H NMR(400MHz,CDCl3)δ8.09-8.12(m,2H),6.59-6.62(m,2H),3.55(t,J＝7.2Hz,2H),3.10(s,1H),2.50(t,J＝7.2Hz,2H),2.30(s,6H).
To a solution of compound 12-2 (3.20 g,14.3mmol,1.00 eq.) in MeOH (30.0 mL) was added Pd/C (1.00 g,10% pure, 1.00 eq.). The suspension was degassed under vacuum and purged several times with hydrogen, after which the resulting solution was stirred under an atmosphere of H2 (15 psi) at 25 ℃ for 2 hours. TLC (dichloromethane: methanol=10:1) showed the starting point of compound 12-2 (rf=0.5) disappeared and new compound formation was detected. After the reaction was completed, the mixture was filtered and concentrated to give compound 12a (2.30 g,11.9mmol,83.0% yield) as a red-brown oil whose structure was confirmed by1 H NMR without further purification.
1H NMR(400MHz,DMSO)δ6.47-6.54(m,4H),4.34(br.s,2H),3.19(t,J＝7.2Hz,2H),2.72(s,1H),2.30(t,J＝7.2Hz,2H),2.14(s,6H).
2.4 Synthesis of Compound 13
To compound 12 (100 mg,254 mol,1.00 eq) and compound 12a (59.0 mg,305 mol,1.20 eq) IPA (3.00 mL) solution TFA (29.0 mg, 255. Mu. Mol, 18.8. Mu.L, 1.00 eq). The resulting mixture was stirred at 100 ℃ for 1 hour. LCMS showed the detection of the target product molecular weight (rt=0.913 min). After the reaction was completed, extracted with ethyl acetate (100 mL), the organic phase was washed with saturated NaHCO3 solution (50.0 mL), brine (50.0 mL), dried over anhydrous sodium sulfate, filtered and concentrated to give compound 13 (130 mg, crude), a yellow solid, which was directly used in the next step without purification.
2.5 Synthesis of Compound DD02013H
To a solution of compound 13 (130 mg,236umol,1.00 eq.) in DCM (5.00 mL) was added TFA (0.50 mL) and the resulting mixture was stirred at 25℃for 1 hour. LCMS showed the detection of the target product molecular weight (rt=0.701 min). After the reaction was completed, the mixture was concentrated to give a residual solid, which was purified by preparative high performance liquid chromatography, and the obtained solid was adjusted to ph=8 with saturated NaHCO3 solution, extracted with dcm (50.0 mL x 2), washed with brine (50.0 mL), dried over anhydrous sodium sulfate, filtered and concentrated to give yellow solid, the title compound DD02013H (47.6 mg,130.7umol,55.3% yield, 94.9% purity), the structure was confirmed by LCMS, HPLC and1 H-NMR.
LCMS product rt=0.997 min, M/z= 436.3 (m+h)+
HPLC product rt=2.951 mins, purity:95.6%
1H NMR(400MHz,CDCl3)δ9.00(s,1H),7.83-7.85(m,2H),7.29-7.31(m,1H),7.17-7.19(m,3H),6.85-6.87(m,2H),4.78-4.79(m,1H),3.76-3.81(m,1H),3.49(t,J＝7.2Hz,2H),2.95(s,3H),2.54(t,J＝7.2Hz,2H),2.33(s,6H),2.16-2.21(m,2H),2.03-2.06(m,2H),1.80-1.83(m,2H),1.69-1.73(m,2H).
EXAMPLE 3 Synthesis of the Compound DD02014H 4- ((2- ((1- (1-methylpiperidin-4-yl) -1H-pyrazol-4-yl) amino) quinazolin-8-yl) oxo) cyclohexan-1-ol
FIG. 3 shows the synthetic route for compound DD 02014H.
To a solution of compound 8 (400 mg,1.23mmol,1.00 eq) and compound 11a (569.1 mg,1.48mmol,1.20 eq) in DMF (10.0 mL) was added Cs2CO3 (1.00 g,3.08mmol,2.50 eq) and the resulting mixture was stirred at 80℃for 1 hour. LCMS showed the detection of the target product molecular weight (rt=0.958 min). After the reaction was completed, ethyl acetate (100 mL) was added, and the organic phase was washed with saturated NaHCO3 solution (50.0 mL), brine (50.0 mL), dried over anhydrous sodium sulfate, filtered and concentrated to give a residue, which was dispersed with petroleum ether: ethyl acetate (3:1, 20.0 mL) with shaking, filtered and dried to give yellow solid, compound 14 (300 mg,559umol,45.3% yield), the structure was characterized by LCMS.
LCMS product rt=0.912 min, M/z= 537.3 (m+h)+.
A mixture of compound 14 (300 mg,559umol,1.00 eq) and HCl/MeOH (4M, 10.0mL,71.6 eq) was stirred at 25℃for 1 hour, LCMS (EW 20001-117-P1A 1) showed the detection of the disappearance of the product, the appearance of the molecular weight of the target product (RT=0.729 min). After the reaction was completed, the mixture was concentrated, the resulting solid was purified by preparative HPLC, the mixture was then adjusted to ph=8 with saturated NaHCO3 solution, the dcm (50.0 mL x 2) was extracted, the organic phase was washed with brine (50.0 mL), dried over anhydrous sodium sulfate, filtered and concentrated to give yellow solid, compound DD02014H (35.5 mg,138umol,24.6% yield, 96.9% purity), the structure was confirmed by LCMS, HPLC and1 H NMR.
LCMS product rt=0.730 min, M/z= 423.2 (m+h)+
HPLC product rt=1.192mins, purity:98.0%
1H NMR(400MHz,CDCl3)δ9.01(s,1H),8.61(s,1H),7.49(s,1H),7.28-7.30(m,1H),7.17-7.22(m,3H),4.66-4.68(m,1H),4.23-4.30(m,1H),3.85-3.89(m,1H),2.99-3.03(m,2H),2.33(s,3H),2.25-2.29(m,4H),2.15-2.17(m,4H),1.95-2.06(m,2H),1.84-1.88(m,4H).
EXAMPLE 4 Synthesis of Compound DD02006H:4- ((2- ((4- (4-methylpiperazin-1-yl) phenyl) amino) quinazolin-8-yl) oxy) cyclohexan-1-ol
FIG. 4 shows the synthetic route for compound DD 02006H.
4.1 Synthesis of Compound 16
To a solution of compound 5 (1.00 g,5.14mmol,1.00 eq) and compound 16a (1.08 g,5.65mmol,1.10 eq) in IPA (20.0 mL) at 25 ℃ was added TFA (586 mg,5.14mmol,380ul,1.00 eq) and after mixing well, the reaction mixture was heated to 100 ℃ and stirred for 3 hours.
LCMS showed the molecular weight of the target product detected (rt=0.691 min). After the reaction was completed, the mixture was poured into 30.0mL of H2 O, then extracted with ethyl acetate (60.0 mL of x 4), the organic layer was washed with brine (60.0 mL of x 2), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to give compound 16 (1.50 g, crude) as a yellow solid. The product was used directly for the next step without purification.
LCMS product rt=0.691 min, M/z=350.2 (m+h)+
4.2 Synthesis of Compound 17
BBr3 (2.80 g,11.2mmol,1.08mL,3.00 eq) dissolved in DCM (10.0 mL) was added dropwise to a solution of compound 16 (1.30 g,3.72mmol,1.00 eq) in DCM (20.0 mL) at 0℃and after the addition was completed, stirred at 0℃for 30min, after which the mixture was heated to 25℃and stirred for 12h. LCMS showed the molecular weight of the target product detected (rt=0.677 min). After the reaction was completed, the mixture was poured into ice water (30.0 mL), extracted with ethyl acetate (60.0 mL x 3), the organic layer was washed with brine (30.0 mL x 2), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to give compound 17 (1.20 g, crude) as a grey solid. The product was used directly for the next step without purification.
LCMS product rt=0.677min, M/z=336.2 (m+h)+
4.3 Synthesis of Compound 18
Compound 17 (150 mg, 447. Mu. Mol,1.00 eq), compound 11a (276 mg, 894. Mu. Mol,2.00 eq) and Cs2CO3 (284 mg,1.12mmol,2.50 eq) were dissolved in DMF (10.0 mL) and the mixture was subsequently heated to 80℃for 12 hours. LCMS showed the detection of the target product molecular weight (rt=0.934 min). After the reaction was completed, the mixture was poured into 30.0mL of H2 O, then extracted with ethyl acetate (30.0 mL of x 4), and the organic layer brine (30.0 mL of x 2) was washed, dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to give compound 18 (200 mg, crude) as a yellow solid. The product was used directly for the next step without purification.
LCMS product rt=0.934min, M/z= 548.4 (m+h)+
4.4 Synthesis of Compound DD02006H
To a solution of compound 18 (200 mg,365umol,1.00 eq) in DCM (6.00 mL) was added dropwise HCl/1, 4-dioxane (4 m,2.00mL,21.9 eq) at 25 ℃ and the resulting mixture was stirred for 2 hours. LCMS showed the detection of the target product molecular weight (rt=0.746 min). Thin layer chromatography (DCM: meoh=8:1, material Rf =0.5, product Rf =0.2) showed new spot formation with no residual material. The mixture was poured into 30.0mL H2 O, pH adjusted to about 8 with saturated NaHCO3, then extracted with ethyl acetate (30.0 mL x 4), the organic layer washed with brine (30.0 mL x 2), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The crude product was concentrated under reduced pressure by pre-high performance liquid chromatography to give the desired product (68.6 mg,157umol,43.1% yield, 99.4% purity) as a yellow solid, the structure was confirmed by1 H NMR, LCMS and HPLC.
LCMS product rt=0.734min, M/z=434.3 (m+h)+;
HPLC product rt=1.373 mins,99.4% purity;
1H NMR(400MHz,DMSO)δ9.57-9.71(m,1H),9.12-9.27(m,1H),7.90-8.06(m,2H),7.40-7.47(m,1H),7.28-7.34(m,1H),7.18-7.26(m,1H),6.86-6.99(m,2H),4.75(br s,1H),4.52-4.64(m,1H),3.63(br s,1H),3.07(br d,J＝4.0Hz,4H),2.24(s,3H),1.91-2.05(m,2H),1.74-1.87(m,2H),1.56-1.72(m,4H).
EXAMPLE 5 Synthesis of the Compound DD02008H 8- (2-methoxyethoxy) -N- (4- (4-methylpiperazin-1-yl) phenyl) quinazolin-2-amine
FIG. 5 shows the synthetic route for compound DD 02008H.
Compound 17 (150 mg,447umol,1.00 eq), compound 17a (68.4 mg,492umol,46.2ul,1.10 eq) and Cs2CO3 (437mg, 1.34mmol,3.00 eq) were dissolved in DMF (10.0 mL), and the mixture was then heated to 80 ℃ and stirred for 12 hours, LCMS showed the detection of the target product molecular weight (rt=0.703 min). Thin layer chromatography (DCM: meoh=10:1, material rf=0.1, product rf=0.3) showed new spot formation with no starting material. After the reaction was completed, the mixture was poured into 30.0mL of H2 O, then extracted with ethyl acetate (30.0 mL of x 4), the organic layer was washed with brine (30.0 mL of x 2), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. Purification by column chromatography (SiO2, DCM/meoh=20/1 to 5/1) afforded the title compound DD02008H (71.94 mg,183umol,40.9% yield, 98.6% purity) as a yellow solid, structure confirmed by1 H NMR, LCMS and HPLC.
LCMS product rt=0.710 min, M/z=416.2 (m+h+na)+
HPLC product, rt=1.267 min,98.7% purity
1H NMR(400MHz,CDCl3)δ9.04(s,1H),7.72(br d,J＝8.0Hz,2H),7.33(dd,J＝8.0,4.0Hz,1H),7.15-7.24(m,2H),6.97(d,J＝8.0Hz,2H),4.33-4.41(m,2H),3.91-3.99(m,2H),3.56(s,3H),3.13-3.25(m,4H),2.58-2.65(m,4H),2.38(s,3H).
EXAMPLE 6 Synthesis of Compound DD02015H 3- ((2- ((4- (4-methylpiperazin-1-yl) phenyl) amino) quinazolin-8-yl) oxo) cyclopentan-1-ol
FIG. 6 shows the synthetic route for compound DD 02015H.
Compound 17 (150 mg,447umol,1.00 eq), compound 18a (331 mg,894umol,2.00 eq) and Cs2CO3 (284 mg,1.12mmol,2.50 eq) were dissolved in DMF (10.0 mL) and the mixture was then heated to 80 ℃ and reacted with stirring for 12 hours. LCMS showed the detection of the target product molecular weight (rt=0.893 min), TLC (DCM: meoh=10:1, material rf=0.1, product rf=0.25) showed new spot formation, no starting material. After the reaction was completed, the resulting mixture was poured into 30mL of water, followed by extraction with ethyl acetate (30.0 mL of 4), washing with brine (30.0 mL of 2), drying over anhydrous sodium sulfate, filtration, removal of the solvent under reduced pressure, and purification of the resulting residual solid by column chromatography (SiO 2, DCM/meoh=20/1 to 5/1) gave compound 19 (200 mg,375umol,83.8% yield) as a yellow solid. LCMS product rt=0.893 min, M/z= 534.4 (m+h)+
Compound 19 (200 mg,375umol,1.00 eq) was dissolved in DCM (6.00 mL) and HCl/1, 4-dioxane (4M, 4.00mL,42.7 eq) was added dropwise at 25℃and the reaction was stirred for 2 hours. LCMS showed the detection of the target product molecular weight (rt=0.721 min) and hplc showed the starting material reaction was complete. After the reaction was completed, the mixture was poured into 30.0mL H2 O, adjusted to pH about 8 with saturated NaHCO3, then extracted with ethyl acetate (30.0 mL x 4), the organic layer brine (30.0 mL x 2) was washed, dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure, and the resulting crude product was purified by preparative high performance liquid chromatography to give the title compound DD02015H (110 mg,262umol,70.0% yield, 100% purity) as a yellow solid, the structure was confirmed by1 H NMR, LCMS and HPLC.
LCMS product rt=0.716min, M/z=420.3 (m+h)+
HPLC product rt=1.284 min,100% purity.
1H NMR(400MHz,CDCl3)δ9.04(s,1H),7.60(br d,J＝8.0Hz,2H),7.32-7.38(m,1H),7.21-7.24(m,2H),6.93-7.01(m,2H),5.15(t,J＝4.0Hz,1H),4.41(br t,J＝4.0Hz,1H),3.17-3.26(m,4H),2.62(m,4H),2.38(s,3H),1.94-2.20(m,6H).
EXAMPLE 7 Synthesis of the Compound DD02021H 4- ((2- ((5- (4-methylpiperazin-1-yl) pyridin-2-yl) amino) quinazolin-8-yl) oxo) cyclohexan-1-ol
FIG. 7 shows the synthetic route for compound DD 02021H.
7.1 Synthesis of Compound 20
Compound 5 (500 mg,2.57 mmol,1.00 eq), compound 19a (988 mg,5.14 mmol,2.00 eq), pd2(dba)3 (353 mg,386 umol,0.15 eq) and BINAP (184 mg,295 umol,1.15e-1 eq) were dissolved in 1, 4-dioxane (30.0 mL) at 25℃and Cs2CO3 (1.67 g,5.14 mmol,2.00 eq) was added to the resulting mixture, and the reaction mixture was then heated to 100 ℃
And stirred for 10 hours. LCMS showed the detection of the target product molecular weight (rt=0.643 min). After the reaction was completed, the mixture was poured into 30.0 mL H2 O, then extracted with DCM (50.0 ml×3), the organic layer brine (30.0 ml×3) was washed, dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to give compound 20 (600 mg, crude) as a yellow solid. The product was used directly in the next reaction without purification.
7.2 Synthesis of Compound 21
Compound 20 (600 mg,1.71mmol,1.00 eq) was dissolved in DCM (10.0 mL) and the resulting solution was added dropwise BBr3 (428 mg,1.71mmol,165uL,1.00 eq) dissolved in DCM (5.00 mL) and the system was stirred at 25℃for 12 hours after the addition. LCMS showed the detection of the target product molecular weight (rt=0.321 min). After the reaction was completed, the mixture was poured into ice water (40.0 mL), pH was adjusted to about 8 with saturated NaHCO3, then extracted with DCM (50.0 mL x 3), the organic layer was washed with brine (30.0 mL x 3), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give compound 21 (600 mg, crude) as a yellow solid. The product was used directly in the next reaction without purification. LCMS product rt=0.324 min, M/z= 337.2 (m+h)+
7.3 Synthesis of Compound 22
Compound 21 (600 mg,1.78mmol,1.00 eq), compound 11a (1.37 g,3.57mmol,2.00 eq) and Cs2CO3 (1.45 g,4.46mmol,2.50 eq) were dissolved in DMF (10.0 mL) and the resulting mixture was heated to 80℃for 12 hours. LCMS showed the detection of the target product molecular weight (rt=0.830 min), thin layer chromatography (DCM: meoh=10:1, material Rf =0.1, product Rf =0.3) showed new spot formation with no starting material. After the reaction was completed, the mixture was poured into 30.0mL of H2 O, then extracted with ethyl acetate (60.0 mL of x 3), the organic layer brine (40.0 mL of x 2), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure, and the resulting material was purified by column chromatography (SiO2, DCM/meoh=40/1 to 5/1) to give compound 22 (700 mg,1.08mmol,60.5% yield, 84.6% purity) as a yellow solid.
LCMS product rt=0.830 min, M/z= 549.4 (m+h)+
HPLC product rt=2.563 mins,84.6% purity.
7.4 Synthesis of Compound DD02021H
Compound 22 (600 mg,925umol,1.00 eq.) and HCl/1, 4-dioxane (4M, 6.00mL,25.9 eq.) were dissolved in DCM (10.0 mL) and the resulting mixture was stirred at 25℃for 1 hour. LCMS showed the detection of the target product molecular weight (rt=0.680 min), thin layer chromatography (DCM: meoh=10:1, material Rf =0.3, product Rf =0.15) showed new spot formation, no starting material. After the reaction was completed, the mixture was slowly poured into 40.0mL of H2 O with stirring, then extracted with ethyl acetate (80.0 mL x 3), the organic layer brine (50.0 mL x 2) was washed, dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The resulting solid was purified by preparative HPLC, the pH of the material was adjusted to about 8 with saturated NaHCO3, then extracted with DCM (40.0 ml x 4), the organic layer was washed with brine (40.0 ml x 3), dried over anhydrous Na2SO4, filtered under reduced pressure and concentrated to give the target product DD02021H, (148 mg,329umol,35.6% yield, 96.7% purity) as a yellow solid, the structure confirmed by1 H NMR, LCMS and HPLC.
LCMS product rt=0.7197min, M/z= 435.3 (m+h)+
HPLC product rt=1.232 min,96.7% purity
1H NMR(400MHz,DMSO)δ9.74(s,1H),9.26(s,1H),8.86(d,J＝8.0Hz,1H),8.02(d,J＝4.0Hz,1H),7.43-7.57(m,2H),7.24-7.40(m,2H),4.78(br s,1H),4.62-4.73(m,1H),3.54-3.69(m,1H),3.08-3.16(m,4H),2.47(m,3H),2.23(s,3H),1.93-2.04(m,2H),1.74-1.88(m,2H),1.58-1.72(m,4H).
EXAMPLE 8 Synthesis of the Compound DD02018H 4- ((8- ((4-hydroxycyclohexyl) oxo) quinazolin-2-yl) amino) -N- (1-methylpiperidin-4-yl) benzamide
FIG. 8 shows the synthetic route for compound DD 02018H.
Compound 11 (400 mg,2.21mmol,1.00 eq), compound 11a (1.28 g,3.32mmol,1.50 eq) and Cs2CO3 (1.44 g,4.43mmol,2.00 eq) were dissolved in DMF (10.0 mL) and the resulting mixture was heated to 80℃and stirred for 2 hours. LCMS showed the detection of the target product molecular weight (rt=1.218 min). After the reaction was completed, the mixture was slowly poured into 40.0mL of H2 O, then extracted with DCM (80.0 mL of x 3), the organic layer brine (50.0 mL of x 2) was washed, dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to give compound 15 (310 mg, crude) as a grey solid. The product was used directly in the next reaction without purification.
Compound 15 (260 mg,661 mol,1.00 eq), compound 15a (231.53 mg,992.38 mol,1.50 eq) and TFA (75.4 mg,661 mol,49.0ul,1.00 eq) were dissolved in IPA (10.0 mL), and the resulting mixture was then heated to 80 ℃ and stirred for 2 hours, and the mixture was heated to 100 ℃ and stirred for 2 hours. LCMS showed the detection of the target product molecular weight (rt=0.762 min) and high performance liquid chromatography showed a purity of 59.7% (rt=1.535 min). After the reaction was completed, the mixture was slowly poured into 40.0mL of H2 O with stirring, pH was adjusted to about 8 with saturated NaHCO3, then extracted with DCM (80.0 mL of x 3), the organic layer was washed with brine (50.0 mL of x 2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure, the resulting crude product was purified by preparative high performance liquid chromatography, the resulting material was adjusted to pH 8 with saturated NaHCO3, extracted with DCM (40.0 mL of x 4), the organic layer was washed with brine (40.0 mL of x 3), dried over anhydrous Na2SO4, filtered and concentrated to give the target product DD02018H (17.1 mg,35.13umol,5.31% yield, 97.7% purity) as a pale yellow solid, the structure was confirmed by1 H NMR, s and HPLC.
LCMS product rt=0.771 min, M/z=476.2 (m+h)+
HPLC product rt=1.53mins, 97.3% purity
1H NMR(400MHz,DMSO-d6)δ10.15(br s,1H),9.30(br s,1H),8.23(br d,J＝8.0Hz,2H),8.13(br s,1H),7.83(br d,J＝8.0Hz,2H),7.49(br s,1H),7.27-7.42(m,2H),4.73(br s,1H),4.60(br s,1H),3.87(br s,1H),3.07(br s,2H),2.62-2.67(m,2H),2.33(br s,3H),1.84-2.08(m,3H),1.73(m,9H).
EXAMPLE 9 Synthesis of the Compound DD02002H 4- ((8- ((3-hydroxycyclopentyl) oxo) quinazolin-2-yl) amino) benzenesulfonamide
FIG. 9 shows the synthetic route for compound DD 02002H.
9.1 Synthesis of Compound 23
Compound 11 (200 mg,1.11 mmol) was dissolved in anhydrous tetrahydrofuran (5 mL) under nitrogen, triphenylphosphine (581 mg,2.22 mmol), DIAD (449 mg,2.22 mmol) was added sequentially, stirring was carried out for 15min, and finally 1, 3-cyclopentanediol (i.e. compound 22,340.1mg,3.33 mmol) was added, the reaction was stirred overnight at room temperature, and TLC (PE/EA=3:1) analysis showed complete consumption of starting material with a new product. After the completion of the reaction, the resulting mixture was concentrated to remove the solvent, and the crude product was separated by a silica gel column to give compound 23 (210 mg), yield 71.5%, as a pale yellow solid.
LCMS product rt=2.76 min, M/z= 265.1 (m+h)+
9.2 Synthesis of Compound DD02002H
Compound 23 (200 mg,0.76 mmol) was dissolved in isopropanol (5 mL) under nitrogen, sulfanilide (260.2 mg,1.51 mmol) was added, the reaction was heated to 90℃overnight, thin layer chromatography analysis showed some starting material remained, new product spots were formed, and LCMS showed the detection of the molecular weight of the target product. After the reaction, the crude product was concentrated, and the target molecule DD02002H (104 mg) was obtained by separation on a silica gel column in 34% yield as a white solid whose structure was confirmed by1 H NMR and LCMS.
LCMS product rt=3.46 min, M/z=401.1 (m+h)+
1H NMR(400MHz,MeOD)δ9.21(s,1H),8.23-8.25(m,2H),7.87-7.89(m,2H),7.48(d,J＝7.6Hz,1H),7.33-7.37(m,2H),4.36(m,1H),4.20(m,1H),1.76-2.10(m,6H).
EXAMPLE 10 Synthesis of the Compound DD02019H:3- ((8- ((4-hydroxycyclohexyl) oxo) quinazolin-2-yl) amino) benzenesulfonamide
FIG. 10 shows the synthetic route for compound DD 02019H.
10.1 Synthesis of Compound 26
Compound 11 (200 mg,1.11 mmol) was dissolved in anhydrous tetrahydrofuran (5 mL), triphenylphosphine (581 mg,2.22 mmol) was added sequentially, DIAD (449 mg,2.22 mmol) was stirred for 15min, 1, 4-cyclohexanediol (387 mg,3.33 mmol) was added finally, the resulting reaction mixture was stirred overnight at room temperature, TLC (PE/EA=3:1) analysis showed complete consumption of starting material with new product spot formation, LCMS detected the target product molecular weight (RT=4.03 min), the reaction mixture was concentrated, and the resulting crude product was isolated as compound 26 (197 mg) via a silica gel column in 70.7% yield as a pale yellow solid. LCMS product rt=4.03min, M/z=279.1 (m+h)+
10.2 Synthesis of Compound DD02019H
Compound 26 (150 mg,0.54 mmol) was dissolved in isopropanol (5 mL) under nitrogen, meta-aminobenzenesulfonamide (185.4 mg,1.1 mmol) was added, the reaction was heated to 90℃overnight, TLC analysis showed the starting material remained with a new product spot, LC-MS detected the molecular weight of the target product, the reaction solution was concentrated, the crude product obtained was separated by a silica gel column to give the target molecule DD02019H (27 mg), yield 12.1% was a white solid, the structure was confirmed by LC-MS and1 H NMR. LCMS product rt=3.54 min, M/z=415.2 (m+h)+
1H NMR(400MHz,MeOD)δ9.19(s,1H),8.51(m,2H),7.34-7.55(m,5H),4.62(m,1H),3.73(m,1H),1.57-1.94(m,8H).
Example 11 in vitro test of Compounds for inhibiting the Activity of HPK1 kinase
Table 1 shows the reagents and sources thereof required for the experiment
All compounds were tested at an initial concentration of 10 μm, diluted in a three-fold concentration gradient, for a total of 10 concentration points, each concentration being repeated once.
11.1 Compounds in vitro inhibition of HPK1 kinase Activity assay
Before the start of the experiment, 50. Mu.M DTT and enzymatic reaction system buffer were prepared.
Transferring the compound diluent into a 384-well plate by a pipette, sealing the 384-well plate after adding, centrifuging for 1min at the speed of 1000g, preparing 2-fold concentration HPK1 solution in a kinase buffer, adding 2.5 mu L of prepared 2-fold concentration HPK1 solution into the 384-well plate, centrifuging for 30s at the speed of 1000g, incubating for 10min at room temperature, preparing 2-fold concentration MBP and ATP mixed solution in the kinase buffer, adding 2.5 mu L of prepared 2-fold concentration MBP and ATP mixed solution into the reaction system, centrifuging for 30s at the speed of 1000g, incubating for 1h at room temperature, adding 5 mu L of ADP-Glo reagent into the reaction system, incubating for 40min at room temperature, adding 10 mu L of kinase detection reagent, incubating for 40min at room temperature, and reading luminescence signals on an Envision 2104plate reader, and calculating the inhibition rate according to the following formula:
%inhibition=100-(Signalcmpd-SignalAve_PC)/(SignalAve_VC-SignalAve_PC)×100
In the above formula, cmpd refers to the test compound, PC refers to the positive control, and VC refers to the negative control. The positive control used for the HPK1 experiment was Sunitinib.
Table 2 shows the inhibition of HPK1 kinase activity by the compounds described in the examples