The invention still further relates to the purposes of this Emulsion in the medicine of preparation treatment or prevent diabetes or blood sugar lowering.The invention still further relates to the method for a kind of treatment or prevent diabetes or blood sugar lowering, comprise the step of this Emulsion that gives effective dose.In a specific embodiment of the present invention, the component and the content of the described Emulsion that contains the hydrophilic biomacromolecule are as follows:

Recombinant human insulin's 0.04 weight portion

Sodium hydroxide is an amount of

Deionized water 1 weight portion

Dl-alpha-tocopherol acetate 1 weight portion

Medium chain length fatty acid triglyceride (Labrafac 0.5 weight portion Lipophile 1349 for example, Gattefosse)

Polyethylene Glycol caprylic/capric glyceride (3 weight portions for example

767, SASOL)

Poloxamer 188 0.2 weight portions

Soybean lecithin 0.5 weight portion.

The invention still further relates to the purposes of this Emulsion in the medicine of preparation treatment or prevent diabetes or blood sugar lowering.

GLP-1 0.0005 weight portion

PBS buffer 0.3 weight portion

D-alpha-tocopherol succinic acid macrogol ester 1.5 weight portions

Polyethyleneglycol-12-hydroxy stearin 2 weight portions

Polysorbas20 0.5 weight portion.

The invention still further relates to the purposes of this Emulsion in the medicine of preparation treatment or prevent diabetes or blood sugar lowering.The invention still further relates to the method for a kind of treatment or prevent diabetes or blood sugar lowering, comprise the step of this Emulsion that gives effective dose.

PTH 1-34 0.005-0.015 weight portion

Water (preferred distilled water) 0.4-0.6 weight portion

Acetic acid is an amount of

Dl-alpha-tocopherol 0.6-0.8 weight portion

D-alpha-tocopherol succinic acid macrogol ester 0.4-0.6 weight portion

Soybean oil 1.0-1.4 weight portion

Soybean lecithin 0.6-0.8 weight portion

Polyoxyethylene ether (40) castor oil hydrogenated 2.5-3.5 weight portion

PEG400 1.5-2.5 weight portion.

PTH 1-34 0.01 weight portion

Water (preferred distilled water) 0.5 weight portion

Acetic acid is an amount of

Dl-alpha-tocopherol 0.7 weight portion

D-alpha-tocopherol succinic acid macrogol ester 0.5 weight portion

Soybean oil 1.2 weight portions

Soybean lecithin 0.7 weight portion

Polyoxyethylene ether (40) castor oil hydrogenated 3 weight portions

PEG400 2 weight portions.

The invention still further relates to the purposes of this Emulsion in the medicine of preparation treatment or prevention of osteoporosis.

Low molecular weight heparin 0.3 weight portion

PBS buffer 0.5 weight portion

Dl-alpha-tocopherol 0.2 weight portion

Polyglycereol-3-oleate 1.2 weight portions

Tween 80 1 weight portion

Polyethylene Glycol caprylic/capric glyceride 2.7 weight portions

PEG400 0.5 weight portion.

The invention still further relates to this Emulsion the preparation anticoagulation and or the medicine of thrombolytic in purposes.The invention still further relates to a kind of treatment or pre-preventing thrombosis or anticoagulant method, comprise the step of this Emulsion that gives effective dose.

Small molecules interference RNA (for example size is 500bp) 0.0003 weight portion

Water for injection 0.5 weight portion

Dl-alpha-tocopherol 0.2 weight portion

Polyglycereol-3-oleate 0.3 weight portion

Polyethylene Glycol caprylic/capric glyceride 2.7 weight portions

Glycerol 0.5 weight portion.

Another aspect of the present invention relates to each described preparation method that contains the Emulsion of hydrophilic biomacromolecule of the present invention, comprises the steps:

1) under 4-25 ℃ of condition, the hydrophilic biomacromolecule is soluble in the aqueous phase;

2) under 20-70 ℃ of condition, esters derivative and oil phase, the emulsifying agent of vitamin E and/or vitamin E mixed mutually, and under the 200-800rmp stirring condition, continue to stir, be the transparent clear shape until whole system;

3) under 4-37 ℃ of condition,, dropwise add step 2 with the step 1) product) in the product, and under the 200-800rmp condition, continue to stir, until the even preparation that forms transparent clear, the final Emulsion that obtains to contain the hydrophilic biomacromolecule.

The purposes of the Emulsion that contains the hydrophilic biomacromolecule of the present invention in the medicine of preparation treatment or prevent diabetes or blood sugar lowering that also relate in one aspect to of the present invention, wherein, described hydrophilic biomacromolecule is insulin or GLP-1.

The purposes of the Emulsion that contains the hydrophilic biomacromolecule of the present invention in the medicine of preparation treatment or prevention of osteoporosis that also relate in one aspect to of the present invention, wherein, described hydrophilic biomacromolecule is calcitonin, parathyroid hormone, parathyroid hormone 1-84 or human parathyroid hormone 1-34.

The esters derivative that also relates in one aspect to vitamin E and/or vitamin E of the present invention contains purposes in the Emulsion of hydrophilic biomacromolecule in preparation.Particularly, the described Emulsion that contains the hydrophilic biomacromolecule is each described Emulsion that contains the hydrophilic biomacromolecule of the present invention.Particularly, described vitamin E is to be selected from d-alpha-tocopherol and the dl-alpha-tocopherol one or more, and the esters derivative of described vitamin E is to be selected from d-alpha-tocopherol acetate, dl-alpha-tocopherol acetate, d-alpha-tocopherol succinate, dl-alpha-tocopherol succinate, d-alpha-tocopherol succinic acid macrogol ester and the dl-alpha-tocopherol succinic acid macrogol ester one or more.Particularly, described hydrophilic biomacromolecule and water, oil phase, emulsifying agent, pH regulator agent etc. and consumption thereof as mentioned described in.

The beneficial effect of the invention

The present invention compared with prior art has the following advantages and the salience effect:

1) among the present invention, esters derivative by means of vitamin E and/or vitamin E, realize the oralization administration of hydrophilic biopharmaceutical macromolecular drug, and have the characteristics safe and effective, that oral administration biaavailability is high, bioavailability can reach more than 5%, 10%, 12%, 15% even 30%;

2) the vitamin E microemulsion that contains the biological low-molecule drug of hydrophilic that makes can spontaneous emulsification be the following emulsion droplet of 300nm behind the chance water, is better utilized by gastrointestinal absorption, brings into play better biological activity.

3) preparation technology of the present invention is simple, and is with low cost, is suitable for large-scale industrial production.

Description of drawings

Fig. 1: under the different modes of administration, the rat blood sugar variation diagram.

Fig. 2: under the different modes of administration, phasor during P of Rats TH1-34 concentration.

The specific embodiment

Below in conjunction with embodiment embodiment of the present invention are described in detail, but it will be understood to those of skill in the art that the following example only is used to illustrate the present invention, and should not be considered as limiting scope of the present invention.Unreceipted actual conditions person among the embodiment carries out according to the condition of normal condition or manufacturer's suggestion.The unreceipted person of production firm of agents useful for same or instrument, being can be by the conventional products of commercial acquisition.

Embodiment 1: contain the preparation of the Emulsion (microemulsion) of salmon calcitonin see calcimar

It is composed as follows:

Salmon calcitonin see calcimar 0.001 gram

1 mole hydrochloride is an amount of

Normal saline 0.35 gram

D-alpha-tocopherol 0.7 gram

Oleic acid 0.3 gram

Polyethyleneglycol-12-hydroxy stearin (Solutol 1.5gram HS 15, BASF)

Tween 80 1.5 grams.

The preparation method of said preparation is summarized as follows:

1) under 4 ℃, salmon calcitonin see calcimar is dissolved in normal saline, adds appropriate hydrochloric acid, stirs to clarify, A solution;

2) under 70 ℃,, mix being incorporated under the 200rpm stirring condition, be stirred to transparent clear d-alpha-tocopherol, oleic acid, polyethyleneglycol-12-hydroxy stearin, the Tween 80 of above-mentioned weight;

3) under 30 ℃,, dropwise join step 2 with A solution) product in, under the 800r/min rotating speed, mixture is stirred to transparent clear, the final Emulsion (microemulsion) (being called sample 1) that obtains to contain salmon calcitonin see calcimar.

Although it will be understood by those skilled in the art that the unit of weight of each component among the embodiment 1 is g, also can be understood as weight portion, the part by weight among the embodiment 1 above promptly satisfying between each component just can.

Embodiment 2: contain the preparation of recombinant human insulin's Emulsion (microemulsion)

It is composed as follows:

Recombinant human insulin's 0.04 gram

5 molar sodium hydroxides are an amount of

Deionized water 1 gram

Dl-alpha-tocopherol acetate 1 gram

Medium chain length fatty acid triglyceride (Labrafac Lipophile 0.5 gram 1349, Gattefosse)

Polyethylene Glycol caprylic/capric glyceride (

3 grams 767, SASOL)

Poloxamer 188 (Lutrol F68, BASF) 0.2 gram

(Epikuron 170, Degussa) 0.5 gram for soybean lecithin.

The preparation method of said preparation is summarized as follows:

1) under 25 ℃, the recombinant human insulin of above-mentioned weight is dissolved in 0.5 gram deionized water, adds an amount of sodium hydroxide, stir to clarify, A solution;

2) under 20 ℃, dl-alpha-tocopherol acetate, medium chain length fatty acid triglyceride, Polyethylene Glycol caprylic/capric glyceride, poloxamer 188, soybean lecithin with above-mentioned weight, mix with the residue water, and under the 800rpm stirring condition, be stirred to transparent clear;

3) under 4 ℃,, dropwise join step 2 with A solution) product in, under the 200r/min rotating speed, mixture is stirred to transparent clear, the final Emulsion (microemulsion) (being called sample 2) that obtains to contain the recombinant human insulin.

Although it will be understood by those skilled in the art that the unit of weight of each component among the embodiment 2 is g, also can be understood as weight portion, the part by weight among the embodiment 2 above promptly satisfying between each component just can.

Embodiment 3: contain the preparation of the Emulsion (microemulsion) of GLP-1

It is composed as follows:

GLP-1 0.0005 gram

PBS buffer 0.3 gram

D-alpha-tocopherol succinic acid macrogol ester (TPGS, 1.5 gram Eastman)

Polyethyleneglycol-12-hydroxy stearin (Solutol HS 2grams 15, BASF)

Polysorbas20 0.5 gram.

The preparation method of said preparation is summarized as follows:

1) under 20 ℃, the GLP-1 of above-mentioned weight is dissolved in the PBS buffer, stirs to clarify, A solution;

2) under 60 ℃,, mix to be incorporated under the 600rpm condition being stirred to transparent clear with d-alpha-tocopherol succinic acid macrogol ester, polyethyleneglycol-12-hydroxy stearin, the polysorbas20 of above-mentioned weight;

3) under 37 ℃,, dropwise join step 2 with A solution) product in, under the 500r/min condition, mixture is stirred to transparent clear, the final Emulsion (microemulsion) (being called sample 3) that obtains to contain GLP-1.

Although it will be understood by those skilled in the art that the unit of weight of each component among the embodiment 3 is g, also can be understood as weight portion, the part by weight among the embodiment 3 above promptly satisfying between each component just can.

Embodiment 4: contain the preparation of the Emulsion (microemulsion) of PTH 1-34

It is composed as follows:

PTH 1-34 0.01 gram

Distilled water 0.5 gram

Acetic acid is an amount of

Dl-alpha-tocopherol 0.7 gram

D-alpha-tocopherol succinic acid macrogol ester (TPGS, 0.5 gram Eastman)

Soybean oil 1.2 grams

(Epikuron 170, Degussa) 0.7 gram for soybean lecithin

Polyoxyethylene ether (40) castor oil hydrogenated (Cremophor 3 grams40, BASF)

PEG400 2 grams.

The preparation method of said preparation is summarized as follows:

1) under 20 ℃, the PTH 1-34 of above-mentioned weight is dissolved in the distilled water, adds an amount of acetic acid, liquid stirs to clarify, A solution;

2) under 50 ℃, with dl-alpha-tocopherol, d-alpha-tocopherol succinic acid macrogol ester, soybean oil, soybean lecithin, polyoxyethylene ether (40) castor oil hydrogenated, the PEG400 of above-mentioned weight, mix to be incorporated under the 400rpm condition being stirred to transparent clear;

3) under 30 ℃,, dropwise join step 2 with A solution) product in, and under the 500r/min rotating speed, continue to be stirred to transparent clear, the final Emulsion (microemulsion) (being called sample 4) that obtains to contain PTH 1-34.

Although it will be understood by those skilled in the art that the unit of weight of each component among the embodiment 4 is g, also can be understood as weight portion, the part by weight among the embodiment 4 above promptly satisfying between each component just can.

Embodiment 5: contain the preparation of the Emulsion (microemulsion) of low molecular weight heparin

It is composed as follows:

Low molecular weight heparin 0.3 gram

PBS buffer 0.5 gram

Dl-alpha-tocopherol 0.2 gram

Polyglycereol-3-oleate 1.2 grams

Tween 80 1 gram

Polyethylene Glycol caprylic/capric glyceride 2.7 grams

PEG400 0.5 gram.

The preparation method of said preparation is summarized as follows:

1) under 10 ℃, the low molecular weight heparin of above-mentioned weight is dissolved in PBS solution, stirs to clarify, A solution;

2) under 50 ℃,, mix to be incorporated under the 400rpm condition being stirred to transparent clear with dl-alpha-tocopherol, polyglycereol-3-oleate, Tween 80, Polyethylene Glycol caprylic/capric glyceride, the PEG400 of above-mentioned weight;

3) under 30 ℃,, dropwise join step 2 with A solution) product in, and under the 500r/min rotating speed, continue to be stirred to transparent clear, the final Emulsion (microemulsion) (being called sample 5) that obtains to contain low molecular weight heparin.

Although it will be understood by those skilled in the art that the unit of weight of each component among the embodiment 5 is g, also can be understood as weight portion, the part by weight among the embodiment 5 above promptly satisfying between each component just can.

Embodiment 6: contain the preparation of the Emulsion (microemulsion) of small molecules interference RNA

It is composed as follows:

(self-control derives from the malicious P of sick 0.0003 gram of hepatitis B district, 500bp) to small molecules interference RNA

Water for injection 0.5 gram

Dl-alpha-tocopherol 0.2 gram

Polyglycereol-3-oleate 0.3 gram

Polyethylene Glycol caprylic/capric glyceride 2.7 grams

Glycerol 0.5 gram.

The preparation method of said preparation is summarized as follows:

1) under 4 ℃, the small molecules interference RNA of above-mentioned weight is dissolved in water for injection, stirs to clarify, A solution;

2) under 50 ℃,, mix to be incorporated under the 300rpm condition being stirred to transparent clear with dl-alpha-tocopherol, polyglycereol-3-oleate, Polyethylene Glycol caprylic/capric glyceride, the glycerol of above-mentioned weight;

3) under 10 ℃,, dropwise join step 2 with A solution) product in, and under the 200r/min rotating speed, continue to be stirred to transparent clear, the final Emulsion (microemulsion) (being called sample 6) that obtains to contain small molecules interference RNA.

Although it will be understood by those skilled in the art that the unit of weight of each component among the embodiment 6 is g, also can be understood as weight portion, the part by weight among the embodiment 6 above promptly satisfying between each component just can.

Embodiment 7: the particle size determination test

Detect the particle diameter of the sample 1-6 of embodiment 1-6 preparation with Malvern Zetasizer 3000HSA Particle Size Analyzer, before the mensuration, sample is with 30 times of Millipore ultra-pure water dilutions, and the result is as shown in table 1.

Table 1: the mean diameter after the sample 1-6 dilution

Sample number into spectrum	Particle diameter/nm
		1	170
2	68
		3	267
4	235
		5	182
6	280

Embodiment 8: the test of pesticide effectiveness that contains recombinant human insulin's Emulsion (microemulsion)

6 of the male SD rats of oral test group: 180-220 gram scope, medicine (trial drug) are the prepared sample 2 of embodiment 2.

6 of the male SD rats of oral matched group: 180-220 gram scope, medicine (control drug) are pressed composition and the operating procedure preparation of embodiment 2, except not adding d1-alpha-tocopherol acetate.

6 of the male SD rats of injection matched group: 180-220 gram scope, medicine is an insulin solution.

Test method:

Be to begin test behind the male SD rat fasting 12h of 180-220 gram scope with body weight.Test group is an oral administration, and dosage is counted 0.4mg/kg by insulin.Rats in test groups is after the pentobarbital sodium of 50mg is anaesthetized rat, by facing upward a mode it to be bundled, and cut an osculum at abdominal part with dosage before the test,,, sews up the incision after the administration to the ileum administration with administrator.Matched group is the subcutaneous injection control drug, and dosage is counted 0.04mg/kg by insulin.

Three groups of groups all after administration 0.5,1,2,3, the 4h moment point, get blood by the tail vein, use Luo Shi blood sugar detection instrument to measure blood glucose and calculate every group in the average blood sugar values of 6 rats, the result is as by shown in Figure 1.As seen from Figure 1, the test group of oral administration is 12.5% with respect to the bioavailability of the matched group of drug administration by injection.

Embodiment 9: contain Emulsion (microemulsion) the bioavailability test of PTH1-34

10 of the male SD rats of test group: 180-220 gram scope, medicine (trial drug) are the prepared sample 4 of embodiment 4.

10 of the male SD rats of matched group: 180-220 gram scope, medicine is the PTH1-34 aqueous solution, takes drug administration by injection.

Test method:

Be to begin test behind the male SD rat fasting 12h of 180-220 gram scope with body weight.The test group oral administration, dosage is counted 100 μ g/kg by PTH1-34.Oral treated animal is after the pentobarbital sodium of 50mg is anaesthetized animal, by facing upward a mode it to be bundled, and cut an osculum at abdominal part with dosage before the experiment,,, sews up the incision after the administration to the ileum administration with administrator.Matched group is subcutaneous injection PTH1-34, and dosage is counted 10 μ g/kg by PTH1-34.

Two groups all after

administration

15,30,45,60,75,90, the moment point of 105min, get blood by the tail vein, isolate serum and be equipped with and survey.PTH1-34 concentration detects with PTH1-34 high sensitivity EIA test kit in the serum.The result as shown in Figure 2, areameter under the PTH1-34 plasma concentration curve wherein.As seen from Figure 2, the test group of oral administration is 23.13% with respect to the bioavailability of the matched group of drug administration by injection.

Although the specific embodiment of the present invention has obtained detailed description, it will be understood to those of skill in the art that.According to disclosed all instructions, can carry out various modifications and replacement to those details, these change all within protection scope of the present invention.Four corner of the present invention is provided by claims and any equivalent thereof.

Claims

1. an Emulsion that contains the hydrophilic biomacromolecule comprises hydrophilic biomacromolecule, water, oil phase, emulsifying agent, it is characterized in that, described Emulsion also comprises the esters derivative of vitamin E and/or vitamin E.

2. Emulsion according to claim 1, wherein, described hydrophilic biomacromolecule is selected from protein or polypeptide, polysaccharide and nucleic acid.

3. Emulsion according to claim 1, wherein, described vitamin E is to be selected from d-alpha-tocopherol and the dl-alpha-tocopherol one or more, and the esters derivative of described vitamin E is to be selected from d-alpha-tocopherol acetate, dl-alpha-tocopherol acetate, d-alpha-tocopherol succinate, dl-alpha-tocopherol succinate, d-alpha-tocopherol succinic acid macrogol ester and the dl-alpha-tocopherol succinic acid macrogol ester one or more.

4. Emulsion according to claim 1, wherein, described hydrophilic biomacromolecule is the 0.01-1 weight portion, and the esters derivative of described vitamin E and/or vitamin E is 50-1000 weight portion, 50-500 weight portion or 100-300 weight portion.

5. Emulsion according to claim 4, wherein, described water, oil phase and emulsifying agent add up to 20-2000 weight portion, 20-1000 weight portion or 50-500 weight portion.

6. Emulsion according to claim 1, wherein, described protein or polypeptide are selected from calcitonin, insulin, GLP-1, parathyroid hormone and growth hormone; Described polysaccharide is selected from heparin, Low molecular heparin, chondroitin sulfate, keratan sulfate and hyaluronic acid; Described nucleic acid is selected from antisensenucleic acids, ribozyme, RNA interfering, DECOY nucleic acid and three chain nucleic acid; Particularly; Described calcitonin is salmon calcitonin see calcimar, Anguillar japonica calcitonin or human calcitonin; Described insulin is Iletin II (Lilly), bovine insulin or insulin human; Described parathyroid hormone is that human body parathyroid hormone 1-84 or human parathyroid hormone 1-34, described growth hormone are pig growth hormone, bovine growth hormone or human growth hormone.

7. Emulsion according to claim 1, its satisfy in following (1)-(3) each or multinomial:

(1) described water is to be selected from water, normal saline and the water buffer one or more;

(2) described oil phase is to be selected from medium chain length fatty acid triglyceride, soybean oil, Semen Maydis oil, Oleum Arachidis hypogaeae semen, oleic acid, PEG400, glycerol, ethanol and the propylene glycol one or more;

(3) described emulsifying agent is for being selected from soybean lecithin, Ovum Gallus domesticus Flavus lecithin, poloxamer 188, poloxamer 407, carbomer, polysorbas20, Tween 80, polyethyleneglycol-12-hydroxy stearin, polyoxyethylene ether (35) Oleum Ricini, polyoxyethylene ether (40) castor oil hydrogenated, polyglycereol-6-dioleate, polyglycereol-3-oleate, Polyethylene Glycol-32-glyceryl laurate ester, Polyethylene Glycol-32-tristerin, and in the Polyethylene Glycol caprylic/capric glyceride one or more.

8. Emulsion according to claim 1, it also comprises the pH regulator agent, and described pH value regulator is selected from one or more in hydrochloric acid, lactic acid, acetic acid, citric acid, phosphoric acid, sodium hydroxide, sodium carbonate, sodium bicarbonate, sodium hydrogen phosphate and the sodium dihydrogen phosphate.

9. according to each described Emulsion in the claim 1 to 8, it is a microemulsion.

10. Emulsion according to claim 1, its component and content are as follows:

Recombinant human insulin 0.03-0.05 weight portion

Sodium hydroxide is an amount of

Deionized water 0.8-1.2 weight portion

Dl-alpha-tocopherol acetate 0.8-1.2 weight portion

Medium chain length fatty acid triglyceride 0.6-0.7 weight portion

Polyethylene Glycol caprylic/capric glyceride 2.5-3.5 weight portion

Poloxamer 188 0.1-0.3 weight portions

Soybean lecithin 0.4-0.6 weight portion;

Particularly, the component of described Emulsion and content are as follows:

Recombinant human insulin's 0.04 weight portion

Sodium hydroxide is an amount of

Deionized water 1 weight portion

Dl-alpha-tocopherol acetate 1 weight portion

Medium chain length fatty acid triglyceride 0.5 weight portion

Polyethylene Glycol caprylic/capric glyceride 3 weight portions

Poloxamer 188 0.2 weight portions

Soybean lecithin 0.5 weight portion.

11. Emulsion according to claim 1, its component and content are as follows:

PTH1-34 0.005-0.015 weight portion

Water 0.4-0.6 weight portion

Acetic acid is an amount of

Dl-alpha-tocopherol 0.6-0.8 weight portion

D-alpha-tocopherol succinic acid macrogol ester 0.4-0.6 weight portion

Soybean oil 1.0-1.4 weight portion

Soybean lecithin 0.6-0.8 weight portion

Polyoxyethylene ether (40) castor oil hydrogenated 2.5-3.5 weight portion

PEG400 1.5-2.5 weight portion;

Particularly, the component of described Emulsion and content are as follows:

PTH1-34 0.01 weight portion

Water 0.5 weight portion

Acetic acid is an amount of

Dl-alpha-tocopherol 0.7 weight portion

D-alpha-tocopherol succinic acid macrogol ester 0.5 weight portion

Soybean oil 1.2 weight portions

Soybean lecithin 0.7 weight portion

Polyoxyethylene ether (40) castor oil hydrogenated 3 weight portions

PEG400 2 weight portions.

12. each described preparation method that contains the Emulsion of hydrophilic biomacromolecule comprises the steps: in the claim 1 to 11

13. the purposes of the described Emulsion of claim 10 in the medicine of preparation treatment or prevent diabetes or blood sugar lowering.

14. the purposes of the described Emulsion of claim 11 in the medicine of preparation treatment or prevention of osteoporosis.

15. the esters derivative of vitamin E and/or vitamin E contains purposes in the Emulsion of hydrophilic biomacromolecule in preparation.