CN110642935A - Tirapapotide analogue - Google Patents

Tirapapotide analogue Download PDF

Info

Publication number
CN110642935A
CN110642935A CN201910908450.3A CN201910908450A CN110642935A CN 110642935 A CN110642935 A CN 110642935A CN 201910908450 A CN201910908450 A CN 201910908450A CN 110642935 A CN110642935 A CN 110642935A
Authority
CN
China
Prior art keywords
ser
gly
ala
pro
analog
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Withdrawn
Application number
CN201910908450.3A
Other languages
Chinese (zh)
Inventor
周述靓
王鹏
邓岚
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Chengdu Aoda Biotechnology Co Ltd
Original Assignee
Chengdu Aoda Biotechnology Co Ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Chengdu Aoda Biotechnology Co Ltd filed Critical Chengdu Aoda Biotechnology Co Ltd
Priority to CN201910908450.3A priority Critical patent/CN110642935A/en
Publication of CN110642935A publication Critical patent/CN110642935A/en
Priority to CN202010247376.8A priority patent/CN111320683B/en
Withdrawn legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/575—Hormones
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/14—Prodigestives, e.g. acids, enzymes, appetite stimulants, antidyspeptics, tonics, antiflatulents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/04—Anorexiants; Antiobesity agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
    • A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A61P9/12—Antihypertensives
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/575—Hormones
    • C07K14/605—Glucagons
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides

Abstract

The invention relates to the field of medicine synthesis, and discloses a tirapapeptide analogue. The tirapapeptide analogues of the present invention are useful for the preparation of a pharmaceutical composition for the treatment of a disease comprising type II diabetes, impaired glucose tolerance, type I diabetes, obesity, hypertension, metabolic syndrome, dyslipidemia, cognitive disorders, atherosclerosis, myocardial infarction, coronary heart disease, cardiovascular disease, stroke, inflammatory bowel syndrome and/or dyspepsia or gastric ulcer, liver fibrosis disease and pulmonary fibrosis.

Description

Tirapapotide analogue
Technical Field
The invention relates to a tirapapeptide analogue and application thereof, wherein the analogue is a dual incretin peptide analogue compound for exciting human glucose-dependent insulinotropic polypeptide (GIP) and glucagon-like peptide-1 (GLP-1) receptors.
Background
GIP is a 42 amino acid gastrointestinal regulatory peptide that plays a physiological role in glucose homeostasis by stimulating insulin secretion from and protecting pancreatic beta cells in the presence of glucose. GLP-1 is a 37 amino acid peptide that stimulates insulin secretion, protects pancreatic beta cells, and inhibits glucagon secretion, gastric emptying, and food intake, resulting in weight loss. GIP and GLP-1 are known as incretins; incretin receptor signaling plays a key physiologically relevant role in glucose homeostasis. In normal physiology, GIP and GLP-1 are secreted from the intestinal tract after a meal, and these incretins enhance physiological responses to food, including satiety, insulin secretion, and nutrient handling.
The most common side effect of GLP-1 analogs is that administration does not achieve full-effect glycemic control and weight loss, whereas GIP alone has a very modest glucose lowering capacity in type 2 diabetics. Both native GIP and GLP-1 can be rapidly inactivated by the ubiquitous protease DPP IV and therefore can only be used for short-term metabolic control.
It is reported in WO 2013/164483 and WO 2014/192284 that certain GIP/GLP-1 analogs exhibit both GIP and GLP-1 activity and that such analogs can be found to achieve better glycemic control and weight loss.
Disclosure of Invention
The invention provides a tirapapeptide analogue and application thereof, wherein the analogue is a dual incretin peptide analogue compound for exciting human glucose-dependent insulinotropic polypeptide (GIP) and glucagon-like peptide-1 (GLP-1) receptors.
To achieve the above object, the present invention provides, in a first aspect, a compound of structure I, a pharmaceutically acceptable salt, solvate, chelate or non-covalent complex thereof, a prodrug based on the compound, or any mixture thereof.
Tyr-AA1-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Tyr-Ser-Ile-AA1-Leu-
Asp-Lys-Ile-Ala-Gln-Lys(R)-Ala-AA2-Val-Gln-Trp-Leu-Ile-Ala-
Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser-AA3
Structure I
AA in Structure I1Comprises the following steps:
Figure BSA0000190780960000021
wherein X1And X2Is a compound of formula (I) in the formula (H),
or is CH3、CH(CH3)2、C(CH3)3、CH(CH2CH3)2、C(CH2CH3)3、CH(CH2CH2CH3)2、C(CH2CH2CH3)3、CH(CH(CH3))2、C(CH(CH3))3、
Or is CH2CH3、CH2CH(CH3)2、CH2C(CH3)3、CH2CH(CH2CH3)2、CH2C(CH2CH3)3、CH2CH(CH2CH2CH3)2、CH2C(CH2CH2CH3)3、CH2CH(CH(CH3))2、CH2C(CH(CH3))3、
AA in Structure I2Is Phe, or is 1-Nal, or is 2-Nal
AA in Structure I3Is NH2Or is OH
R in structure I is HO2C(CH2)n1CO-(γGlu)n2-(PEGn3(CH2)n4CO)n5-
Wherein: n1 is an integer from 10 to 20;
n2 is an integer from 1 to 5;
n3 is an integer from 1 to 30;
n4 is an integer from 1 to 5;
n5 is an integer from 1 to 5;
the invention also provides pharmaceutical compositions comprising a compound according to the invention and the use of a pharmaceutical composition comprising a compound of the invention for the preparation of a medicament for the treatment of a disease.
Preferably, the use of the pharmaceutical composition in the manufacture of a medicament for the treatment of at least one of type II diabetes, impaired glucose tolerance, type I diabetes, obesity, hypertension, metabolic syndrome, dyslipidemia, cognitive disorders, atherosclerosis, myocardial infarction, coronary heart disease, cardiovascular disease, stroke, inflammatory bowel syndrome and/or dyspepsia or gastric ulcer, liver fibrosis diseases and pulmonary fibrosis diseases.
Preferably, the pharmaceutical composition is applied to the preparation of medicines for treating delayed drug effect and/or preventing worsening of type II diabetes.
Preferably, the use of the pharmaceutical composition for the manufacture of a medicament for reducing food intake, reducing beta cell apoptosis, increasing islet beta cell function, increasing beta cell mass, and/or restoring glucose sensitivity to beta cells.
The invention still further provides methods of administering the compounds to a subject for the modulation of blood glucose in vivo.
Further details of the invention are set forth below, or some may be appreciated in embodiments of the invention.
Unless otherwise indicated, the amounts of the various ingredients, reaction conditions, and the like used herein are to be construed in any case to mean "about". Accordingly, unless expressly stated otherwise, all numerical parameters set forth in the following specification and attached claims are approximations that may vary depending upon the standard deviation found in the respective experimental conditions.
Herein, when a chemical structural formula and a chemical name of a compound are ambiguous or ambiguous, the compound is exactly defined by the chemical structural formula. The compounds described herein may contain one or more chiral centers, and/or double bonds and the like, and stereoisomers, including isomers of double bonds (e.g., geometric isomers), optical enantiomers, or diastereomers, may also be present. Accordingly, any chemical structure within the scope of the description, whether partial or complete, including similar structures as described above, includes all possible enantiomers and diastereomers of the compound, including any stereoisomer alone (e.g., pure geometric isomers, pure enantiomers, or pure diastereomers), as well as any mixture of such stereoisomers. Mixtures of these racemates and stereoisomers may also be further resolved into the enantiomers or stereoisomers of their constituent members by those skilled in the art using non-stop separation techniques or methods of chiral molecular synthesis.
The compounds of formula I include, but are not limited to, optical isomers, racemates and/or other mixtures of these compounds. In the above case, a single enantiomer or diastereomer, such as an optical isomer, can be obtained by asymmetric synthesis or racemate resolution. Resolution of the racemates can be accomplished by various methods, such as conventional recrystallization from resolution-assisting reagents, or by chromatographic methods. In addition, the compounds of formula I also include cis and/or trans isomers with double bonds.
The compounds of the present invention include, but are not limited to, the compounds of formula I and all of their pharmaceutically acceptable different forms. The pharmaceutically acceptable different forms of these compounds include various pharmaceutically acceptable salts, solvates, complexes, chelates, non-covalent complexes, prodrugs based on the above and any mixtures of these forms.
The compound shown in the structure I provided by the invention has stable property, is not easily degraded by dipeptidyl peptidase IV (DPP-IV) in vivo, is a GIP/GLP-I dual agonist analogue, and has obvious effects of reducing blood sugar and weight.
Detailed Description
The invention discloses a GIP/GLP-1 analogue and application thereof, and a person skilled in the art can appropriately improve related parameters by referring to the content. It is expressly intended that all such similar substitutes and modifications which would be obvious to one skilled in the art are deemed to be included in the invention. While the process of the present invention has been described in terms of preferred embodiments, it will be apparent to those of ordinary skill in the art that variations and modifications of the compounds and processes described herein, as well as other changes and combinations of the foregoing, may be made to implement and use the techniques of the present invention without departing from the spirit and scope of the invention.
The Chinese names corresponding to the English abbreviations related in the invention are shown in the following table:
english abbreviation Name of Chinese English abbreviation Name of Chinese
Fmoc 9-fluorenylmethoxycarbonyl group OtBu Tert-butoxy radical
tBu Tert-butyl radical Boc Boc-acyl
Trt Trityl radical Pbf (2, 3-dihydro-2, 2, 4, 6, 7-pentamethylbenzofuran-5-yl) sulfonyl group
Ala Alanine Leu Leucine
Arg Arginine Lys Lysine
Asn Asparagine Met Methionine
Asp Aspartic acid Phe Phenylalanine
Cys Cysteine Pro Proline
Gln Glutamine Ser Serine
Glu Glutamic acid Thr Threonine
Gly Glycine Trp Tryptophan
His Histidine Tyr Tyrosine
Ile Isoleucine Val Valine
Dap 2, 3-diaminopropionic acid Dab 2, 4-diaminobutyric acid
Orn Ornithine Dah 2, 7-Diaminoheptanoic acid
Dhthr Dehydroxythreonine Dhval 2, 3-didehydro valine
EXAMPLE 1 preparation of Compound 1
Tyr-Dhthr-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Tyr-Ser-Ile-Dhthr-
Leu-Asp-Lys-Ile-Ala-Gln-Lys (AEEA-AEEA-gamma Glu-18 alkanedioic acid) -
Ala-Phe-Val-Gln-Trp-Leu-Ile-Ala-Gly-Gly-Pro-Ser-Ser-Gly-
Ala-Pro-Pro-Pro-Ser-NH2
The preparation method comprises the following steps: preparing peptide resin by adopting a solid-phase polypeptide synthesis method, carrying out acidolysis on the peptide resin to obtain a crude product, and finally purifying the crude product to obtain a pure product; the step of preparing the peptide resin by the solid-phase polypeptide synthesis method is to sequentially insert corresponding protective amino acids or fragments in the following sequences on a carrier resin by the solid-phase coupling synthesis method to prepare the peptide resin:
in the preparation method, the dosage of the Fmoc-protected amino acid or the protected amino acid fragment is 1.2-6 times of the total mole number of the charged resin; preferably 2.5 to 3.5 times.
In the preparation method, the substitution value of the carrier resin is 0.2-1.0 mmol/g resin, and the preferable substitution value is 0.3-0.5 mmol/g resin.
In a preferred embodiment of the present invention, the solid-phase coupling synthesis method comprises: and (3) after the Fmoc protecting group of the protected amino acid-resin obtained in the previous step is removed, carrying out coupling reaction with the next protected amino acid. The deprotection time for removing Fmoc protection is 10-60 minutes, and preferably 15-25 minutes. The coupling reaction time is 60-300 minutes, and preferably 100-140 minutes.
The coupling reaction needs to add a condensation reagent, and the condensation reagent is selected from one of DIC (N, N-diisopropyl carbodiimide), N, N-dicyclohexylcarbodiimide, benzotriazole-1-yl-oxy tripyrrolidinophosphonium hexafluorophosphate, 2- (7-aza-1H-benzotriazole-1-yl) -1, 1, 3, 3-tetramethylurea hexafluorophosphate, benzotriazole-N, N, N ', N' -tetramethylurea hexafluorophosphate or O-benzotriazole-N, N, N ', N' -tetramethylurea tetrafluoroborate; n, N-diisopropylcarbodiimide is preferred. The molar consumption of the condensation reagent is 1.2-6 times of the total molar number of amino groups in the amino resin, and preferably 2.5-3.5 times.
The coupling reaction needs to add an activating reagent, wherein the activating reagent is selected from 1-hydroxybenzotriazole or N-hydroxy-7-azabenzotriazole, and 1-hydroxybenzotriazole is preferred. The amount of the activating agent is 1.2 to 6 times, preferably 2.5 to 3.5 times of the total mole number of the amino groups in the amino resin.
As a preferable scheme of the invention, the reagent for removing Fmoc protection is PIP/DMF (piperidine/N, N-dimethylformamide) mixed solution, and the piperidine content in the mixed solution is 10-30% (V). The dosage of the Fmoc protection removing reagent is 5-15 mL per gram of amino resin, and preferably 8-12 mL per gram of amino resin.
Preferably, the peptide resin is subjected to acidolysis while removing the resin and side chain protecting groups to obtain a crude product:
more preferably, the acidolysis agent used in the acidolysis of the peptide resin is a mixed solvent of trifluoroacetic acid (TFA), 1, 2-Ethanedithiol (EDT) and water, and the volume ratio of the mixed solvent is as follows: 80-95% of TFA, 1-10% of EDT and the balance of water.
More preferably, the volume ratio of the mixed solvent is: 89-91% of TFA, 4-6% of EDT and the balance of water. Optimally, the volume ratio of the mixed solvent is as follows: TFA 90%, EDT 5%, balance water.
The dosage of the acidolysis agent is 4-15 mL per gram of the peptide resin; preferably, 7-10 mL of acidolysis agent is required per gram of peptide resin.
The time for cracking by using the acidolysis agent is 1-6 hours, preferably 3-4 hours at room temperature.
Further, the crude product is purified by high performance liquid chromatography and freeze-dried to obtain a pure product.
1. Synthesis of peptide resins
Rink Amide BHHA resin is used as carrier resin, and is coupled with protected amino acid shown in the following table in sequence through Fmoc protection removal and coupling reaction to prepare peptide resin. The protected amino acids used in this example correspond to the protected amino acids shown below:
Figure BSA0000190780960000061
(1) 1 st protected amino acid inserted into main chain
Dissolving 0.03mol of the 1 st protected amino acid and 0.03mol of HOBt in a proper amount of DMF; and adding 0.03mol DIC slowly into the protected amino acid DMF solution under stirring, and reacting for 30 minutes under stirring at room temperature to obtain an activated protected amino acid solution for later use.
0.01mol of Rink amide MBHA resin (substitution value about 0.4mmol/g) is taken, deprotected by 20% PIP/DMF solution for 25 minutes, washed and filtered to obtain Fmoc-removed resin.
And adding the activated 1 st protected amino acid solution into the Fmoc-removed resin, performing coupling reaction for 60-300 minutes, and filtering and washing to obtain the resin containing 1 protected amino acid.
(2) 2-40 protected amino acids connected to main chain
And sequentially inoculating the corresponding 2 nd to 40 th protected amino acids by the same method for inoculating the 1 st protected amino acid of the main chain to obtain the resin containing 40 amino acids of the main chain.
(3) Side chain insertion of the 1 st protected amino acid
Dissolving 0.03mol of the 1 st protected amino acid of the side chain and 0.03mol of HOBt in a proper amount of DMF; and adding 0.03mol DIC slowly into the protected amino acid DMF solution under stirring, and reacting for 30 minutes under stirring at room temperature to obtain the activated protected amino acid solution.
Taking 2.5mmol of tetratriphenylphosphine palladium and 25mmol of phenylsilane, dissolving with a proper amount of dichloromethane, deprotecting for 4 hours, filtering and washing to obtain a resin without Alloc for later use.
Adding the activated side chain 1 st protected amino acid solution into the Alloc-removed resin, performing coupling reaction for 60-300 minutes, filtering and washing to obtain the side chain 1 st protected amino acid-containing resin.
(4) 2-4 protective amino acids of grafted side chain
And sequentially inoculating 2 nd to 4 th protected amino acids and single protected fatty acids corresponding to side chains by adopting the same method for inoculating the 1 st protected amino acid to the main chain to obtain the peptide resin.
2. Preparation of crude product
Taking the peptide resin, adding a cracking reagent (10 mL/g resin of the cracking reagent) with the volume ratio of TFA, water and EDT being 95: 5, uniformly stirring, stirring at room temperature for reaction for 3 hours, filtering a reaction mixture by using a sand core funnel, collecting filtrate, washing the resin with a small amount of TFA for 3 times, combining the filtrates, concentrating under reduced pressure, adding anhydrous ether for precipitation, washing the precipitate with anhydrous ether for 3 times, and drying to obtain white-like powder, namely a crude product.
3. Preparation of the pure product
Mixing the crude product with water, stirring, adjusting pH to 8.0 with ammonia water to dissolve completely, filtering the solution with 0.45 μm mixed microporous membrane, and purifying;
purifying by high performance liquid chromatography, wherein the chromatographic packing for purification is 10 μm reversed phase C18, the mobile phase system is 0.1% TFA/water solution-0.1% TFA/acetonitrile solution, the flow rate of a 30mm by 250mm chromatographic column is 20mL/min, eluting by a gradient system, circularly sampling for purification, sampling the crude product solution in the chromatographic column, starting the mobile phase for elution, collecting the main peak, and evaporating acetonitrile to obtain a purified intermediate concentrated solution;
filtering the purified intermediate concentrated solution with 0.45 μm filter membrane for use, and performing salt exchange by high performance liquid chromatography with 1% acetic acid/water solution-acetonitrile as mobile phase system, 10 μm reversed phase C18 as purification chromatographic filler, and 20mL/min of 30 mm/250 mm chromatographic column flow rate (corresponding flow rate can be adjusted according to chromatographic columns of different specifications); adopting gradient elution and circulation sample loading method, loading sample in chromatographic column, starting mobile phase elution, collecting atlas, observing change of absorbance, collecting main peak of salt exchange and analyzing liquid phase to detect purity, combining main peak solutions of salt exchange, concentrating under reduced pressure to obtain pure acetic acid water solution, and freeze drying to obtain pure product 5.5g, purity of 97.5% and total yield of 11.4%. The molecular weight was 4837.6 (100% M + H).
EXAMPLE 2 preparation of Compound 2
Tyr-Dhval-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Tyr-Ser-Ile-Dhval-
Leu-Asp-Lys-Ile-Ala-Gln-Lys (AEEA-AEEA-gamma-Glu-18 alkanedioic acid) -
Ala-Phe-Val-Gln-Trp-Leu-Ile-Ala-Gly-Gly-Pro-Ser-Ser-Gly-
Ala-Pro-Pro-Pro-Ser-NH2
The procedure is as in example 1, using the protected amino acids as in the following table:
Figure BSA0000190780960000091
Figure BSA0000190780960000101
6.1g of pure product is obtained, the purity is 96.9 percent, and the total yield is 12.5 percent. The molecular weight was 4865.2 (100% M + H).
EXAMPLE 3 preparation of Compound 3
Tyr-Dhthr-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Tyr-Ser-Ile-Dhthr-
Leu-Asp-Lys-Ile-Ala-Gln-Lys(PEG5CH2CO-gamma Glu-18 alkanedioic acid)
Ala-Phe-Val-Gln-Trp-Leu-Ile-Ala-Gly-Gly-Pro-Ser-Ser-Gly-
Ala-Pro-Pro-Pro-Ser-NH2
The procedure is as in example 1, using the protected amino acids as in the following table:
Figure BSA0000190780960000121
7.4g of pure product is obtained, the purity is 96.3 percent, and the total yield is 15.3 percent. The molecular weight was 4824.8 (100% M + H).
EXAMPLE 4 preparation of Compound 4
Tyr-Dhval-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Tyr-Ser-Ile-Dhval-
Leu-Asp-Lys-Ile-Ala-Gln-Lys(PEG5CH2CO-gamma Glu-18 alkanedioic acid)
Ala-Phe-Val-Gln-Trp-Leu-Ile-Ala-Gly-Gly-Pro-Ser-Ser-Gly-
Ala-Pro-Pro-Pro-Ser-NH2
The procedure is as in example 1, using the protected amino acids as in the following table:
Figure BSA0000190780960000131
Figure BSA0000190780960000141
7.1g of pure product is obtained, the purity is 96.8 percent, and the total yield is 14.6 percent. The molecular weight was 4852.4 (100% M + H).
Example 7 Activity assay
1. Measurement method
GLP-1R is mainly present on the surface of islet beta cells and is a G protein-coupled receptor (GPCRs). GLP-1R, stimulated by its specific agonist, activates the intracellular adenylate cyclase pathway, raises cAMP levels, and ultimately leads to the production and release of insulin. A cell strain which is stably transfected with GLP-1R is stimulated by an object to be tested to ensure that the intracellular cAMP level of the cell is rapidly increased, and the Relative Light Unit (RLU) after each dose of stimulated cells is measured by a chemiluminescence method to further calculate the EC50 of the agonist, wherein the activity measuring method is a GLP-1 receptor agonist activity measuring method which is commonly used at home and abroad at present.
The CHO-K1 cell strain capable of stably expressing GLP-1R is adopted, agonist with different concentrations is used for stimulating stable cells, and the relative light unit of the stimulated cells is measured to obtain the bioactivity of the agonist.
2. Measurement results
The results of the assay are shown in the following table:
compound numbering Biological Activity (%)
Compound 1 121.72
Compound 2 66.57
Compound 3 70.25
Compound 4 55.81
EXAMPLE 8 determination of Primary pharmacokinetic Properties
Each compound was divided into two dosing groups: SD rats, 4 males per group, 8 in total.
Tail vein intravenous injection group: the dose is 1mg/kg, rat orbital veins are respectively bled before (0h) and 30min, 1h, 2h, 4h, 8h, 24h, 48h, 96h and 144h after administration, and plasma samples are centrifugally separated.
Subcutaneous administration group: the dose is 1mg/kg, rat orbital veins are respectively bled before (0h) and 1h, 2h, 3h, 4h, 8h, 24h, 48h, 96h and 144h after administration, and plasma samples are separated by centrifugation.
Plasma concentrations of the corresponding compounds in plasma samples of SD rats were measured by the liquid chromatography-mass spectrometry method, and the half-lives of the compounds after intravenous and subcutaneous administration in SD rats under Subcutaneous (SC) administration are shown in the following table:
compound (I) t1/2(h)
Compound 1 9.1
Compound 2 8.7
Compound 3 9.5
Compound 4 9.3

Claims (7)

1. A tirapatide analog having structural formula I:
Tyr-AA1-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Tyr-Ser-Ile-AA1-Leu-
Asp-Lys-Ile-Ala-Gln-Lys(R)-Ala-AA2-Val-Gln-Trp-Leu-Ile-Ala-
Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser-AA3
structure I
AA in Structure I1Comprises the following steps:
Figure FSA0000190780950000011
wherein X1And X2Is a compound of formula (I) in the formula (H),
or is CH3、CH(CH3)2、C(CH3)3、CH(CH2CH3)2、C(CH2CH3)3、CH(CH2CH2CH3)2、C(CH2CH2CH3)3、CH(CH(CH3))2、C(CH(CH3))3、
Or is CH2CH3、CH2CH(CH3)2、CH2C(CH3)3、CH2CH(CH2CH3)2、CH2C(CH2CH3)3、CH2CH(CH2CH2CH3)2、CH2C(CH2CH2CH3)3、CH2CH(CH(CH3))2、CH2C(CH(CH3))3、
AA in Structure I2Is Phe, or is 1-Nal, or is 2-Nal
AA in Structure I3Is NH2Or is OH
R in structure I is HO2C(CH2)n1CO-(γGlu)n2-(PEGn3(CH2)n4CO)n5-
Wherein: n1 is an integer from 10 to 20;
n2 is an integer from 1 to 5;
n3 is an integer from 1 to 30;
n4 is an integer from 1 to 5;
n5 is an integer from 1 to 5.
2. A tirapatide analog according to claim 1, comprising a pharmaceutically acceptable salt, solvate, chelate or non-covalent complex of the analog, a prodrug based on the compound, or a mixture of any of the foregoing.
3. A tirapapeptide analog according to claim 1 and claim 2 for use in the preparation of a pharmaceutical composition for the treatment of a disease.
4. The pharmaceutical composition according to claim 3, for use in the manufacture of a medicament for the treatment of at least one of type II diabetes, impaired glucose tolerance, type I diabetes, obesity, hypertension, metabolic syndrome, dyslipidemia, cognitive disorders, atherosclerosis, myocardial infarction, coronary heart disease, cardiovascular disease, stroke, inflammatory bowel syndrome and/or dyspepsia or gastric ulcer, liver fibrosis diseases and pulmonary fibrosis diseases.
5. The pharmaceutical composition according to claim 4, for use in the preparation of a medicament for the treatment of delayed efficacy and/or prevention of exacerbation of type II diabetes.
6. The pharmaceutical composition of claim 6, for use in the preparation of a medicament for reducing food intake, reducing beta cell apoptosis, increasing islet beta cell function, increasing beta-cell mass, and/or restoring glucose sensitivity to beta cells.
7. The tirapapeptide analog of claim 1, comprising the analog for use in a method of modulating blood glucose in vivo.
CN201910908450.3A 2019-09-25 2019-09-25 Tirapapotide analogue Withdrawn CN110642935A (en)

Priority Applications (2)

Application Number Priority Date Filing Date Title
CN201910908450.3A CN110642935A (en) 2019-09-25 2019-09-25 Tirapapotide analogue
CN202010247376.8A CN111320683B (en) 2019-09-25 2020-03-31 Tilappa peptide analogue

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201910908450.3A CN110642935A (en) 2019-09-25 2019-09-25 Tirapapotide analogue

Publications (1)

Publication Number Publication Date
CN110642935A true CN110642935A (en) 2020-01-03

Family

ID=69011191

Family Applications (2)

Application Number Title Priority Date Filing Date
CN201910908450.3A Withdrawn CN110642935A (en) 2019-09-25 2019-09-25 Tirapapotide analogue
CN202010247376.8A Active CN111320683B (en) 2019-09-25 2020-03-31 Tilappa peptide analogue

Family Applications After (1)

Application Number Title Priority Date Filing Date
CN202010247376.8A Active CN111320683B (en) 2019-09-25 2020-03-31 Tilappa peptide analogue

Country Status (1)

Country Link
CN (2) CN110642935A (en)

Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111320683A (en) * 2019-09-25 2020-06-23 成都奥达生物科技有限公司 Tirapapotide analogue
CN111333714A (en) * 2020-03-05 2020-06-26 成都奥达生物科技有限公司 Long-acting GLP-1 compound
WO2022117056A1 (en) * 2020-12-02 2022-06-09 南京明德新药研发有限公司 Lactam-modified polypeptide compounds
CN116162146A (en) * 2021-11-24 2023-05-26 成都奥达生物科技有限公司 GIP-GLP-1 double-agonist compound
CN116854805A (en) * 2023-09-05 2023-10-10 杭州湃肽生化科技有限公司 Preparation method of telipopeptide
US11779648B2 (en) 2020-07-22 2023-10-10 Novo Nordisk A/S Co-agonists at GLP-1 and GIP receptors suitable for oral delivery
US11840560B2 (en) 2022-01-20 2023-12-12 Novo Nordisk A/S Prodrugs and uses thereof
CN115124602B (en) * 2021-03-25 2024-01-26 博瑞生物医药（苏州）股份有限公司 Dual receptor agonists of GIP and GLP-1, pharmaceutical compositions and uses

Families Citing this family (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN116710462A (en) 2021-01-20 2023-09-05 维京治疗公司 Compositions and methods for treating metabolic disorders and liver diseases
CN116847868A (en) * 2021-01-29 2023-10-03 韩美药品株式会社 Pharmaceutical composition comprising GIP derivative or long-acting conjugate thereof for preventing or treating pulmonary diseases
CN116023444B (en) * 2022-08-12 2023-08-01 重庆宸安生物制药有限公司 GIP and GLP-1 dual-receptor agonist polypeptide compound and application thereof

Family Cites Families (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
TWI829687B (en) * 2018-05-07 2024-01-21 丹麥商諾佛 儂迪克股份有限公司 Solid compositions comprising a glp-1 agonist and a salt of n-(8-(2-hydroxybenzoyl)amino)caprylic acid
TWI705820B (en) * 2018-06-22 2020-10-01 美商美國禮來大藥廠 Gip/glp1 agonist compositions
EP3826716A1 (en) * 2018-07-23 2021-06-02 Eli Lilly and Company Methods of using a gip/glp1 co-agonist for therapy
TWI799680B (en) * 2019-01-29 2023-04-21 美商美國禮來大藥廠 Process for preparing a gip/glp1 dual agonist
CN110642935A (en) * 2019-09-25 2020-01-03 成都奥达生物科技有限公司 Tirapapotide analogue

Cited By (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111320683A (en) * 2019-09-25 2020-06-23 成都奥达生物科技有限公司 Tirapapotide analogue
CN111320683B (en) * 2019-09-25 2023-06-27 成都奥达生物科技有限公司 Tilappa peptide analogue
CN111333714A (en) * 2020-03-05 2020-06-26 成都奥达生物科技有限公司 Long-acting GLP-1 compound
US11779648B2 (en) 2020-07-22 2023-10-10 Novo Nordisk A/S Co-agonists at GLP-1 and GIP receptors suitable for oral delivery
WO2022117056A1 (en) * 2020-12-02 2022-06-09 南京明德新药研发有限公司 Lactam-modified polypeptide compounds
CN115124602B (en) * 2021-03-25 2024-01-26 博瑞生物医药（苏州）股份有限公司 Dual receptor agonists of GIP and GLP-1, pharmaceutical compositions and uses
CN116162146A (en) * 2021-11-24 2023-05-26 成都奥达生物科技有限公司 GIP-GLP-1 double-agonist compound
CN116162146B (en) * 2021-11-24 2023-10-24 成都奥达生物科技有限公司 GIP-GLP-1 double-agonist compound
US11840560B2 (en) 2022-01-20 2023-12-12 Novo Nordisk A/S Prodrugs and uses thereof
CN116854805A (en) * 2023-09-05 2023-10-10 杭州湃肽生化科技有限公司 Preparation method of telipopeptide
CN116854805B (en) * 2023-09-05 2023-12-15 杭州湃肽生化科技有限公司 Preparation method of telipopeptide

Also Published As

Publication number Publication date
CN111320683B (en) 2023-06-27
CN111320683A (en) 2020-06-23

Similar Documents

Publication Publication Date Title
CN111320683B (en) Tilappa peptide analogue
CN110590934B (en) GLP-1 compound
CN110551203B (en) Exenatide analogue
CN111333714A (en) Long-acting GLP-1 compound
CN111423506B (en) GLP-1 compound
WO2012155780A1 (en) Branched-peg modified glp-1 analogue and pharmaceutically acceptable salts thereof
CN107253985B (en) Design and application of long-acting hypoglycemic peptide
CN116162146B (en) GIP-GLP-1 double-agonist compound
CN114075275A (en) Long-acting insulin analogue
CN115594752A (en) Long-acting double-agonist compound
CN111285923A (en) PSD-95 inhibitor
CN107298708B (en) Glucagon-like peptide-1 (GLP-1) analogue with ether bond and application thereof
CN109694404B (en) Hypoglycemic peptide and application thereof
CN115594753A (en) Long-acting double-agonist compound
CN117586373A (en) Long-acting dual agonist compound
CN117229364A (en) GIP-GLP-1 double-agonist compound
CN117229365A (en) GIP-GLP-1 double-agonist compound
WO2023155545A1 (en) Long-acting glp-1/glp-2 dual agonist compound
CN114478694A (en) Long-acting MC4R agonist
CN116284329B (en) Long-acting natriuretic peptide compound
CN116162147B (en) Long-acting insulin analogue
CN109824771B (en) Hypoglycemic peptide and application thereof
WO2024032454A1 (en) Long-acting abaloparatide compound
WO2024032457A1 (en) Long-acting teriparatide compound
CN115043904A (en) Long-acting K opioid receptor agonist

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
WW01 Invention patent application withdrawn after publication
WW01 Invention patent application withdrawn after publication

Application publication date: 20200103