WO2022117056A1 - 含内酰胺修饰的多肽类化合物 - Google Patents
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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
- C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
- C07K14/72—Receptors; Cell surface antigens; Cell surface determinants for hormones
-
- 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
-
- 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
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/22—Hormones
- A61K38/26—Glucagons
-
- 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
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
Definitions
- the present invention relates to a class of lactam-modified polypeptide compounds and their application in preparing medicines for treating related diseases.
- GIP glucose-dependent insulinotropic peptide
- GLP-1 glucagon-like peptide-1
- GLP-1 plays a role in protecting islet beta cells in islets, stimulates islet beta cells to release insulin in a glucose-dependent manner, and effectively controls postprandial blood sugar. Due to its unique mechanism of action, the risk of hypoglycemia is greatly reduced.
- GLP-1R agonists have shown excellent hypoglycemic effects in clinical practice, there are still many patients with type 2 diabetes who fail to achieve their hypoglycemic and weight loss goals. Therefore, there is an urgent and promising need to combine GLP-1R agonists with other hypoglycemic targets such as GIP for the treatment of type 2 diabetes.
- GIP is a polypeptide secreted by neuroendocrine K cells in the small intestine, and its physiological effects are mediated by GIPR, mainly including glucose-independent insulin secretion, glucagon secretion, and lipid metabolism. While the beneficial effects of GIPR agonists appear to be attenuated in hyperglycemic symptoms in patients with type 2 diabetes, studies have shown that the attenuated insulinotropic effect of GIP can be fully restored after a period of normalization of plasma glucose levels. This indicated that co-agonism of GLP-1R/GIPR could exert a synergistic hypoglycemic effect.
- the dual agonism of GIP and GLP-1 has a synergistic effect on the regulation of glucose/lipid metabolism, and has better therapeutic effects on lowering blood sugar, reducing body weight and relieving liver fat.
- the present invention provides a compound of the formula or a pharmaceutically acceptable salt thereof,
- Z 0 is selected from glutamine (Q) and asparagine (N);
- Z 1 is selected from alanine (A) and glutamic acid (E);
- Z 2 is selected from valine (V) and isoleucine (I);
- Z 3 is selected from isoleucine (I) and leucine (L);
- E 0 and K 0 indicate that the carboxyl group on the glutamic acid side chain and the amino group on the lysine side chain together form a lactam, and the structure of K 0 Z 1 FZ 2 E 0 is:
- the structure of E 0 AQK 0 is The structure of E 0 LDKK 0 is The structure of E 0 LDK 0 is
- K 1 indicates that the amino group on the side chain of lysine is connected with -XX 1 -X 2 , and its structure is
- X is selected from
- R 1 and R 2 are each independently selected from H and CH 3 ;
- X 1 is selected from
- X 2 is selected from
- n and p are independently selected from 2 and 3, respectively;
- s is selected from 2 and 3;
- q is selected from 15, 16, 17, 18 and 19.
- the above-mentioned compound, or a pharmaceutically acceptable salt thereof is selected from:
- Z 0 , Z 1 , Z 2 , Z 3 and K 1 are as defined in the present invention.
- m, n and p are independently selected from 2, and other variables are as defined in the present invention.
- the above s is selected from 2, and other variables are as defined in the present invention.
- the above q is selected from 15 and 17, and other variables are as defined in the present invention.
- X 1 is selected from Other variables are as defined in the present invention.
- the above X 2 is selected from Other variables are as defined in the present invention.
- the above -XX 1 -X 2 are selected from Other variables are as defined in the present invention.
- the present invention provides a compound of the formula or a pharmaceutically acceptable salt thereof,
- K 0 and E 0 indicate that the amino group on the side chain of lysine and the carboxyl group on the side chain of glutamic acid together form a lactam, and its structure is
- K 1 indicates that the amino group on the side chain of lysine is connected with -XX 1 -X 2 , and its structure is
- X is selected from
- R 1 and R 2 are each independently selected from H;
- X 1 is selected from,
- X 2 is selected from
- n and n are independently selected from 2 and 3, respectively;
- s is selected from 2 and 3;
- q is selected from 15, 16, 17, 18 and 19.
- m and n are independently selected from 2, and other variables are as defined in the present invention.
- the above s is selected from 2, and other variables are as defined in the present invention.
- the above q is selected from 15 and 17, and other variables are as defined in the present invention.
- X 1 is selected from Other variables are as defined in the present invention.
- the above X 2 is selected from Other variables are as defined in the present invention.
- the above -XX 1 -X 2 are selected from Other variables are as defined in the present invention.
- the present invention provides a compound of the formula or a pharmaceutically acceptable salt thereof,
- Z 0 is selected from glutamine (Q) and asparagine (N);
- Z 1 is selected from alanine (A) and glutamic acid (E);
- Z 2 is selected from valine (V) and isoleucine (I);
- Z 3 is selected from isoleucine (I) and leucine (L);
- E 0 and K 0 indicate that the carboxyl group on the glutamic acid side chain and the amino group on the lysine side chain together form a lactam, and the structure of K 0 Z 1 FZ 2 E 0 is:
- the structure of E 0 AQK 0 is The structure of E 0 LDKK 0 is
- K 1 indicates that the amino group on the side chain of lysine is connected with -XX 1 -X 2 , and its structure is
- X is selected from
- R 1 and R 2 are each independently selected from H;
- X 1 is selected from
- X 2 is selected from
- n and p are independently selected from 2 and 3, respectively;
- s is independently selected from 2 and 3;
- q is independently selected from 15, 16, 17, 18 and 19.
- m, n and p are independently selected from 2, and other variables are as defined in the present invention.
- the above s are independently selected from 2, and other variables are as defined in the present invention.
- the above q is independently selected from 15 and 17, and other variables are as defined herein.
- the above q is independently selected from 17, and other variables are as defined herein.
- X 1 is selected from Other variables are as defined in the present invention.
- the above X 2 is selected from Other variables are as defined in the present invention.
- the above -XX 1 -X 2 are selected from Other variables are as defined in the present invention.
- the present invention provides a compound of the formula or a pharmaceutically acceptable salt thereof,
- K 0 and E 0 indicate that the amino group on the side chain of lysine and the carboxyl group on the side chain of glutamic acid together form a lactam, and its structure is
- K 1 indicates that the amino group on the side chain of lysine is connected with -XX 1 -X 2 , and its structure is
- X is selected from
- R 1 and R 2 are each independently selected from H;
- X 1 is selected from
- X 2 is selected from
- n and p are independently selected from 2 and 3, respectively;
- q is independently selected from 15, 16, 17, 18 and 19.
- m, n and p are independently selected from 2, and other variables are as defined in the present invention.
- the above q is independently selected from 17, and other variables are as defined herein.
- X 1 is selected from Other variables are as defined in the present invention.
- the above X 2 is selected from Other variables are as defined in the present invention.
- the above -XX 1 -X 2 are selected from
- the present invention provides a compound of the formula or a pharmaceutically acceptable salt thereof,
- E 0 and K 0 indicate that the carboxyl group on the side chain of glutamic acid and the amino group on the side chain of lysine together form a lactam, and its structure is
- K 1 indicates that the amino group on the side chain of lysine is connected with -XX 1 -X 2 , and its structure is
- X is selected from
- R 1 and R 2 are each independently selected from H;
- X 1 is selected from
- X 2 is selected from
- n and p are independently selected from 2 and 3, respectively;
- q is independently selected from 15, 16, 17, 18 and 19.
- m, n and p are independently selected from 2, and other variables are as defined in the present invention.
- the above q is independently selected from 17, and other variables are as defined herein.
- X 1 is selected from Other variables are as defined in the present invention.
- the above X 2 is selected from Other variables are as defined in the present invention.
- the above -XX 1 -X 2 are selected from Other variables are as defined in the present invention.
- the present invention also provides a compound represented by the following formula or a pharmaceutically acceptable salt thereof,
- the present invention also provides a compound represented by the following formula or a pharmaceutically acceptable salt thereof,
- the above compounds or their pharmaceutically acceptable salts are used in the preparation of medicaments for the treatment of obesity and diabetes.
- the present invention also provides following test method:
- CHO-hERG cell line Choinese hamster ovary cells stably expressing hERG channels, constructed in-house by Shanghai Institute of Materia Medica, Chinese Academy of Sciences; Axopatch 200B patch clamp amplifier, Taser International;
- CHO cells stably expressing hERG were cultured in a cell culture dish with a diameter of 35 mm, placed in an incubator at 37°C, 5% CO 2 , and passaged at a ratio of 1:5 every 48 hours.
- the cell culture medium was aspirated, rinsed with extracellular fluid, and then 0.25% Trypsin-EDTA (Invitrogen) solution was added, and the cells were digested at room temperature for 3-5 minutes. Aspirate the digestion solution, resuspend the cells with extracellular fluid, and transfer the cells to the experimental dish for electrophysiological recording.
- Extracellular fluid needs to be prepared once a month.
- the intracellular fluid must be aliquoted and stored at -20°C.
- Extracellular fluid 145NaCl, 4KCl , 2CaCl2 , 1MgCl2, 10Glucose and 10HEPES, pH adjusted to 7.4 with NaOH, osmotic pressure 295mOsm.
- Intracellular solution 120KCl, 31.25KOH, 5.374CaCl2 , 1.75MgCl2 , 4Na2ATP , 10 HEPES and 10EGTA, pH adjusted to 7.2 with KOH, osmotic pressure 285 mOsm.
- the compound was dissolved in DMSO into 20 mM mother solution.
- the compound mother solution was serially diluted 3 times with DMSO, that is, 10 ⁇ L of the compound mother solution was added to 20 ⁇ L of DMSO, and 6 intermediate concentrations of the compounds serially diluted in DMSO were obtained in turn. 20, 6.66, 2.22, 0.74, 0.24 and 0.082 mM.
- 10 ⁇ L of the intermediate concentration of the compound was added to 4990 ⁇ L of extracellular fluid, and the final concentration to be tested was obtained by 500-fold dilution.
- the content of DMSO in the final test concentration did not exceed 0.2%, and this concentration of DMSO had no effect on the hERG potassium channel.
- CHO Choinese Hamster Ovary
- hERG potassium channel currents were recorded by whole-cell patch-clamp technique at room temperature.
- the glass microelectrode is drawn from the glass electrode blank (BF150-86-10, Sutter) by a drawing machine.
- the tip resistance after filling the electrode liquid is about 2-5M ⁇ .
- Clamp voltages and data recording were controlled and recorded by a computer using pClamp 10 software with a sampling frequency of 10 kHz and a filter frequency of 2 kHz.
- the cells were clamped at -80mV and the step voltage evoked hERG potassium current (IhERG) was given a 2s depolarization voltage from -80mV to +20mV, and then repolarized to -50mV for 1s after back to -80mV. This voltage stimulation was given every 10 s, and the dosing process was started after it was determined that the hERG potassium current was stable (1 min).
- Compound concentrations were administered consecutively starting from the low test concentration, and each test concentration was administered for at least 1 minute. Compounds were tested at least 3 cells per concentration (n > 3) and positive compounds were tested at least 2 cells per concentration (n > 2).
- I (C) I b +(I fr -I b )*c n /(IC 50 n +c n )
- c is the compound test concentration
- n is the slope
- Curve fitting and inhibition rate calculation are both completed by Qpatch analysis software. If the inhibition rate at the lowest concentration exceeds half inhibition or the inhibition rate at the highest concentration does not reach half inhibition, the corresponding IC 50 of the compound is lower than the lowest concentration or IC 50 value. greater than the highest concentration.
- the compounds of the present invention are free of risks associated with hERG.
- test compounds The inhibitory effect of test compounds on the activity of human liver microsomal cytochrome P450 isoenzymes (CYP1A2, CYP2C9, CYP2C19, CYP2D6 and CYP3A4) was determined.
- test compound (10mM) was diluted in gradient to prepare working solution (100 ⁇ final concentration), the concentration of working solution was: 5, 1.5, 0.5, 0.15, 0.05, 0.015, 0.005mM, and P450 isoenzyme ( CYP1A2, CYP2C9, CYP2C19, CYP2D6 and CYP3A4) positive inhibitors and their specific substrate mixtures; thawed human liver microsomes frozen at –80°C on ice until all human liver microsomes were dissolved.
- concentration of working solution was: 5, 1.5, 0.5, 0.15, 0.05, 0.015, 0.005mM
- P450 isoenzyme CYP1A2, CYP2C9, CYP2C19, CYP2D6 and CYP3A4
- the compounds of the present invention have no associated risk of CYP inhibition.
- the compound of the present invention has strong agonistic activity on GLP-1R/GIPR; the compound of the present invention has excellent pharmacokinetic properties; the compound of the present invention has excellent plasma stability; the compound of the present invention has a very high degree of plasma protein binding; The compounds of the present invention have excellent in vivo efficacy.
- the term "pharmaceutically acceptable” refers to those compounds, materials, compositions and/or dosage forms which, within the scope of sound medical judgment, are suitable for use in contact with human and animal tissue , without excessive toxicity, irritation, allergic reactions or other problems or complications, commensurate with a reasonable benefit/risk ratio.
- salts refers to salts of the compounds of the present invention, prepared from compounds with specific substituents discovered by the present invention and relatively non-toxic acids or bases.
- base addition salts can be obtained by contacting such compounds with a sufficient amount of base in neat solution or in a suitable inert solvent.
- Pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amine or magnesium salts or similar salts.
- acid addition salts can be obtained by contacting such compounds with a sufficient amount of acid in neat solution or in a suitable inert solvent.
- Examples of pharmaceutically acceptable acid addition salts include inorganic acid salts including, for example, hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, bicarbonate, phosphoric acid, monohydrogen phosphate, dihydrogen phosphate, sulfuric acid, Hydrogen sulfate, hydroiodic acid, phosphorous acid, etc.; and organic acid salts including, for example, acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, Similar acids such as fumaric, lactic, mandelic, phthalic, benzenesulfonic, p-toluenesulfonic, citric, tartaric, and methanesulfonic acids; also include salts of amino acids such as arginine, etc. , and salts of organic acids such as glucuronic acid. Certain specific compounds of the present invention contain both basic and acidic functional groups and thus can be converted into either base
- the pharmaceutically acceptable salts of the present invention can be synthesized from the acid or base containing parent compound by conventional chemical methods. Generally, such salts are prepared by reacting the free acid or base form of these compounds with a stoichiometric amount of the appropriate base or acid in water or an organic solvent or a mixture of the two.
- a or Ala as described herein represents alanine, and the structure is R or Arg means arginine, the structure is N or Asn means asparagine, the structure is D or Asp means aspartic acid, the structure is C or Cys means cysteine, the structure is Q or Gln represents glutamine, the structure is E or Glu means glutamic acid, the structure is G or Gly means glycine, the structure is H or His means histidine, the structure is I or Ile represents isoleucine, the structure is L or Leu means leucine, the structure is K or Lys means lysine, the structure is M or Met stands for methionine, the structure is F or Phe means phenylalanine, the structure is P or Pro means proline, the structure is S or Ser represents serine, the structure is T or Thr means threonine, the structure is W or Trp means tryptophan, the structure is Y or Tyr represents tyrosine, the structure is V or Val means valine, the
- treating includes inhibiting, slowing, stopping or reversing the progression or severity of an existing symptom or condition.
- the term “isomer” is intended to include geometric isomers, cis-trans isomers, stereoisomers, enantiomers, optical isomers, diastereomers and tautomers isomer.
- the compounds of the present invention may exist in specific geometric or stereoisomeric forms.
- the present invention contemplates all such compounds, including cis and trans isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers isomers, (D)-isomers, (L)-isomers, and racemic mixtures thereof and other mixtures, such as enantiomerically or diastereomerically enriched mixtures, all of which belong to this within the scope of the invention.
- Additional asymmetric carbon atoms may be present in substituents such as alkyl groups. All such isomers, as well as mixtures thereof, are included within the scope of the present invention.
- enantiomers or “optical isomers” refer to stereoisomers that are mirror images of each other.
- cis-trans isomer or “geometric isomer” result from the inability to rotate freely due to double bonds or single bonds to ring carbon atoms.
- diastereomer refers to a stereoisomer in which the molecule has two or more chiral centers and the molecules are in a non-mirror-image relationship.
- the terms “enriched in one isomer”, “enriched in isomer”, “enriched in one enantiomer” or “enriched in one enantiomer” refer to one of the isomers or pairs
- the enantiomer content is less than 100%, and the isomer or enantiomer content is greater than or equal to 60%, or greater than or equal to 70%, or greater than or equal to 80%, or greater than or equal to 90%, or greater than or equal to 95%, or Greater than or equal to 96%, or greater than or equal to 97%, or greater than or equal to 98%, or greater than or equal to 99%, or greater than or equal to 99.5%, or greater than or equal to 99.6%, or greater than or equal to 99.7%, or greater than or equal to 99.8%, or greater than or equal to 99.9%.
- isomeric excess or “enantiomeric excess” refer to the difference between two isomers or relative percentages of two enantiomers. For example, if the content of one isomer or enantiomer is 90% and the content of the other isomer or enantiomer is 10%, the isomer or enantiomeric excess (ee value) is 80% .
- Optically active (R)- and (S)-isomers can be prepared by chiral synthesis or chiral reagents or other conventional techniques. If one enantiomer of a compound of the present invention is desired, it can be prepared by asymmetric synthesis or derivatization with a chiral auxiliary, wherein the resulting mixture of diastereomers is separated and the auxiliary group is cleaved to provide pure desired enantiomer.
- a basic functional group such as an amino group
- an acidic functional group such as a carboxyl group
- a diastereomeric salt is formed with an appropriate optically active acid or base, followed by conventional methods known in the art.
- the diastereoisomers were resolved and the pure enantiomers recovered.
- the separation of enantiomers and diastereomers is usually accomplished by the use of chromatography using a chiral stationary phase, optionally in combination with chemical derivatization (eg, from amines to amino groups) formate).
- the compounds of the present invention may contain unnatural proportions of atomic isotopes at one or more of the atoms that constitute the compound.
- compounds can be labeled with radioisotopes, such as tritium ( 3 H), iodine-125 ( 125 I) or C-14 ( 14 C).
- deuterated drugs can be formed by replacing hydrogen with deuterium, and the bond formed by deuterium and carbon is stronger than the bond formed by ordinary hydrogen and carbon. Compared with undeuterated drugs, deuterated drugs can reduce toxic side effects and increase drug stability. , enhance the efficacy, prolong the biological half-life of drugs and other advantages. All transformations of the isotopic composition of the compounds of the present invention, whether radioactive or not, are included within the scope of the present invention.
- C 1-3 alkyl is used to denote a straight or branched chain saturated hydrocarbon group consisting of 1 to 3 carbon atoms.
- the C 1-3 alkyl group includes C 1-2 and C 2-3 alkyl groups, etc.; it can be monovalent (eg methyl), divalent (eg methylene) or multivalent (eg methine) .
- Examples of C1-3 alkyl groups include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), and the like.
- S- NH2 in the compounds of the present invention is used to indicate that the carboxyl group on serine is replaced with an amide.
- the structure of the compound of the present invention can be confirmed by conventional methods well known to those skilled in the art. If the present invention relates to the absolute configuration of the compound, the absolute configuration can be confirmed by conventional technical means in the art. For example, single crystal X-ray diffraction method (SXRD), the cultured single crystal is collected by Bruker D8 venture diffractometer, the light source is CuK ⁇ radiation, and the scanning mode is: After scanning and collecting relevant data, the crystal structure was further analyzed by the direct method (Shelxs97), and the absolute configuration could be confirmed.
- SXRD single crystal X-ray diffraction method
- the cultured single crystal is collected by Bruker D8 venture diffractometer
- the light source is CuK ⁇ radiation
- the scanning mode is: After scanning and collecting relevant data, the crystal structure was further analyzed by the direct method (Shelxs97), and the absolute configuration could be confirmed.
- the compounds of the present invention can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments enumerated below, embodiments formed in combination with other chemical synthesis methods, and those well known to those skilled in the art Equivalent to alternatives, preferred embodiments include, but are not limited to, the embodiments of the present invention.
- the solvent used in the present invention is commercially available.
- Step 2 Add 20% piperidine/DMF (50 mL) to the reaction column, bubble with nitrogen for 20 minutes, and drain until no liquid flows out. DMF (50 mL) was added for 5 washes of 1 min each and drained until no liquid came out. Ninhydrin detection, resin blue.
- the dried peptide resin was added to the prepared cleavage solution, shaken for 2.5 h on a shaker, filtered, and the filtrate was added to 10 times the volume of ice isopropyl ether, centrifuged, and washed three times with isopropyl ether.
- the crude peptide was obtained by vacuum drying for 2 hours, and purified to obtain the polypeptide compound WX-001.
- the molecular weight of the polypeptide was confirmed by ESI-MS, the calculated value was 4811.4, and the detected value was 4812.0.
- the dried peptide resin was added to the prepared cleavage solution, shaken on a shaker for 2.5 h, filtered, and the filtrate was added to 10-fold volume of ice isopropyl ether, centrifuged, and washed three times with isopropyl ether.
- the crude peptide was obtained after drying in vacuum for 2 h, and purified to obtain the polypeptide compound WX-002.
- the molecular weight of the polypeptide was confirmed by ESI-MS, the calculated value was 4940.5, and the detected value was 4940.6.
- the dried peptide resin was added to the prepared cleavage solution, shaken for 2.5 h on a shaker, filtered, and the filtrate was added to 10 times the volume of ice isopropyl ether, centrifuged, and washed three times with isopropyl ether.
- the crude peptide was obtained by vacuum drying for 2 h, and purified to obtain the polypeptide compound WX-003.
- the molecular weight of the polypeptide was confirmed by ESI-MS, the calculated value was 4912.5, and the detected value was 4913.1.
- WX-001 Referring to the synthesis of WX-001, WX-004 was obtained through intermediate B-2. The molecular weight of the polypeptide was confirmed by ESI-MS, the calculated value was 4797.4, and the detected value was 4797.3.
- WX-001 Referring to the synthesis of WX-001, WX-005 was obtained through intermediate B-3.
- the molecular weight of the polypeptide was confirmed by ESI-MS, the calculated value was 4869.5, and the detected value was 4869.3.
- WX-001 Referring to the synthesis of WX-001, WX-006 was obtained through intermediate B-4. The molecular weight of the polypeptide was confirmed by ESI-MS, the calculated value was 4825.5, and the detected value was 4825.5.
- WX-001 Referring to the synthesis of WX-001, WX-007 was obtained through intermediate B-5. The molecular weight of the polypeptide was confirmed by ESI-MS, the calculated value was 4811.4, and the detected value was 4811.1.
- WX-001 Referring to the synthesis of WX-001, WX-008 was obtained through intermediate B-6. The molecular weight of the polypeptide was confirmed by ESI-MS, the calculated value was 4811.4, and the detected value was 4811.4.
- WX-001 Referring to the synthesis of WX-001, WX-009 was obtained through intermediate B-7.
- the molecular weight of the polypeptide was confirmed by ESI-MS, the calculated value was 4854.5, and the detected value was 4854.9.
- WX-010 was obtained through intermediate B-8.
- the molecular weight of the polypeptide was confirmed by ESI-MS, the calculated value was 4839.4, and the detected value was 4839.6.
- the cell line was constructed by Shanghai WuXi AppTec. See the table below for details.
- article number factory cAMP detection kit 29F 62AM4PEJ Cisbio 1M HEPES 2120919 15630-106 Invitrogen Hanks Balanced Salt Solution (HBSS) 2185775 14025 Invitrogen Human Serum Albumin (HSA) SLCF7301 A1653-10G Sigma casein SLCC9458 C4765-10mL Sigma 3-Isobutyl-1-methylxanthine (IBMX) STBF6061V I5879-5G Sigma ECHO qualified 384-well plate 0006433672 PP-0200 Labcyte OptiPlate-384 8210-19481 6007299 Perkin Elmer
- the compounds to be tested were diluted 4 times in 10 points, the initial concentration was 30 ⁇ M, and Bravo completed the dilution.
- the compounds of the present invention have strong agonistic activity on GLP-1R/GIPR.
- Candidate compounds were formulated into clear solutions and administered to rats by a single subcutaneous injection (SC, 0.048mpk).
- Whole blood was collected, plasma was prepared, drug concentration was analyzed by LC-MS/MS method, and pharmacokinetic parameters were calculated by Phoenix WinNonlin software.
- the compounds of the present invention have excellent pharmacokinetic properties in rats.
- the pharmacokinetic characteristics in rodents after intravenous injection and subcutaneous injection of compounds were tested according to the standard protocol.
- the candidate compounds were formulated into clear solutions and administered to mice by a single intravenous injection (IV, 0.048mpk) and subcutaneous injection (SC). , 0.048mpk).
- Whole blood was collected, plasma was prepared, drug concentration was analyzed by LC-MS/MS method, and pharmacokinetic parameters were calculated by Phoenix WinNonlin software.
- the compounds of the present invention have excellent pharmacokinetic properties in mice.
- the mammalian pharmacokinetic characteristics of the compounds after intravenous injection and subcutaneous injection were tested according to the standard protocol.
- the candidate compounds were formulated into clear solutions and administered to cynomolgus monkeys by a single subcutaneous injection (SC, 0.02mpk).
- Whole blood was collected, plasma was prepared, drug concentration was analyzed by LC-MS/MS method, and pharmacokinetic parameters were calculated by Phoenix WinNonlin software.
- the compounds of the present invention have excellent pharmacokinetic properties in monkeys.
- test compounds in normal mouse plasma was investigated.
- test compound solutions Dilute with DMSO to make 100 ⁇ M solutions.
- the compounds of the present invention have excellent plasma stability.
- Matrix preparation On the day of the experiment, the plasma was thawed in cold water and centrifuged at 3220 rpm for 5 min to remove all blood clots. The pH of the resulting plasma was measured and adjusted to 7.4 ⁇ 0.1 using 1% phosphoric acid or 1 N sodium hydroxide as needed.
- test compounds were dissolved in dimethyl sulfoxide (DMSO) to prepare stock solutions with concentrations of 10 mM and 2 mM, respectively.
- a 40 ⁇ M working solution was prepared by diluting 2 ⁇ L of the stock solution (2 mM) with 98 ⁇ L DMSO.
- a 400 ⁇ M working solution of the control compound was prepared by diluting 10 ⁇ L of the stock solution with 240 ⁇ L DMSO.
- the loading matrix was prepared by mixing the working solution of the compound (5 ⁇ L) with the blank matrix (995 ⁇ L) at a ratio of 1:200.
- test samples an additional aliquot of the matrix-containing sample was transferred to a separate 96-well plate (sample incubation plate) and incubated at 37°C for 4 h.
- the compounds of the present invention have a very high degree of plasma protein binding.
- test compounds in normal mouse kidney homogenate was investigated.
- Test compound dilute 10 mM stock solution with DMSO to prepare 1 mM intermediate solution;
- Test compound dilute 1 mM intermediate solution with DMSO to prepare 50 ⁇ M dosing solution;
- Experimental example 8 Compound mouse intraperitoneal glucose tolerance (ipGTT) test-in vivo efficacy evaluation
- each group of animals was injected with the test compound (0.3nmol/kg) and the vehicle (20mM citrate buffer), fasted overnight, and intraperitoneally injected with glucose solution (2g) after 18 hours. /kg, 10mL/kg);
- the compounds of the present invention have excellent glucose tolerance-improving effect.
- mice After db/db mice arrive at the facility, they are kept in an animal breeding room with strictly controlled environmental conditions. The temperature in the breeding room is maintained at 20-24°C and the humidity is maintained at 30-70%. The temperature and humidity in the breeding room were monitored in real time by a thermo-hygrometer, and the temperature and humidity were recorded twice a day (once in the morning and once in the afternoon). The lighting in the animal breeding room is controlled by an electronic timed light-on system, with the lights on for 12 hours a day and off for 12 hours (on at 7:00 am and off at 19:00 in the afternoon). During the experiment, animals were housed in single cages, and toys were provided in each cage. During the experiment, animals had free access to food (growth/reproduction feed for rats and mice) and drinking water.
- the vehicle and the test compound (15 nmol/kg) were subcutaneously injected into each group of animals, administration time: 9:30-11:00 in the morning, once a day, for 4 consecutive weeks.
- the compounds of the present invention exhibited excellent hypoglycemic efficacy in db/db mice.
- the DIO mice arrive at the WuXi AppTec facility, they are kept in an animal breeding room with strictly controlled environmental conditions.
- the temperature in the breeding room is maintained at 20-24°C and the humidity is maintained at 30-70%.
- the temperature and humidity in the breeding room were monitored in real time by a thermohygrometer, and the temperature and humidity were recorded twice a day (once in the morning and once in the afternoon).
- the lighting in the animal breeding room is controlled by an electronic timed light-on system, with the lights on for 12 hours a day and off for 12 hours (on at 7:00 am and off at 19:00 in the afternoon).
- animals were housed in single cages, and toys were provided in each cage.
- animals had free access to food (growth/reproduction feed for rats and mice) and drinking water.
- the vehicle and the test compound (10 nmol/kg) were subcutaneously injected into each group of animals, the administration time: 9:30 in the morning, the administration frequency was once every three days, and the administration period was 22 days.
Abstract
一类含内酰胺修饰的多肽类化合物，及其在制备治疗相关疾病的药物中的应用。
Description
本申请主张如下优先权：
CN202011402979.7，申请日：2020年12月02日；
CN202011409947.X，申请日：2020年12月02日；
CN202110432060.0，申请日：2021年04月21日；
CN202110587056.1，申请日：2021年05月27日。
本发明涉及一类含内酰胺修饰的多肽类化合物，及其在制备治疗相关疾病的药物中的应用。
中国预估有1.1亿糖尿病患者，占全世界糖尿病患者的24％。随着经济发展与生活方式的改变，我国糖尿病患病率增加至12.8％(部分省市高达19.9％)，并伴有肥胖或心血管疾病(CVD)。最近研究发现，葡萄糖依赖性促胰岛素样肽(GIP)/胰高血糖素样肽-1(GLP-1)双重激动剂可用于糖尿病治疗。
GLP-1在胰岛中发挥保护胰岛β细胞的作用，以葡萄糖依赖的方式刺激胰岛β细胞释放胰岛素，有效控制餐后血糖的作用。因其独特的作用机制，低血糖风险大大降低。虽然GLP-1R激动剂在临床中展现了优秀的降糖效果，但还是有很多二型糖尿病人未达到降糖及减重目标。因此，将GLP-1R激动剂与其它降糖靶点结合如GIP用于二型糖尿病治疗是一个迫切且有前景的需求。GIP是由小肠的神经内分泌K细胞分泌的多肽，由GIPR介导其生理作用，主要为非葡萄糖依赖的促胰岛素分泌、增强胰高血糖素分泌、增强脂质代谢等。虽然GIPR激动剂的有益作用似乎在2型糖尿病患者的高血糖症状中减弱，但研究表明，GIP减弱的促胰岛素分泌作用可以在血浆葡萄糖水平恢复正常一段时间后完全恢复。这表明共同激动GLP-1R/GIPR可发挥协同降血糖作用。GIP和GLP-1双重激动对糖/脂代谢的调节功能具有协同效应，具有更优的降血糖、减体重、缓解肝脏脂肪的治疗效果。
发明内容
本发明提供了下式化合物或其药学上可接受的盐，
SEQ ID NO1：YAibEGT FTSDY SIAibLD KK
1AZ
0K
0Z
1FZ
2E
0W LZ
3AGG PSSGA PPPS
0
SEQ ID NO2：YAibEGT FTSDY SIAibLD KE
0AQK
0AFVK
1W LIAGG PSSGA PPPS
0
SEQ ID NO3：YAibEGT FTSDY SIE
0LD KK
0AQK
1AFVQW LIAGG PSSGA PPPS
0
SEQ ID NO4：YAibEGT FTSDY SIE
0LD K
0IAQK
1AFVQW LIAGG PSSGA PPPS
0
其中，
Aib的结构为
S
0选自
Z
0选自谷氨酰胺(Q)和天冬酰胺(N)；
Z
1选自丙氨酸(A)和谷氨酸(E)；
Z
2选自缬氨酸(V)和异亮氨酸(I)；
Z
3选自异亮氨酸(I)和亮氨酸(L)；
E
0和K
0表示谷氨酸侧链上的羧基与赖氨酸侧链上的氨基共同构成内酰胺，此时K
0Z
1FZ
2E
0的结构为
E
0AQK
0的结构为
E
0LDKK
0的结构为
E
0LDK
0的结构为
K
1表示赖氨酸侧链上的氨基与-X-X
1-X
2相连，其结构为
X选自
R
1和R
2分别独立地选自H和CH
3；
X
1选自
X
2选自
m、n和p分别独立地选自2和3；
s选自2和3；
q选自15、16、17、18和19。
在本发明的一些方案中，上述化合物或其药学上可接受的盐，其选自：
其中，
Aib、Z
0、Z
1、Z
2、Z
3和K
1如本发明所定义。
在本发明的一些方案中，上述X选自-C(＝O)-CH
2-，其他变量如本发明所定义。
在本发明的一些方案中，上述m、n和p分别独立地选自2，其他变量如本发明所定义。
在本发明的一些方案中，上述s选自2，其他变量如本发明所定义。
在本发明的一些方案中，上述q选自15和17，其他变量如本发明所定义。
在本发明的一些方案中，上述X
1选自
其他变量如本发明所定义。
在本发明的一些方案中，上述X
2选自
其他变量如本发明所定义。
在本发明的一些方案中，上述-X-X
1-X
2选自
其他变量如本发明所定义。
本发明提供了下式化合物或其药学上可接受的盐，
YAibEGT FTSDY SIAibLD KK
1AQK
0AFVE
0W LIAGG PSSGA PPPS
0
YAibEGT FTSDY SIAibLD KK
1ANK
0AFVE
0W LIAGG PSSGA PPPS
0
YAibEGT FTSDY SIAibLD KK
1AQK
0EFVE
0W LIAGG PSSGA PPPS
0
YAibEGT FTSDY SIAibLD KK
1AQK
0AFIE
0W LIAGG PSSGA PPPS
0
YAibEGT FTSDY SIAibLD KK
1AQK
0AFVE
0W LLAGG PSSGA PPPS
0
其中，
Aib的结构为
S
0选自
K
0和E
0表示赖氨酸侧链上的氨基与谷氨酸侧链上的羧基共同构成内酰胺，其结构为
K
1表示赖氨酸侧链上的氨基与-X-X
1-X
2相连，其结构为
X选自
R
1和R
2分别独立地选自H；
X
1选自、
X
2选自
m和n分别独立地选自2和3；
s选自2和3；
q选自15、16、17、18和19。
在本发明的一些方案中，上述X选自-C(＝O)-CH
2-，其他变量如本发明所定义。
在本发明的一些方案中，上述m和n分别独立地选自2，其他变量如本发明所定义。
在本发明的一些方案中，上述s选自2，其他变量如本发明所定义。
在本发明的一些方案中，上述q选自15和17，其他变量如本发明所定义。
在本发明的一些方案中，上述X
1选自
其他变量如本发明所定义。
在本发明的一些方案中，上述X
2选自
其他变量如本发明所定义。
在本发明的一些方案中，上述-X-X
1-X
2选自
其他变量如本发明所定义。
本发明提供了下式化合物或其药学上可接受的盐，
YAibEGT FTSDY SIAibLD KK
1AZ
0K
0Z
1FZ
2E
0W LZ
3AGG PSSGA PPPS
0
YAibEGT FTSDY SIAibLD KE
0AQK
0AFVK
1W LIAGG PSSGA PPPS
0
YAibEGT FTSDY SIE
0LD KK
0AQK
1AFVQW LIAGG PSSGA PPPS
0
其中，
Aib的结构为
S
0选自
Z
0选自谷氨酰胺(Q)和天冬酰胺(N)；
Z
1选自丙氨酸(A)和谷氨酸(E)；
Z
2选自缬氨酸(V)和异亮氨酸(I)；
Z
3选自异亮氨酸(I)和亮氨酸(L)；
E
0和K
0表示谷氨酸侧链上的羧基与赖氨酸侧链上的氨基共同构成内酰胺，此时K
0Z
1FZ
2E
0的结构为
E
0AQK
0的结构为
E
0LDKK
0的结构为
K
1表示赖氨酸侧链上的氨基与-X-X
1-X
2相连，其结构为
X选自
R
1和R
2分别独立地选自H；
X
1选自
X
2选自
m、n和p分别独立地选自2和3；
s独立地选自2和3；
q独立地选自15、16、17、18和19。
在本发明的一些方案中，上述X选自-C(＝O)-CH
2-，其他变量如本发明所定义。
在本发明的一些方案中，上述m、n和p分别独立地选自2，其他变量如本发明所定义。
在本发明的一些方案中，上述s独立地选自2，其他变量如本发明所定义。
在本发明的一些方案中，上述q独立地选自15和17，其他变量如本发明所定义。
在本发明的一些方案中，上述q独立地选自17，其他变量如本发明所定义。
在本发明的一些方案中，上述X
1选自
其他变量如本发明所定义。
在本发明的一些方案中，上述X
2选自
其他变量如本发明所定义。
在本发明的一些方案中，上述-X-X
1-X
2选自
其他变量如本发明所定义。
本发明提供了下式化合物或其药学上可接受的盐，
YAibEGT FTSDY SIAibLD KK
1AQK
0AFVE
0W LIAGG PSSGA PPPS
0
其中，
Aib的结构为
S
0选自
K
0和E
0表示赖氨酸侧链上的氨基与谷氨酸侧链上的羧基共同构成内酰胺，其结构为
K
1表示赖氨酸侧链上的氨基与-X-X
1-X
2相连，其结构为
X选自
R
1和R
2分别独立地选自H；
X
1选自
X
2选自
m、n和p分别独立地选自2和3；
q独立地选自15、16、17、18和19。
在本发明的一些方案中，上述X选自-C(＝O)-CH
2-，其他变量如本发明所定义。
在本发明的一些方案中，上述m、n和p分别独立地选自2，其他变量如本发明所定义。
在本发明的一些方案中，上述q独立地选自17，其他变量如本发明所定义。
在本发明的一些方案中，上述X
1选自
其他变量如本发明所定义。
在本发明的一些方案中，上述X
2选自
其他变量如本发明所定义。
在本发明的一些方案中，上述-X-X
1-X
2选自
本发明提供了下式化合物或其药学上可接受的盐，
YAibEGT FTSDY SIE
0LD K
0IAQK
1AFVQW LIAGG PSSGA PPPS
0
其中，
Aib的结构为
S
0选自
E
0和K
0表示谷氨酸侧链上的羧基与赖氨酸侧链上的氨基共同构成内酰胺，其结构为
K
1表示赖氨酸侧链上的氨基与-X-X
1-X
2相连，其结构为
X选自
R
1和R
2分别独立地选自H；
X
1选自
X
2选自
m、n和p分别独立地选自2和3；
q独立地选自15、16、17、18和19。
在本发明的一些方案中，上述X选自-C(＝O)-CH
2-，其他变量如本发明所定义。
在本发明的一些方案中，上述m、n和p分别独立地选自2，其他变量如本发明所定义。
在本发明的一些方案中，上述q独立地选自17，其他变量如本发明所定义。
在本发明的一些方案中，上述X
1选自
其他变量如本发明所定义。
在本发明的一些方案中，上述X
2选自
其他变量如本发明所定义。
在本发明的一些方案中，上述-X-X
1-X
2选自
其他变量如本发明所定义。
本发明还有一些方案由上述变量任意组合而来。
本发明还提供了下式所示化合物或其药学上可接受的盐，
本发明还提供了下式所示化合物或其药学上可接受的盐，
本发明的一些方案中，上述化合物或其药学上可接受的盐在制备治疗肥胖症及糖尿病的药物上的应用。
本发明还提供了下述测试方法：
实验例：hERG钾离子通道的抑制试验
A.主要材料
CHO-hERG细胞系(稳定表达hERG通道的中国仓鼠卵巢细胞)，中国科学院上海药物研究所内部构建；膜片钳放大器Axopatch 200B,泰瑟国际公司；
B.方法
a)细胞培养
稳定表达hERG的CHO细胞培养于直径35mm的细胞培养皿中，置于37℃，5％CO
2的培养箱培养，每48小时按1:5比例进行传代，培养基配方：90％F12(Invitrogen)，10％胎牛血清(Gibco)，100μg/mL G418(Invitrogen)和100μg/mL Hygromycin B(Invitrogen)。试验当天，吸走细胞培养液，用细胞外液淋洗一遍后加入0.25％Trypsin-EDTA(Invitrogen)溶液，在室温下消化3～5分钟。吸走消化液，用细胞外液重悬后将细胞转移到用于电生理记录的实验皿中备用。
b)细胞内外液的配制
细胞外液需每个月配制一次。细胞内液须分装冻存在-20℃。
细胞外液(mM)：145NaCl,4KCl,2CaCl
2,1MgCl
2,10Glucose和10HEPES，用NaOH调节pH至7.4，渗透压295mOsm。
细胞内液(mM)：120KCl,31.25KOH,5.374CaCl
2,1.75MgCl
2,4Na
2ATP,10HEPES和10EGTA，用KOH 调节pH至7.2，渗透压285mOsm。
c)化合物准备
化合物用DMSO溶解成20mM母液，测试当天，将化合物母液用DMSO进行3倍连续稀释，即取10μL的化合物母液加入到20μL DMSO中，依次得到6个经DMSO连续稀释的化合物中间浓度，依次分别为20、6.66、2.22、0.74、0.24和0.082mM。然后再取10μL的化合物中间浓度加入到4990μL细胞外液中，500倍稀释得到需要测试的最终浓度，最高测试浓度为40μM，依次分别为40、13.3、4.44、1.48、0.49和0.16μM。最终测试浓度中DMSO的含量不超过0.2％，此浓度的DMSO对hERG钾通道没有影响。
d)电生理记录过程
稳定表达hERG钾通道的CHO(Chinese Hamster Ovary)细胞，在室温下用全细胞膜片钳技术记录hERG钾通道电流。玻璃微电极由玻璃电极毛胚(BF150-86-10，Sutter)经拉制仪拉制而成，灌注电极内液后的尖端电阻为2-5MΩ左右，将玻璃微电极插入放大器探头即可连接至Axopatch 200B(Molecular Devices)膜片钳放大器。钳制电压和数据记录由pClamp 10软件通过电脑控制和记录，采样频率为10kHz，滤波频率为2kHz。在得到全细胞记录后，细胞钳制在-80mV，诱发hERG钾电流(IhERG)的步阶电压从-80mV给予一个2s的去极化电压到+20mV，再复极化到-50mV，持续1s后回到-80mV。每10s给予此电压刺激，确定hERG钾电流稳定后(1分钟)开始给药过程。化合物浓度从低测试浓度开始连续给药，每个测试浓度至少给予1分钟。化合物每个浓度至少测试3个细胞(n≥3)，阳性化合物每个浓度至少测试2个细胞(n≥2)。
e)数据分析
在每一个完整电流记录中，基于峰值电流在阴性对照中所占的百分比，可以计算出每一化合物作用浓度的抑制百分比。利用标准希式方程拟合得到量效关系曲线，具体方程如下：
I
(C)＝I
b+(I
fr-I
b)*c
n/(IC
50
n+c
n)
c为化合物测试浓度，n为斜率
曲线拟合和抑制率计算均由Qpatch分析软件分析完成，若最低浓度下抑制率超过半数抑制或最高浓度下抑制率未达到半数抑制，则该化合物相应的IC
50低于最低浓度或IC
50值大于最高浓度。
C.测试结果和结论
本发明化合物无hERG相关风险。
实验例：细胞色素P450同工酶抑制性
A.实验目的
测定受试化合物对人肝微粒体细胞色素P450同工酶(CYP1A2、CYP2C9、CYP2C19、CYP2D6和CYP3A4)活性的抑制作用。
B.实验操作
首先将受试化合物(10mM)进行梯度稀释，制备工作液(100×最终浓度)，工作液浓度分别为：5,1.5,0.5,0.15,0.05,0.015,0.005mM，同时准备P450同工酶(CYP1A2、CYP2C9、CYP2C19、CYP2D6和CYP3A4)各阳性抑制剂及其特异性底物混合物的工作液；将冷冻于–80℃冰箱的人肝微粒体置于冰上解冻，待人肝微粒体全部溶解，用PBS(磷酸缓冲液)进行稀释，制备一定浓度工作液(0.253mg/ml)；并将20μL底物混合液加至反应板中(Blank孔中加入20μL PB)同时将158μL人肝微粒体工作液加入反应板中，将反应板置于冰上，待用；此时将2μL各个浓度的受试化合物(N＝1)及阳性抑制剂(N＝2)加入对应孔中，无抑制剂(受试化合物或阳性抑制剂)组加入对应的有机溶剂，作为对照组样品；在37℃水浴预孵育10min后，将20μL NADPH溶液加入反应板中，置于37℃水浴孵育10min；加入400μL冷的乙腈溶液终止反应；将反应板置于摇床，振荡10min；4,000rpm离心20min；取200μL上清加至100μL水中，进行样品稀释；最后封板，振荡，摇匀，进行LC/MS/MS检测。
C.实验结果和结论
本发明化合物无相关CYP抑制风险。
技术效果
本发明化合物对GLP-1R/GIPR具有很强的激动活性；本发明化合物具有优异的药代动力学性质；本发明化合物具有优异的血浆稳定性；本发明化合物具有极高的血浆蛋白结合度；本发明化合物具有优异的体内药效。
定义和说明
除非另有说明，本文所用的下列术语和短语旨在具有下列含义。一个特定的术语或短语在没有特别定义的情况下不应该被认为是不确定的或不清楚的，而应该按照普通的含义去理解。当本文中出现商品名时，意在指代其对应的商品或其活性成分。
这里所采用的术语“药学上可接受的”，是针对那些化合物、材料、组合物和/或剂型而言，它们在可靠的医学判断的范围之内，适用于与人类和动物的组织接触使用，而没有过多的毒性、刺激性、过敏性反应或其它问题或并发症，与合理的利益/风险比相称。
术语“药学上可接受的盐”是指本发明化合物的盐，由本发明发现的具有特定取代基的化合物与相对无毒的酸或碱制备。当本发明的化合物中含有相对酸性的功能团时，可以通过在纯的溶液或合适的惰性溶剂中用足够量的碱与这类化合物接触的方式获得碱加成盐。药学上可接受的碱加成盐包括钠、钾、钙、铵、 有机胺或镁盐或类似的盐。当本发明的化合物中含有相对碱性的官能团时，可以通过在纯的溶液或合适的惰性溶剂中用足够量的酸与这类化合物接触的方式获得酸加成盐。药学上可接受的酸加成盐的实例包括无机酸盐，所述无机酸包括例如盐酸、氢溴酸、硝酸、碳酸，碳酸氢根，磷酸、磷酸一氢根、磷酸二氢根、硫酸、硫酸氢根、氢碘酸、亚磷酸等；以及有机酸盐，所述有机酸包括如乙酸、丙酸、异丁酸、马来酸、丙二酸、苯甲酸、琥珀酸、辛二酸、反丁烯二酸、乳酸、扁桃酸、邻苯二甲酸、苯磺酸、对甲苯磺酸、柠檬酸、酒石酸和甲磺酸等类似的酸；还包括氨基酸(如精氨酸等)的盐，以及如葡糖醛酸等有机酸的盐。本发明的某些特定的化合物含有碱性和酸性的官能团，从而可以被转换成任一碱或酸加成盐。
本发明的药学上可接受的盐可由含有酸根或碱基的母体化合物通过常规化学方法合成。一般情况下，这样的盐的制备方法是：在水或有机溶剂或两者的混合物中，经由游离酸或碱形式的这些化合物与化学计量的适当的碱或酸反应来制备。
本文所述的A或Ala表示丙氨酸，结构为
R或Arg表示精氨酸，结构为
N或Asn表示天冬酰胺，结构为
D或Asp表示天冬氨酸，结构为
C或Cys表示半胱氨酸，结构为
Q或Gln表示谷氨酰胺，结构为
E或Glu表示谷氨酸，结构为
G或Gly表示甘氨酸，结构为
H或His表示组氨酸，结构为
I或Ile表示异亮氨酸，结构为
L或Leu表示亮氨酸，结构为
K或Lys表示赖氨酸，结构为
M或Met表示甲硫氨酸，结构为
F或Phe表示苯丙氨 酸，结构为
P或Pro表示脯氨酸，结构为
S或Ser表示丝氨酸，结构为
T或Thr表示苏氨酸，结构为
W或Trp表示色氨酸，结构为
Y或Tyr表示酪氨酸，结构为
V或Val表示缬氨酸，结构为
术语“治疗”包括抑制、减缓、停止或逆转现有症状或病患的进展或严重程度。
除非另有说明，术语“异构体”意在包括几何异构体、顺反异构体、立体异构体、对映异构体、旋光异构体、非对映异构体和互变异构体。
本发明的化合物可以存在特定的几何或立体异构体形式。本发明设想所有的这类化合物，包括顺式和反式异构体、(-)-和(+)-对映体、(R)-和(S)-对映体、非对映异构体、(D)-异构体、(L)-异构体，及其外消旋混合物和其他混合物，例如对映异构体或非对映体富集的混合物，所有这些混合物都属于本发明的范围之内。烷基等取代基中可存在另外的不对称碳原子。所有这些异构体以及它们的混合物，均包括在本发明的范围之内。
除非另有说明，术语“对映异构体”或者“旋光异构体”是指互为镜像关系的立体异构体。
除非另有说明，术语“顺反异构体”或者“几何异构体”系由因双键或者成环碳原子单键不能自由旋转而引起。
除非另有说明，术语“非对映异构体”是指分子具有两个或多个手性中心，并且分子间为非镜像的关系的立体异构体。
除非另有说明，“(+)”表示右旋，“(-)”表示左旋，“(±)”表示外消旋。
除非另有说明，用楔形实线键
和楔形虚线键
表示一个立体中心的绝对构型，用直形实线键
和直形虚线键
表示立体中心的相对构型，用波浪线
表示楔形实线键
或楔形虚线键
或用波浪线
表示直形实线键
或直形虚线键
除非另有说明，术语“富含一种异构体”、“异构体富集”、“富含一种对映体”或者“对映体富集”指其中一 种异构体或对映体的含量小于100％，并且，该异构体或对映体的含量大于等于60％，或者大于等于70％，或者大于等于80％，或者大于等于90％，或者大于等于95％，或者大于等于96％，或者大于等于97％，或者大于等于98％，或者大于等于99％，或者大于等于99.5％，或者大于等于99.6％，或者大于等于99.7％，或者大于等于99.8％，或者大于等于99.9％。
除非另有说明，术语“异构体过量”或“对映体过量”指两种异构体或两种对映体相对百分数之间的差值。例如，其中一种异构体或对映体的含量为90％，另一种异构体或对映体的含量为10％，则异构体或对映体过量(ee值)为80％。
可以通过的手性合成或手性试剂或者其他常规技术制备光学活性的(R)-和(S)-异构体以及D和L异构体。如果想得到本发明某化合物的一种对映体，可以通过不对称合成或者具有手性助剂的衍生作用来制备，其中将所得非对映体混合物分离，并且辅助基团裂开以提供纯的所需对映异构体。或者，当分子中含有碱性官能团(如氨基)或酸性官能团(如羧基)时，与适当的光学活性的酸或碱形成非对映异构体的盐，然后通过本领域所公知的常规方法进行非对映异构体拆分，然后回收得到纯的对映体。此外，对映异构体和非对映异构体的分离通常是通过使用色谱法完成的，所述色谱法采用手性固定相，并任选地与化学衍生法相结合(例如由胺生成氨基甲酸盐)。
本发明的化合物可以在一个或多个构成该化合物的原子上包含非天然比例的原子同位素。例如，可用放射性同位素标记化合物，比如氚(
3H)，碘-125(
125I)或C-14(
14C)。又例如，可用重氢取代氢形成氘代药物，氘与碳构成的键比普通氢与碳构成的键更坚固，相比于未氘化药物，氘代药物有降低毒副作用、增加药物稳定性、增强疗效、延长药物生物半衰期等优势。本发明的化合物的所有同位素组成的变换，无论放射性与否，都包括在本发明的范围之内。
除非另有规定，术语“C
1-3烷基”用于表示直链或支链的由1至3个碳原子组成的饱和碳氢基团。所述C
1-3烷基包括C
1-2和C
2-3烷基等；其可以是一价(如甲基)、二价(如亚甲基)或者多价(如次甲基)。C
1-
3烷基的实例包括但不限于甲基(Me)、乙基(Et)、丙基(包括n-丙基和异丙基)等。
除非另有规定，本发明化合物中“S-NH
2”用于表示丝氨酸上的羧基替换为酰胺。
本发明的化合物可以通过本领域技术人员所熟知的常规方法来确认结构，如果本发明涉及化合物的绝对构型，则该绝对构型可以通过本领域常规技术手段予以确证。例如单晶X射线衍射法(SXRD)，把培养出的单晶用Bruker D8 venture衍射仪收集衍射强度数据，光源为CuKα辐射，扫描方式：
扫描，收集相关数据后，进一步采用直接法(Shelxs97)解析晶体结构，便可以确证绝对构型。
本发明的化合物可以通过本领域技术人员所熟知的多种合成方法来制备，包括下面列举的具体实施方式、其与其他化学合成方法的结合所形成的实施方式以及本领域技术上人员所熟知的等同替换方式，优选的实施方式包括但不限于本发明的实施例。
本发明所使用的溶剂可经市售获得。
本发明采用下述缩略词：aq代表水；eq代表当量、等量；DCM代表二氯甲烷；PE代表石油醚；DMSO代表二甲亚砜；MeOH代表甲醇；BOC代表叔丁氧羰基是一种胺保护基团；r.t.代表室温；O/N代表过夜；THF代表四氢呋喃；Boc
2O代表二叔丁基二碳酸酯；TFA代表三氟乙酸；DIEA代表二异丙基乙基胺；DMF代表N,N-二甲基甲酰胺；HBTU代表苯并三氮唑-N,N,N',N'-四甲基脲六氟磷酸酯；HOBT代表1-羟基苯并三唑；HOAT代表1-羟基-7-氮杂苯并三唑；DIC代表N,N'-二异丙基碳二亚胺；DBU代表1,8-二氮杂双环[5.4.0]十一碳-7-烯；PhSiH
3代表苯硅烷；Pd(PPh
3)
4代表四三苯基膦钯。
下面通过实施例对本发明进行详细描述，但并不意味着对本发明任何不利限制。本文已经详细地描述了本发明，其中也公开了其具体实施例方式，对本领域的技术人员而言，在不脱离本发明精神和范围的情况下针对本发明具体实施方式进行各种变化和改进将是显而易见的。
中间体B-1
步骤1：称取1.1g 4-(2’,4’-二甲氧基苯基-芴甲氧羰基-氨甲基)-苯氧基乙酰氨基-甲基二苯甲胺树脂(替代度Sub＝0.28mmol/g)加入到反应柱中，再加入DMF(50mL)至反应柱氮气鼓气体2小时，排废，直至没有液体流出，加入DMF(50mL)洗涤5次每次1分钟，排废，直至没有液体流出。
步骤2：加入20％的哌啶/DMF(50mL)至反应柱中，氮气鼓起20分钟，排废，直至没有液体流出。加入DMF(50mL)洗涤5次每次1分钟，排废，直至没有液体流出。茚三酮检测，树脂蓝色。
步骤3：氨基酸的偶联
3.1 Fmoc-Ser(tBu)-OH的偶联
1.称取Fmoc-Ser(tBu)-OH(3.0eq)加入到上述树脂中，加入DIEA(6.00eq)补加10mL DMF至反应柱中，鼓氮气，待氨基酸溶解后加入HBTU(2.85eq)。调节好氮气使树脂均匀鼓起。
2.在25℃的环境中反应0.5h，茚三酮检测，树脂无色透明。
3.抽掉反应液，用DMF洗涤5次(50mL每次)，每次1min，排废，直至没有液体流出。
3.2 Fmoc-Pro-OH的偶联
1.加入20％的哌啶/DMF(50mL)至反应柱中，氮气鼓起20min，排废，直至没有液体流出。加入DMF(50mL)洗涤5次，每次1min，排废，直至没有液体流出。茚三酮检测，树脂蓝色。
2.称取Fmoc-Pro-OH(3.0eq)加入到上述树脂中，加入DIEA(6.00eq)补加10mL DMF至反应柱中，鼓氮气，待氨基酸溶解后加入HBTU(2.85eq)。调节好氮气使树脂均匀鼓起。
3.在25℃的环境中反应0.5h，茚三酮检测，树脂无色透明。
4.抽掉反应液，用DMF洗涤5次(50mL每次)，每次1min，排废，直至没有液体流出。
重复3.2步骤，完成以下氨基酸的偶联
| 序号 | 原料 | 偶联试剂 |
| 3.3 | Fmoc-Pro-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 3.4 | Fmoc-Pro-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 3.5 | Fmoc-Ala-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 3.6 | Fmoc-Gly-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 3.7 | Fmoc-Ser(tBu)-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 3.8 | Fmoc-Ser(tBu)-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 3.9 | Fmoc-Pro-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 3.10 | Fmoc-Gly-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 3.11 | Fmoc-Gly-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 3.12 | Fmoc-Ala-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 3.13 | Fmoc-Ile-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 3.14 | Fmoc-Leu-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 3.15 | Fmoc-Trp(Boc)-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 3.16 | Fmoc-Glu(OAll)-OH(2.00eq) | HBTU(1.90eq)and DIEA(4.00eq) |
| 3.17 | Fmoc-Val-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 3.18 | Fmoc-Phe-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 3.19 | Fmoc-Ala-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 3.20 | Fmoc-Lys(Alloc)-OH(2.00eq) | HOBT(2.00eq)and DIC(2.00eq) |
| 3.21 | Fmoc-Gln(Trt)-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 3.22 | Fmoc-Ala-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 3.23 | Fmoc-Lys(Dde)-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 3.24 | Fmoc-Lys(Boc)-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 3.25 | Fmoc-Asp(tBu)-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 3.26 | Fmoc-Leu-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 3.27 | Fmoc-Aib-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 3.28 | Fmoc-Ile-OH(6.00eq) | HOAT(6.00eq)and DIC(6.00eq) |
| 3.29 | Fmoc-Ser(tBu)-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 3.30 | Fmoc-Tyr(tBu)-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 3.31 | Fmoc-Asp(tBu)-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 3.32 | Fmoc-Ser(tBu)-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 3.33 | Fmoc-Thr(tBu)-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 3.34 | Fmoc-Phe-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 3.35 | Fmoc-Thr(tBu)-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 3.36 | Fmoc-Gly-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 3.37 | Fmoc-Glu(tBu)-OH(6.00eq) | HOBT(6.00eq)and DIC(6.00eq) |
| 3.38 | Fmoc-Aib-OH(3.00eq) | HATU(2.85eq)and DIEA(6.00eq) |
| 3.39 | Boc-Tyr(tBu)-OH(3.00eq) | HATU(2.85eq)and DIEA(6.00eq) |
3.40脱Alloc和OAll
1.加入PhSiH
3(20.0eq)和DCM(10mL)至反应柱中，氮气鼓起后加入Pd(PPh
3)
4(0.2eq)，氮气鼓起20min，反应二次，排废，直至没有液体流出。
2.用DMF洗涤5次(50mL每次)，每次1min，排废，直至没有液体流出。
3.41酰胺关环
1.加入DIEA(3.00eq)补加10mL DMF至反应柱中，鼓氮气，待溶解后加入HATU(1.5eq)。调节好氮气使树脂均匀鼓起。
2.在25℃的环境中反应0.5h，茚三酮检测，树脂无色透明。
3.用DMF洗涤5次(50mL每次)，每次1min，排废，直至没有液体流出。
3.42脱Dde
1.加入3％的水合肼/DMF(50mL)至反应柱中，氮气鼓起15min，排废，直至没有液体流出。加入DMF(50mL)洗涤5次,每次1min，排废，直至没有液体流出,得到中间体B-1。茚三酮检测，树脂蓝色。
中间体B-2
参考B-1的合成方式，完成以下氨基酸的偶联，脱Alloc和OAll，酰胺关环及脱Dde保护等操作得到B-2.
| 序号 | 原料 | 偶联试剂 |
| 1 | Fmoc-Ser(tBu)-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 2 | Fmoc-Pro-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 3 | Fmoc-Pro-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 4 | Fmoc-Pro-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 5 | Fmoc-Ala-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 6 | Fmoc-Gly-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 7 | Fmoc-Ser(tBu)-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 8 | Fmoc-Ser(tBu)-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 9 | Fmoc-Pro-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 10 | Fmoc-Gly-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 11 | Fmoc-Gly-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 12 | Fmoc-Ala-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 13 | Fmoc-Ile-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 14 | Fmoc-Leu-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 15 | Fmoc-Trp(Boc)-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 16 | Fmoc-Glu(OAll)-OH(2.00eq) | HBTU(1.90eq)and DIEA(4.00eq) |
| 17 | Fmoc-Val-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 18 | Fmoc-Phe-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 19 | Fmoc-Ala-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 20 | Fmoc-Lys(Alloc)-OH(2.00eq) | HOBT(2.00eq)and DIC(2.00eq) |
| 21 | Fmoc-Asn(Trt)-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 22 | Fmoc-Ala-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 23 | Fmoc-Lys(Dde)-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 24 | Fmoc-Lys(Boc)-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 25 | Fmoc-Asp(tBu)-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 26 | Fmoc-Leu-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 27 | Fmoc-Aib-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 28 | Fmoc-Ile-OH(6.00eq) | HOAT(6.00eq)and DIC(6.00eq) |
| 29 | Fmoc-Ser(tBu)-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 30 | Fmoc-Tyr(tBu)-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 31 | Fmoc-Asp(tBu)-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 32 | Fmoc-Ser(tBu)-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 33 | Fmoc-Thr(tBu)-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 34 | Fmoc-Phe-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 35 | Fmoc-Thr(tBu)-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 36 | Fmoc-Gly-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 37 | Fmoc-Glu(tBu)-OH(6.00eq) | HOBT(6.00eq)and DIC(6.00eq) |
| 38 | Fmoc-Aib-OH(3.00eq) | HATU(2.85eq)and DIEA(6.00eq) |
| 39 | Boc-Tyr(tBu)-OH(3.00eq) | HATU(2.85eq)and DIEA(6.00eq) |
中间体B-3
参考B-1的合成方式，完成以下氨基酸的偶联，脱Alloc和OAll，酰胺关环及脱Dde保护等操作得到B-3.
| 序号 | 原料 | 偶联试剂 |
| 1 | Fmoc-Ser(tBu)-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 2 | Fmoc-Pro-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 3 | Fmoc-Pro-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 4 | Fmoc-Pro-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 5 | Fmoc-Ala-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 6 | Fmoc-Gly-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 7 | Fmoc-Ser(tBu)-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 8 | Fmoc-Ser(tBu)-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 9 | Fmoc-Pro-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 10 | Fmoc-Gly-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 11 | Fmoc-Gly-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 12 | Fmoc-Ala-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 13 | Fmoc-Ile-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 14 | Fmoc-Leu-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 15 | Fmoc-Trp(Boc)-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 16 | Fmoc-Glu(OAll)-OH(2.00eq) | HBTU(1.90eq)and DIEA(4.00eq) |
| 17 | Fmoc-Val-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 18 | Fmoc-Phe-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 19 | Fmoc-Glu(tBu)-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 20 | Fmoc-Lys(Alloc)-OH(2.00eq) | HOBT(2.00eq)and DIC(2.00eq) |
| 21 | Fmoc-Gln(Trt)-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 22 | Fmoc-Ala-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 23 | Fmoc-Lys(Dde)-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 24 | Fmoc-Lys(Boc)-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 25 | Fmoc-Asp(tBu)-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 26 | Fmoc-Leu-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 27 | Fmoc-Aib-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 28 | Fmoc-Ile-OH(6.00eq) | HOAT(6.00eq)and DIC(6.00eq) |
| 29 | Fmoc-Ser(tBu)-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 30 | Fmoc-Tyr(tBu)-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 31 | Fmoc-Asp(tBu)-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 32 | Fmoc-Ser(tBu)-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 33 | Fmoc-Thr(tBu)-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 34 | Fmoc-Phe-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 35 | Fmoc-Thr(tBu)-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 36 | Fmoc-Gly-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 37 | Fmoc-Glu(tBu)-OH(6.00eq) | HOBT(6.00eq)and DIC(6.00eq) |
| 38 | Fmoc-Aib-OH(3.00eq) | HATU(2.85eq)and DIEA(6.00eq) |
| 39 | Boc-Tyr(tBu)-OH(3.00eq) | HATU(2.85eq)and DIEA(6.00eq) |
中间体B-4
参考B-1的合成方式，完成以下氨基酸的偶联，脱Alloc和OAll，酰胺关环及脱Dde保护等操作得到B-4.
| 序号 | 原料 | 偶联试剂 |
| 1 | Fmoc-Ser(tBu)-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 2 | Fmoc-Pro-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 3 | Fmoc-Pro-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 4 | Fmoc-Pro-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 5 | Fmoc-Ala-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 6 | Fmoc-Gly-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 7 | Fmoc-Ser(tBu)-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 8 | Fmoc-Ser(tBu)-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 9 | Fmoc-Pro-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 10 | Fmoc-Gly-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 11 | Fmoc-Gly-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 12 | Fmoc-Ala-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 13 | Fmoc-Ile-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 14 | Fmoc-Leu-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 15 | Fmoc-Trp(Boc)-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 16 | Fmoc-Glu(OAll)-OH(2.00eq) | HBTU(1.90eq)and DIEA(4.00eq) |
| 17 | Fmoc-Ile-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 18 | Fmoc-Phe-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 19 | Fmoc-Ala-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 20 | Fmoc-Lys(Alloc)-OH(2.00eq) | HOBT(2.00eq)and DIC(2.00eq) |
| 21 | Fmoc-Gln(Trt)-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 22 | Fmoc-Ala-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 23 | Fmoc-Lys(Dde)-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 24 | Fmoc-Lys(Boc)-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 25 | Fmoc-Asp(tBu)-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 26 | Fmoc-Leu-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 27 | Fmoc-Aib-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 28 | Fmoc-Ile-OH(6.00eq) | HOAT(6.00eq)and DIC(6.00eq) |
| 29 | Fmoc-Ser(tBu)-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 30 | Fmoc-Tyr(tBu)-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 31 | Fmoc-Asp(tBu)-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 32 | Fmoc-Ser(tBu)-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 33 | Fmoc-Thr(tBu)-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 34 | Fmoc-Phe-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 35 | Fmoc-Thr(tBu)-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 36 | Fmoc-Gly-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 37 | Fmoc-Glu(tBu)-OH(6.00eq) | HOBT(6.00eq)and DIC(6.00eq) |
| 38 | Fmoc-Aib-OH(3.00eq) | HATU(2.85eq)and DIEA(6.00eq) |
| 39 | Boc-Tyr(tBu)-OH(3.00eq) | HATU(2.85eq)and DIEA(6.00eq) |
中间体B-5
参考B-1的合成方式，完成以下氨基酸的偶联，脱Alloc和OAll，酰胺关环及脱Dde保护等操作得到B-5.
| 序号 | 原料 | 偶联试剂 |
| 1 | Fmoc-Ser(tBu)-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 2 | Fmoc-Pro-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 3 | Fmoc-Pro-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 4 | Fmoc-Pro-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 5 | Fmoc-Ala-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 6 | Fmoc-Gly-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 7 | Fmoc-Ser(tBu)-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 8 | Fmoc-Ser(tBu)-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 9 | Fmoc-Pro-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 10 | Fmoc-Gly-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 11 | Fmoc-Gly-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 12 | Fmoc-Ala-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 13 | Fmoc-Leu-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 14 | Fmoc-Leu-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 15 | Fmoc-Trp(Boc)-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 16 | Fmoc-Glu(OAll)-OH(2.00eq) | HBTU(1.90eq)and DIEA(4.00eq) |
| 17 | Fmoc-Val-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 18 | Fmoc-Phe-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 19 | Fmoc-Ala-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 20 | Fmoc-Lys(Alloc)-OH(2.00eq) | HOBT(2.00eq)and DIC(2.00eq) |
| 21 | Fmoc-Gln(Trt)-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 22 | Fmoc-Ala-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 23 | Fmoc-Lys(Dde)-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 24 | Fmoc-Lys(Boc)-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 25 | Fmoc-Asp(tBu)-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 26 | Fmoc-Leu-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 27 | Fmoc-Aib-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 28 | Fmoc-Ile-OH(6.00eq) | HOAT(6.00eq)and DIC(6.00eq) |
| 29 | Fmoc-Ser(tBu)-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 30 | Fmoc-Tyr(tBu)-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 31 | Fmoc-Asp(tBu)-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 32 | Fmoc-Ser(tBu)-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 33 | Fmoc-Thr(tBu)-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 34 | Fmoc-Phe-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 35 | Fmoc-Thr(tBu)-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 36 | Fmoc-Gly-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 37 | Fmoc-Glu(tBu)-OH(6.00eq) | HOBT(6.00eq)and DIC(6.00eq) |
| 38 | Fmoc-Aib-OH(3.00eq) | HATU(2.85eq)and DIEA(6.00eq) |
| 39 | Boc-Tyr(tBu)-OH(3.00eq) | HATU(2.85eq)and DIEA(6.00eq) |
中间体B-6
参考B-1的合成方式，完成以下氨基酸的偶联，脱Alloc和OAll，酰胺关环及脱Dde保护等操作得到B-6.
| 序号 | 原料 | 偶联试剂 |
| 1 | Fmoc-Ser(tBu)-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 2 | Fmoc-Pro-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 3 | Fmoc-Pro-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 4 | Fmoc-Pro-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 5 | Fmoc-Ala-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 6 | Fmoc-Gly-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 7 | Fmoc-Ser(tBu)-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 8 | Fmoc-Ser(tBu)-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 9 | Fmoc-Pro-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 10 | Fmoc-Gly-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 11 | Fmoc-Gly-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 12 | Fmoc-Ala-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 13 | Fmoc-Ile-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 14 | Fmoc-Leu-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 15 | Fmoc-Trp(Boc)-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 16 | Fmoc-Lys(Dde)-OH(3.00eq) | HBTU(1.90eq)and DIEA(4.00eq) |
| 17 | Fmoc-Val-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 18 | Fmoc-Phe-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 19 | Fmoc-Ala-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 20 | Fmoc-Lys(Alloc)-OH(2.00eq) | HOBT(2.00eq)and DIC(2.00eq) |
| 21 | Fmoc-Gln(Trt)-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 22 | Fmoc-Ala-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 23 | Fmoc-Glu(OAll)-OH(2.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 24 | Fmoc-Lys(Boc)-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 25 | Fmoc-Asp(tBu)-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 26 | Fmoc-Leu-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 27 | Fmoc-Aib-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 28 | Fmoc-Ile-OH(6.00eq) | HOAT(6.00eq)and DIC(6.00eq) |
| 29 | Fmoc-Ser(tBu)-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 30 | Fmoc-Tyr(tBu)-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 31 | Fmoc-Asp(tBu)-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 32 | Fmoc-Ser(tBu)-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 33 | Fmoc-Thr(tBu)-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 34 | Fmoc-Phe-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 35 | Fmoc-Thr(tBu)-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 36 | Fmoc-Gly-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 37 | Fmoc-Glu(tBu)-OH(6.00eq) | HOBT(6.00eq)and DIC(6.00eq) |
| 38 | Fmoc-Aib-OH(3.00eq) | HATU(2.85eq)and DIEA(6.00eq) |
| 39 | Boc-Tyr(tBu)-OH(3.00eq) | HATU(2.85eq)and DIEA(6.00eq) |
中间体B-7
参考B-1的合成方式，完成以下氨基酸的偶联，脱Alloc和OAll，酰胺关环及脱Dde保护等操作得到B-7.
| 序号 | 原料 | 偶联试剂 |
| 1 | Fmoc-Ser(tBu)-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 2 | Fmoc-Pro-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 3 | Fmoc-Pro-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 4 | Fmoc-Pro-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 5 | Fmoc-Ala-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 6 | Fmoc-Gly-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 7 | Fmoc-Ser(tBu)-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 8 | Fmoc-Ser(tBu)-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 9 | Fmoc-Pro-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 10 | Fmoc-Gly-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 11 | Fmoc-Gly-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 12 | Fmoc-Ala-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 13 | Fmoc-Ile-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 14 | Fmoc-Leu-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 15 | Fmoc-Trp(Boc)-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 16 | Fmoc-Gln(Trt)-OH(2.00eq) | HBTU(1.90eq)and DIEA(4.00eq) |
| 17 | Fmoc-Val-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 18 | Fmoc-Phe-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 19 | Fmoc-Ala-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 20 | Fmoc-Lys(Dde)-OH(3.00eq) | HOBT(2.00eq)and DIC(2.00eq) |
| 21 | Fmoc-Gln(Trt)-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 22 | Fmoc-Ala-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 23 | Fmoc-Lys(Alloc)-OH(2.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 24 | Fmoc-Lys(Boc)-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 25 | Fmoc-Asp(tBu)-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 26 | Fmoc-Leu-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 27 | Fmoc-Glu(OAll)-OH(2.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 28 | Fmoc-Ile-OH(6.00eq) | HOAT(6.00eq)and DIC(6.00eq) |
| 29 | Fmoc-Ser(tBu)-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 30 | Fmoc-Tyr(tBu)-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 31 | Fmoc-Asp(tBu)-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 32 | Fmoc-Ser(tBu)-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 33 | Fmoc-Thr(tBu)-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 34 | Fmoc-Phe-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 35 | Fmoc-Thr(tBu)-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 36 | Fmoc-Gly-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 37 | Fmoc-Glu(tBu)-OH(6.00eq) | HOBT(6.00eq)and DIC(6.00eq) |
| 38 | Fmoc-Aib-OH(3.00eq) | HATU(2.85eq)and DIEA(6.00eq) |
| 39 | Boc-Tyr(tBu)-OH(3.00eq) | HATU(2.85eq)and DIEA(6.00eq) |
中间体B-8
参考B-1的合成方式，完成以下氨基酸的偶联，脱Alloc和OAll，酰胺关环及脱Dde保护等操作得到B-8.
| 序号 | 原料 | 偶联试剂 |
| 1 | Fmoc-Ser(tBu)-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 2 | Fmoc-Pro-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 3 | Fmoc-Pro-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 4 | Fmoc-Pro-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 5 | Fmoc-Ala-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 6 | Fmoc-Gly-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 7 | Fmoc-Ser(tBu)-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 8 | Fmoc-Ser(tBu)-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 9 | Fmoc-Pro-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 10 | Fmoc-Gly-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 11 | Fmoc-Gly-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 12 | Fmoc-Ala-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 13 | Fmoc-Ile-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 14 | Fmoc-Leu-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 15 | Fmoc-Trp(Boc)-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 16 | Fmoc-Gln(Trt)-OH(2.00eq) | HBTU(1.90eq)and DIEA(4.00eq) |
| 17 | Fmoc-Val-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 18 | Fmoc-Phe-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 19 | Fmoc-Ala-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 20 | Fmoc-Lys(Dde)-OH(3.00eq) | HOBT(2.00eq)and DIC(2.00eq) |
| 21 | Fmoc-Gln(Trt)-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 22 | Fmoc-Ala-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 23 | Fmoc-Ile-OH(3.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 24 | Fmoc-Lys(Alloc)-OH(2.00eq) | HBTU(2.85eq)and DIEA(6.00eq) |
| 25 | Fmoc-Asp(tBu)-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 26 | Fmoc-Leu-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 27 | Fmoc-Glu(OAll)-OH(2.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 28 | Fmoc-Ile-OH(6.00eq) | HOAT(6.00eq)and DIC(6.00eq) |
| 29 | Fmoc-Ser(tBu)-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 30 | Fmoc-Tyr(tBu)-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 31 | Fmoc-Asp(tBu)-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 32 | Fmoc-Ser(tBu)-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 33 | Fmoc-Thr(tBu)-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 34 | Fmoc-Phe-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 35 | Fmoc-Thr(tBu)-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 36 | Fmoc-Gly-OH(6.00eq) | HATU(5.70eq)and DIEA(12.0eq) |
| 37 | Fmoc-Glu(tBu)-OH(6.00eq) | HOBT(6.00eq)and DIC(6.00eq) |
| 38 | Fmoc-Aib-OH(3.00eq) | HATU(2.85eq)and DIEA(6.00eq) |
| 39 | Boc-Tyr(tBu)-OH(3.00eq) | HATU(2.85eq)and DIEA(6.00eq) |
实施例1
1.长链脂肪酸的偶联：参考B-1合成步骤3.2完成以下片段的偶联:
| 序号 | 原料 | 偶联试剂 |
| 1 | Fmoc-AEEA-OH(3.00eq) | HATU(2.85eq)and DIEA(6.00eq) |
| 2 | Fmoc-AEEA-OH(3.00eq) | HATU(2.85eq)and DIEA(6.00eq) |
| 3 | Fmoc-Glu-OtBu(3.00eq) | HATU(2.85eq)and DIEA(6.00eq) |
| 4 | 20-(tBu)-二十烷二酸(3.00eq) | HATU(2.85eq)and DIEA(6.00eq) |
2.切割及粗肽干燥
2.1.按如下体积配置切割液
将干燥后的肽树脂加入到配好的切割液中，在摇床震荡2.5h，过滤，滤液加入到10倍体积冰异丙醚中，离心，再用异丙醚洗涤3次。在真空干燥2h得到粗肽，纯化得到多肽化合物WX-001，多肽分子量经ESI-MS进行确认，计算值为4811.4，检测值为4812.0。
实施例2
1.长链脂肪酸的偶联：参考B-1合成步骤3.2完成以下片段的偶联:
| 序号 | 原料 | 偶联试剂 |
| 1 | Fmoc-AEEA-OH(3.00eq) | HATU(2.85eq)and DIEA(6.00eq) |
| 2 | Fmoc-AEEA-OH(3.00eq) | HATU(2.85eq)and DIEA(6.00eq) |
| 3 | Fmoc-Glu-OtBu(3.00eq) | HATU(2.85eq)and DIEA(6.00eq) |
| 4 | Fmoc-Glu-OtBu(3.00eq) | HATU(2.85eq)and DIEA(6.00eq) |
| 5 | 20-(tBu)-二十烷二酸(3.00eq) | HATU(2.85eq)and DIEA(6.00eq) |
2.切割及粗肽干燥
2.1.按如下体积配置切割液
将干燥后的肽树脂加入到配好的切割液中，在摇床震荡2.5h，过滤，滤液加入到10倍体积冰异丙醚中，离心，再用异丙醚洗涤3次。在真空干燥2h得到粗肽，纯化得到多肽化合物WX-002。多肽分子量经ESI-MS进行确认，计算值为4940.5，检测值为4940.6。
实施例3
1.长链脂肪酸的偶联：参考B-1合成步骤3.2完成以下片段的偶联:
| 序号 | 原料 | 偶联试剂 |
| 1 | Fmoc-AEEA-OH(3.00eq) | HATU(2.85eq)and DIEA(6.00eq) |
| 2 | Fmoc-AEEA-OH(3.00eq) | HATU(2.85eq)and DIEA(6.00eq) |
| 3 | Fmoc-Glu-OtBu(3.00eq) | HATU(2.85eq)and DIEA(6.00eq) |
| 4 | Fmoc-Glu-OtBu(3.00eq) | HATU(2.85eq)and DIEA(6.00eq) |
| 5 | 20-(tBu)-十八烷二酸(3.00eq) | HATU(2.85eq)and DIEA(6.00eq) |
2.切割及粗肽干燥
2.1.按如下体积配置切割液
将干燥后的肽树脂加入到配好的切割液中，在摇床震荡2.5h，过滤，滤液加入到10倍体积冰异丙醚中，离心，再用异丙醚洗涤3次。在真空干燥2h得到粗肽，纯化得到多肽化合物WX-003。多肽分子量经ESI-MS进行确认，计算值为4912.5，检测值为4913.1。
实施例4
参考WX-001的合成，通过中间体B-2得到WX-004。多肽分子量经ESI-MS进行确认，计算值为4797.4，检测值为4797.3。
实施例5
参考WX-001的合成，通过中间体B-3得到WX-005。多肽分子量经ESI-MS进行确认，计算值为4869.5，检测值为4869.3。
实施例6
参考WX-001的合成，通过中间体B-4得到WX-006。多肽分子量经ESI-MS进行确认，计算值为4825.5，检测值为4825.5。
实施例7
参考WX-001的合成，通过中间体B-5得到WX-007。多肽分子量经ESI-MS进行确认，计算值为4811.4，检测值为4811.1。
实施例8
参考WX-001的合成，通过中间体B-6得到WX-008。多肽分子量经ESI-MS进行确认，计算值为4811.4，检测值为4811.4。
实施例9
参考WX-001的合成，通过中间体B-7得到WX-009。多肽分子量经ESI-MS进行确认，计算值为4854.5，检测值为4854.9。
实施例10
参考WX-001的合成，通过中间体B-8得到WX-010。多肽分子量经ESI-MS进行确认，计算值为4839.4，检测值为4839.6。
生物测试数据
实验例1：体外GLP-1R/GIPR激动活性测试
A：主要材料：
1)细胞株
该细胞株由上海药明康德构建。详情见下表。
| 靶点 | 宿主细胞 | 克隆 |
| GLP-1R | HEK293 | N/A |
| GIPR | CHO | N/A |
2)试剂与耗材
| 名称 | 批次. | 货号 | 厂家 |
| cAMP检测盒 | 29F | 62AM4PEJ | Cisbio |
| 1M HEPES | 2120919 | 15630-106 | Invitrogen |
| Hanks平衡盐溶液(HBSS) | 2185775 | 14025 | Invitrogen |
| 人血清白蛋白(HSA) | SLCF7301 | A1653-10G | 西格玛 |
| 酪蛋白 | SLCC9458 | C4765-10mL | 西格玛 |
| 3-异丁基-1-甲基黄嘌呤(IBMX) | STBF6061V | I5879-5G | 西格玛 |
| ECHO qualified 384孔板 | 0006433672 | PP-0200 | Labcyte |
| OptiPlate-384 | 8210-19481 | 6007299 | 珀金埃尔默 |
3)仪器
| 名称 | 型号 | 厂家 |
| EnVision | envision2014 | 珀金埃尔默 |
| Vi-cell counter | Vi-CELL TMXR Cell Viability Analyzer | 贝克曼 |
| Bravo | Bravo V11 | 安捷伦 |
| ECHO | ECHO 555 | Labcyte |
| Centrifuge | Allegra TM25R Centrifuge | 贝克曼 |
B.方法
1)实验材料
实验缓冲液
检测试剂制备
| 试剂 | 储存浓度 | 体积 | 终浓度 |
| 细胞裂解液 | 1x | 9.5mL | ≈1x |
| D2-cAMP溶液 | 40x | 250μL | 1x |
| cAMP-抗体溶液 | 40x | 250μL | 1x |
2)实验方法
a)制备化合物板：
待测化合物做10个点4倍稀释，起始浓度为30μM，Bravo完成稀释。
b)转移化合物：
1)使用Echo转移100nL化合物至OptiPlate-384plate。
2)将OptiPlate-384plate在1000rpm离心5秒。
c)细胞悬液的制备
1)将一支GLP-1R/GIPR细胞冻存管迅速置于37℃温水中解冻。
2)将细胞悬液转移至Transfer15mL离心管中，用10ml HBSS轻柔冲洗。
3)将离心管在1000rpm室温离心1分钟。
4)弃去上清。
5)轻柔打散底部细胞并再用10mL HBSS轻柔冲洗，离心沉降细胞，最后用实验缓冲液重悬细胞。
6)利用Vi-cell测量细胞密度与活度。
7)用实验缓冲液将GLP-1R/GIPR细胞浓度稀释至2.0*10
5/mL。
8)在OptiPlate-384plate中转入100nL稀释好的细胞悬液。
9)室温孵育30分钟。
d)加入检测试剂：
1)在OptiPlate-384plate空孔中加入10μL 800nM梯度稀释好的cAMP标准品。
2)加入10μL cAMP检测试剂。
3)用TopSeal-A film覆盖OptiPlate-384plate，室温孵育60分钟。
揭去TopSeal-A,在EnVision读数。
C实验结果
实验结果如表1所示。
表1体外GLP-1R/GIPR激动活性测试结果
结论：本发明化合物对GLP-1R/GIPR具有很强的激动活性。
实验例2：化合物大鼠药代动力学评价
A.实验目的
测试化合物在SD大鼠体内药代动力学
B.实验操作
以标准方案测试化合物皮下注射后的啮齿类动物药代特征，实验中候选化合物配成澄清溶液，给予大鼠单次皮下注射(SC,0.048mpk)。注射溶媒为柠檬酸盐缓冲液(20mM,pH＝7)。收集全血，制备得到血浆，以LC-MS/MS方法分析药物浓度，并用Phoenix WinNonlin软件计算药代参数。
C.实验结果
实验结果如表2所示。
表2药代动力学测试结果
| 化合物编号 | 最大血浆浓度(nM) | 半衰期T 1/2(h) | SC浓度积分AUC 0-72h(nM.hr) |
| WX001 | 22 | 12 | 728 |
| WX002 | 14 | 17 | 1316 |
| WX005 | 14 | 17 | 1329 |
| WX006 | 16 | 10 | 846 |
结论：本发明化合物具有优异的大鼠药代动力学性质。
实验例3：化合物小鼠药代动力学评价
A.实验目的
测试化合物在C57BL/6小鼠体内药代动力学
B.实验操作
以标准方案测试化合物静脉注射及皮下注射给药后的啮齿类动物药代特征，实验中候选化合物配成澄清溶液，给予小鼠单次静脉注射(IV,0.048mpk)及皮下注射给药(SC,0.048mpk)。静注溶媒为PBS缓冲液(pH＝7)，皮下注射溶媒为柠檬酸盐缓冲液(20mM,pH＝7)。收集全血，制备得到血浆，以LC-MS/MS方法分析药物浓度，并用Phoenix WinNonlin软件计算药代参数。
C.实验结果
实验结果如表3所示。
表3药代动力学测试结果
结论：本发明化合物具有优异的小鼠药代动力学性质。
实验例4：化合物食蟹猴药代动力学评价
A.实验目的
测试化合物在食蟹猴体内药代动力学
B.实验操作
以标准方案测试化合物静脉注射及皮下注射给药后的哺乳类动物药代特征，实验中候选化合物配成澄清溶液，给予食蟹猴单次皮下注射给药(SC,0.02mpk)。皮下注射溶媒为柠檬酸盐缓冲液(20mM,pH＝7)。收集全血，制备得到血浆，以LC-MS/MS方法分析药物浓度，并用Phoenix WinNonlin软件计算药代参数。
C.实验结果
实验结果如表4所示。
表4药代动力学测试结果
| 化合物编号 | C max(nM) | T max(h) | 半衰期T 1/2(h) | SC浓度积分AUC 0-240h(nM.hr) |
| WX001 | 30 | 21 | 80 | 3973 |
| WX002 | 33 | 29 | 81 | 4582 |
| WX005 | 31 | 40 | 98 | 4553 |
| WX006 | 30 | 29 | 72.4 | 3640 |
结论：本发明化合物具有优异的猴药代动力学性质。
实验例5：血浆稳定性测试(PLS)
A.实验目的
研究受试化合物在正常小鼠血浆中的稳定性。
B.实验操作
1.实验前，将凝结的冷冻血浆置于37℃的水浴锅中解冻。血浆4000rpm离心5min，如有血块，清除血块，将pH值调到7.4±0.1。
2.测试化合物溶液的制备：用DMSO稀释制得100μM的溶液。
3.98μL的空白对照血浆加入2μL的测试化合物溶液(100μM)，使得两者混合溶液的最终浓度达到2μM,将其置于37℃水浴条件下培养。
4.在每个时间点(0、10、30、60和120min)分别加入100μL H
3PO
4溶液和800μL终止溶液(200ng/mL甲糖宁和200ng/mL拉贝洛尔100％的甲醇溶液)来沉淀蛋白并充分混合。
5.样品在转速4000rpm下离心20min，从每孔取100μL上清液进行LC-MS/MS分析。
C.实验结果
实验结果如表5所示。
表5 PLS测试结果
| 化合物编号 | WX001 | WX002 | WX005 | WX006 |
| PLS(H/M)T 1/2(min) | >289/>289 | >289/>289 | >289/>289 | >289/>289 |
结论：本发明化合物具有优异的血浆稳定性。
实验例6：血浆蛋白结合度测试(PPB)
A.实验目的
研究受试化合物与人/小鼠血浆白蛋白的结合度。
B.实验操作
1.基质准备：实验当天，将血浆在冷水中解冻，并以3220rpm的速度离心5min，以去除所有血块。测量得到的血浆的pH值，并根据需要使用1％的磷酸或1N的氢氧化钠将其pH调整到7.4±0.1。
2.测试化合物的稀释步骤：测试化合物溶解在二甲基亚砜(DMSO)中，以制备浓度分别为10mM和2mM的原液。用98μL DMSO稀释2μL原液(2mM)，制得40μM的工作溶液。用240μL DMSO稀 释10μL原液，制得400μM对照化合物的工作溶液。将化合物的工作溶液(5μL)与空白基质(995μL)按1:200的比例混合均匀以制备负载基质。
3.分析步骤
3.1将等量的30μL负载基质(n＝2)转移至样品采集板，制备待测时间0(T0)样品用于残留测定。样品立即与相应的空白缓冲液进行匹配，最终体积为60μL，每孔中血浆与缓冲液的体积比为1:1。然后，测试化合物的T0样品分别加入60μL的4％H
3PO
4的H
2O和480μL含有内标物的终止液。然后将它们与其他样品一起储存在2-8℃下等待进一步处理。
3.2将剩余的血浆样品放在37±1℃的二氧化碳培养箱预培养30min。准备无蛋白样品(F样品)，负载基质的样品(230μL)都被转移到聚碳酸酯管(n＝2)中，并在37℃和155000×g(35000rpm)条件下超速离心4h。
3.3为了制备T样品(测试样品)，额外一份含基质样品转移到单独的96孔板(样品培育板)上，并在37℃下培育4h。
3.4离心结束后，从上清液第二层(上层以下)取30μL的无蛋白样品和30μL的T样品转移到新的样品收集板中。每个样品与相应的空白缓冲液或基质混合，最终体积为60μL，基质:缓冲液体积比1:1。在所有样品加入60μL 4％的H
3PO
4水溶液和480μL的终止溶液(含内标)。混合物在转速4000rpm下离心20min，取各样品上清液100μL进行LC-MS/MS分析。
C.实验结果
实验结果如表6所示。注：NA表示血浆蛋白结合度过高，在正常血浆蛋白浓度下未检出游离药物。
表6 PPB测试结果
| 化合物编号 | WX001 | WX002 | WX005 | WX006 |
| PPB％unbound(H/M) | NA/NA | NA/0.21％ | NA/0.22％ | NA/0.35％ |
结论：本发明化合物具有极高的血浆蛋白结合度。
实验例7：肾匀浆稳定性测试(BHS)
A.实验目的
研究受试化合物在正常小鼠肾匀浆中的稳定性。
B.实验操作
1.实验前，将冷冻肾匀浆放入37℃水浴中解冻。
2.测试化合物：用DMSO稀释10mM的原液，制备1mM的中间溶液；
3.测试化合物：用DMSO稀释1mM的中间溶液，制备50μM的加药溶液；
4.在每个时间点(0、10、30、60、120、180、240min)分别加入100μL的4％的H
3PO
4和800μL的终止液来沉淀蛋白并充分混合。
5.样品4000rpm离心10min，每孔取100μL上清液至板上。样品在800rpm下振荡约10min，然后提交LC-MS/MS测试。
C.实验结果
实验结果如表7所示。
表7 BHS测试结果
| 化合物编号 | WX001 | WX002 | WX005 | WX006 |
| BHS(T 1/2(min)) | 14 | 20 | 21 | 234 |
实验例8：化合物小鼠腹腔糖耐量(ipGTT)实验-体内药效评价
A.实验目的
研究受试化合物对于正常小鼠糖耐量的改善作用
B.实验操作
1.根据小鼠体重和血糖分组后，对每组动物分别注射待测化合物(0.3nmol/kg)和溶媒(20mM柠檬酸盐缓冲液)，过夜禁食，18小时后腹腔注射葡萄糖溶液(2g/kg,10mL/kg)；
2.使用血糖仪测定给葡萄糖后-60，0，15，30，60和120分钟的血糖浓度。
C.实验结果
实验结果如表8所示。
表8 ipGTT测试结果
| 化合物编号 | 溶媒 | WX001 | WX002 | WX005 | WX006 |
| 血糖AUC 0-120min(mmol.min) | 1916 | 1277 | 1150 | 1143 | 1164 |
结论：本发明化合物具有优异的糖耐量改善作用。
实验例9：在db/db小鼠中的药效研究-体内药效评价
A.实验目的
研究受试化合物对于II型糖尿病db/db小鼠的血糖控制作用
B.实验操作
1.db/db小鼠动物到达设施后，将其饲养于严格控制环境条件的动物饲养间中，饲养间的温度维持在20～24℃，湿度维持在30～70％。通过温湿度计对饲养间的温度和湿度进行实时监控，并且每天对温度和湿度 记录两次(上午和下午各1次)。动物饲养间的采光由一个电子定时开灯系统来控制，每天开灯12小时关灯12小时(上午7:00开，下午19:00关)。实验过程中，动物单笼饲养，并给每个笼中提供玩具。实验过程中动物自由采食(大小鼠生长/繁殖饲料)和饮水。
2.对每组动物分别皮下注射溶媒和待测化合物(15nmol/kg)，给药时间：早上9:30-11:00，给药频率一天一次，连续给药4周。
C.实验结果
实验结果如表9所示。
表9测试化合物在db/db小鼠中的降糖效果
| 化合物编号 | 溶媒 | WX001 | WX002 | WX005 | WX006 |
| ΔHbA1c％(连续给药四周后与第1天相比) | 15％ | -28％ | -21％ | -21％ | -23％ |
结论：本发明化合物在db/db小鼠中展现了优异的降糖药效。
实验例10：在DIO小鼠中的药效研究-体内药效评价
A.实验目的
研究受试化合物在DIO小鼠中的减重作用
B.实验操作
1.DIO小鼠动物到达药明康德设施后，将其饲养于严格控制环境条件的动物饲养间中，饲养间的温度维持在20～24℃，湿度维持在30～70％。通过温湿度计对饲养间的温度和湿度进行实时监控，并且每天对温度和湿度记录两次(上午和下午各1次)。动物饲养间的采光由一个电子定时开灯系统来控制，每天开灯12小时关灯12小时(上午7:00开，下午19:00关)。实验过程中，动物单笼饲养，并给每个笼中提供玩具。实验过程中动物自由采食(大小鼠生长/繁殖饲料)和饮水。
2.对每组动物分别皮下注射溶媒和待测化合物(10nmol/kg)，给药时间：早上9:30，给药频率三天一次，给药周期为22天。
C.实验结果
实验结果如表10所示。
表10测试化合物在DIO小鼠中的药效
| 化合物编号 | 溶媒 | WX001 | WX002 | WX005 | WX006 |
| Δ体重％(22天后与第1天相比) | 0.2％ | -29％ | -26％ | -24％ | -32％ |
| Δ脂肪组织％(22天后与第1天相比) | 2％ | -54％ | -48％ | -47％ | -62％ |
| Δ非脂肪组织％(22天后与第1天相比) | -3％ | -11％ | -10％ | -8％ | -18％ |
结论：本发明化合物在DIO小鼠中展现了优异的减重药效。
Claims (11)
- 下式化合物或其药学上可接受的盐，SEQ ID NO1：YAibEGT FTSDY SIAibLD KK 1AZ 0K 0Z 1FZ 2E 0W LZ 3AGG PSSGA PPPS 0SEQ ID NO2：YAibEGT FTSDY SIAibLD KE 0AQK 0AFVK 1W LIAGG PSSGA PPPS 0SEQ ID NO3：YAibEGT FTSDY SIE 0LD KK 0AQK 1AFVQW LIAGG PSSGA PPPS 0SEQ ID NO4：YAibEGT FTSDY SIE 0LD K 0IAQK 1AFVQW LIAGG PSSGA PPPS 0其中，Aib的结构为
S 0选自
Z 0选自谷氨酰胺(Q)和天冬酰胺(N)；Z 1选自丙氨酸(A)和谷氨酸(E)；Z 2选自缬氨酸(V)和异亮氨酸(I)；Z 3选自异亮氨酸(I)和亮氨酸(L)；E 0和K 0表示谷氨酸侧链上的羧基与赖氨酸侧链上的氨基共同构成内酰胺，此时K 0Z 1FZ 2E 0的结构为E 0AQK 0的结构为
E 0LDKK 0的结构为
E 0LDK 0的结构为
K 1表示赖氨酸侧链上的氨基与-X-X 1-X 2相连，其结构为
X选自
R 1和R 2分别独立地选自H和CH 3；X 1选自
X 2选自
m、n和p分别独立地选自2和3；s选自2和3；q选自15、16、17、18和19。 - 根据权利要求1所述化合物或其药学上可接受的盐，其选自：
其中，Aib、Z 0、Z 1、Z 2、Z 3和K 1如权利要求1所定义。 - 根据权利要求1或2所述化合物或其药学上可接受的盐，其中，X选自-C(＝O)-CH 2-。
- 根据权利要求1或2所述化合物或其药学上可接受的盐，其中，m、n和p分别独立地选自2。
- 根据权利要求1或2所述化合物或其药学上可接受的盐，其中，s独立地选自2。
- 根据权利要求1或2所述化合物或其药学上可接受的盐，其中，q独立地选自15和17。
- 根据权利要求1或2所述化合物或其药学上可接受的盐，其中，X 1选自
- 根据权利要求1或2所述化合物或其药学上可接受的盐，其中，X 2选自
- 根据权利要求1或2所述化合物或其药学上可接受的盐，其中，-X-X 1-X 2选自
- 下式所示化合物或其药学上可接受的盐，
- 根据权利要求1～10任意一项所述的化合物或其药学上可接受的盐在制备治疗肥胖症及糖尿病的药物上的应用。
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US11744873B2 (en) | 2021-01-20 | 2023-09-05 | Viking Therapeutics, Inc. | Compositions and methods for the treatment of metabolic and liver disorders |
| WO2023193727A1 (zh) * | 2022-04-07 | 2023-10-12 | 广东众生睿创生物科技有限公司 | 多肽在制备治疗和/或预防糖尿病及肥胖症及其相关疾病药物中的制药用途 |
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- 2021-12-02 AU AU2021391241A patent/AU2021391241B2/en active Active
- 2021-12-02 EP EP21900082.5A patent/EP4257597A1/en active Pending
- 2021-12-02 US US18/039,582 patent/US20240067702A1/en active Pending
- 2021-12-02 WO PCT/CN2021/135180 patent/WO2022117056A1/zh active Application Filing
- 2021-12-02 CA CA3200881A patent/CA3200881A1/en active Pending
- 2021-12-02 BR BR112023010891A patent/BR112023010891A2/pt unknown
- 2021-12-02 CN CN202180080890.XA patent/CN116615222A/zh active Pending
- 2021-12-02 MX MX2023006419A patent/MX2023006419A/es unknown
- 2021-12-02 JP JP2023533785A patent/JP2023552362A/ja active Pending
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| CN108699125A (zh) * | 2015-12-31 | 2018-10-23 | 韩美药品株式会社 | 激活胰高血糖素、glp-1和gip受体的三重激活剂 |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US11744873B2 (en) | 2021-01-20 | 2023-09-05 | Viking Therapeutics, Inc. | Compositions and methods for the treatment of metabolic and liver disorders |
| WO2023193727A1 (zh) * | 2022-04-07 | 2023-10-12 | 广东众生睿创生物科技有限公司 | 多肽在制备治疗和/或预防糖尿病及肥胖症及其相关疾病药物中的制药用途 |
Also Published As
| Publication number | Publication date |
|---|---|
| EP4257597A1 (en) | 2023-10-11 |
| CA3200881A1 (en) | 2022-06-09 |
| MX2023006419A (es) | 2023-08-15 |
| BR112023010891A2 (pt) | 2023-10-17 |
| AU2021391241A1 (en) | 2023-07-20 |
| AU2021391241B2 (en) | 2024-05-09 |
| JP2023552362A (ja) | 2023-12-15 |
| CN116615222A (zh) | 2023-08-18 |
| CO2023008676A2 (es) | 2023-09-29 |
| KR20230116894A (ko) | 2023-08-04 |
| US20240067702A1 (en) | 2024-02-29 |
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