CN114369139A - Preparation method of snake venom-like tripeptide - Google Patents
Preparation method of snake venom-like tripeptide Download PDFInfo
- Publication number
- CN114369139A CN114369139A CN202011097855.2A CN202011097855A CN114369139A CN 114369139 A CN114369139 A CN 114369139A CN 202011097855 A CN202011097855 A CN 202011097855A CN 114369139 A CN114369139 A CN 114369139A
- Authority
- CN
- China
- Prior art keywords
- boc
- dab
- ala
- beta
- pro
- 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.)
- Granted
Links
- 238000002360 preparation method Methods 0.000 title claims abstract description 13
- 241000270295 Serpentes Species 0.000 title claims description 21
- LIWKOFAHRLBNMG-FQEVSTJZSA-N (2s)-2-(9h-fluoren-9-ylmethoxycarbonylamino)-4-[(2-methylpropan-2-yl)oxycarbonylamino]butanoic acid Chemical compound C1=CC=C2C(COC(=O)N[C@@H](CCNC(=O)OC(C)(C)C)C(O)=O)C3=CC=CC=C3C2=C1 LIWKOFAHRLBNMG-FQEVSTJZSA-N 0.000 claims abstract description 36
- 238000000034 method Methods 0.000 claims abstract description 32
- WGQKYBSKWIADBV-UHFFFAOYSA-N benzylamine Chemical compound NCC1=CC=CC=C1 WGQKYBSKWIADBV-UHFFFAOYSA-N 0.000 claims abstract description 25
- 239000003998 snake venom Substances 0.000 claims abstract description 25
- TVWATMRQKCTKAU-UHFFFAOYSA-N (2,5-dioxopyrrolidin-1-yl) 3-[(2-methylpropan-2-yl)oxycarbonylamino]propanoate Chemical compound CC(C)(C)OC(=O)NCCC(=O)ON1C(=O)CCC1=O TVWATMRQKCTKAU-UHFFFAOYSA-N 0.000 claims abstract description 16
- ONIBWKKTOPOVIA-BYPYZUCNSA-N L-Proline Chemical compound OC(=O)[C@@H]1CCCN1 ONIBWKKTOPOVIA-BYPYZUCNSA-N 0.000 claims abstract description 12
- 238000000926 separation method Methods 0.000 claims abstract description 7
- 125000003088 (fluoren-9-ylmethoxy)carbonyl group Chemical group 0.000 claims abstract description 6
- -1 Boc groups Chemical group 0.000 claims abstract description 6
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 97
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 54
- 238000006243 chemical reaction Methods 0.000 claims description 54
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 claims description 36
- 235000019439 ethyl acetate Nutrition 0.000 claims description 33
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 claims description 26
- 239000007864 aqueous solution Substances 0.000 claims description 23
- 239000002904 solvent Substances 0.000 claims description 23
- FDKXTQMXEQVLRF-ZHACJKMWSA-N (E)-dacarbazine Chemical compound CN(C)\N=N\c1[nH]cnc1C(N)=O FDKXTQMXEQVLRF-ZHACJKMWSA-N 0.000 claims description 21
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims description 21
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 claims description 21
- 239000000243 solution Substances 0.000 claims description 20
- 239000007787 solid Substances 0.000 claims description 16
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 16
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 15
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 15
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 14
- 229910000029 sodium carbonate Inorganic materials 0.000 claims description 13
- 239000003795 chemical substances by application Substances 0.000 claims description 12
- YEDUAINPPJYDJZ-UHFFFAOYSA-N 2-hydroxybenzothiazole Chemical compound C1=CC=C2SC(O)=NC2=C1 YEDUAINPPJYDJZ-UHFFFAOYSA-N 0.000 claims description 11
- NPZTUJOABDZTLV-UHFFFAOYSA-N hydroxybenzotriazole Substances O=C1C=CC=C2NNN=C12 NPZTUJOABDZTLV-UHFFFAOYSA-N 0.000 claims description 11
- 239000003960 organic solvent Substances 0.000 claims description 11
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical group [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 claims description 10
- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 claims description 10
- LMDZBCPBFSXMTL-UHFFFAOYSA-N 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide Substances CCN=C=NCCCN(C)C LMDZBCPBFSXMTL-UHFFFAOYSA-N 0.000 claims description 9
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 claims description 9
- 150000001413 amino acids Chemical class 0.000 claims description 9
- FPQQSJJWHUJYPU-UHFFFAOYSA-N 3-(dimethylamino)propyliminomethylidene-ethylazanium;chloride Chemical compound Cl.CCN=C=NCCCN(C)C FPQQSJJWHUJYPU-UHFFFAOYSA-N 0.000 claims description 8
- QOSSAOTZNIDXMA-UHFFFAOYSA-N Dicylcohexylcarbodiimide Chemical compound C1CCCCC1N=C=NC1CCCCC1 QOSSAOTZNIDXMA-UHFFFAOYSA-N 0.000 claims description 8
- PFKFTWBEEFSNDU-UHFFFAOYSA-N carbonyldiimidazole Chemical compound C1=CN=CN1C(=O)N1C=CN=C1 PFKFTWBEEFSNDU-UHFFFAOYSA-N 0.000 claims description 8
- 230000018044 dehydration Effects 0.000 claims description 8
- 238000006297 dehydration reaction Methods 0.000 claims description 8
- 238000006482 condensation reaction Methods 0.000 claims description 7
- 239000003208 petroleum Substances 0.000 claims description 7
- 238000005406 washing Methods 0.000 claims description 7
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 6
- 230000002378 acidificating effect Effects 0.000 claims description 6
- 239000012046 mixed solvent Substances 0.000 claims description 5
- 229910000030 sodium bicarbonate Inorganic materials 0.000 claims description 5
- 239000002585 base Substances 0.000 claims description 4
- MVEAAGBEUOMFRX-UHFFFAOYSA-N ethyl acetate;hydrochloride Chemical compound Cl.CCOC(C)=O MVEAAGBEUOMFRX-UHFFFAOYSA-N 0.000 claims description 4
- 239000007788 liquid Substances 0.000 claims description 4
- 239000007821 HATU Substances 0.000 claims description 3
- 238000002156 mixing Methods 0.000 claims description 3
- GSNUFIFRDBKVIE-UHFFFAOYSA-N DMF Natural products CC1=CC=C(C)O1 GSNUFIFRDBKVIE-UHFFFAOYSA-N 0.000 claims description 2
- 239000003513 alkali Substances 0.000 claims description 2
- 238000009833 condensation Methods 0.000 claims description 2
- 230000005494 condensation Effects 0.000 claims description 2
- 239000011736 potassium bicarbonate Substances 0.000 claims description 2
- 229910000028 potassium bicarbonate Inorganic materials 0.000 claims description 2
- BWHMMNNQKKPAPP-UHFFFAOYSA-L potassium carbonate Substances [K+].[K+].[O-]C([O-])=O BWHMMNNQKKPAPP-UHFFFAOYSA-L 0.000 claims description 2
- 229910000027 potassium carbonate Inorganic materials 0.000 claims description 2
- TYJJADVDDVDEDZ-UHFFFAOYSA-M potassium hydrogencarbonate Chemical compound [K+].OC([O-])=O TYJJADVDDVDEDZ-UHFFFAOYSA-M 0.000 claims description 2
- 239000011541 reaction mixture Substances 0.000 claims 2
- 239000003929 acidic solution Substances 0.000 claims 1
- 239000012670 alkaline solution Substances 0.000 claims 1
- 238000000746 purification Methods 0.000 abstract description 10
- 239000003153 chemical reaction reagent Substances 0.000 abstract description 6
- 238000004519 manufacturing process Methods 0.000 abstract description 6
- 238000004128 high performance liquid chromatography Methods 0.000 abstract description 4
- 238000003786 synthesis reaction Methods 0.000 description 20
- 230000015572 biosynthetic process Effects 0.000 description 19
- 238000003756 stirring Methods 0.000 description 16
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 13
- 238000004809 thin layer chromatography Methods 0.000 description 12
- NQRYJNQNLNOLGT-UHFFFAOYSA-N Piperidine Chemical compound C1CCNCC1 NQRYJNQNLNOLGT-UHFFFAOYSA-N 0.000 description 10
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 description 9
- 238000001035 drying Methods 0.000 description 8
- 238000001914 filtration Methods 0.000 description 8
- 238000012544 monitoring process Methods 0.000 description 6
- 150000003839 salts Chemical class 0.000 description 6
- 238000005160 1H NMR spectroscopy Methods 0.000 description 5
- 230000002194 synthesizing effect Effects 0.000 description 5
- IAZDPXIOMUYVGZ-WFGJKAKNSA-N Dimethyl sulfoxide Chemical compound [2H]C([2H])([2H])S(=O)C([2H])([2H])[2H] IAZDPXIOMUYVGZ-WFGJKAKNSA-N 0.000 description 4
- 238000010438 heat treatment Methods 0.000 description 4
- 108090000765 processed proteins & peptides Proteins 0.000 description 4
- 239000007832 Na2SO4 Substances 0.000 description 3
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical class [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 3
- 150000001875 compounds Chemical class 0.000 description 3
- 238000007796 conventional method Methods 0.000 description 3
- 238000001816 cooling Methods 0.000 description 3
- 238000002425 crystallisation Methods 0.000 description 3
- 230000008025 crystallization Effects 0.000 description 3
- 238000004090 dissolution Methods 0.000 description 3
- 239000012535 impurity Substances 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 239000012071 phase Substances 0.000 description 3
- 229910052938 sodium sulfate Inorganic materials 0.000 description 3
- WMFOQBRAJBCJND-UHFFFAOYSA-M Lithium hydroxide Chemical compound [Li+].[OH-] WMFOQBRAJBCJND-UHFFFAOYSA-M 0.000 description 2
- DTQVDTLACAAQTR-UHFFFAOYSA-N Trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 description 2
- 239000008346 aqueous phase Substances 0.000 description 2
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 2
- 239000002537 cosmetic Substances 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 239000007791 liquid phase Substances 0.000 description 2
- 125000000956 methoxy group Chemical group [H]C([H])([H])O* 0.000 description 2
- 125000006239 protecting group Chemical group 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- BDNKZNFMNDZQMI-UHFFFAOYSA-N 1,3-diisopropylcarbodiimide Chemical compound CC(C)N=C=NC(C)C BDNKZNFMNDZQMI-UHFFFAOYSA-N 0.000 description 1
- ASOKPJOREAFHNY-UHFFFAOYSA-N 1-Hydroxybenzotriazole Chemical compound C1=CC=C2N(O)N=NC2=C1 ASOKPJOREAFHNY-UHFFFAOYSA-N 0.000 description 1
- GETXTESSLWBSTI-UHFFFAOYSA-N 1-[3-(2H-triazol-4-yl)propyl]cyclohexa-2,4-dien-1-ol Chemical compound OC1(CC=CC=C1)CCCC=1N=NNC=1 GETXTESSLWBSTI-UHFFFAOYSA-N 0.000 description 1
- WCFJUSRQHZPVKY-UHFFFAOYSA-N 3-[(2-methylpropan-2-yl)oxycarbonylamino]propanoic acid Chemical compound CC(C)(C)OC(=O)NCCC(O)=O WCFJUSRQHZPVKY-UHFFFAOYSA-N 0.000 description 1
- 101000761020 Dinoponera quadriceps Poneritoxin Proteins 0.000 description 1
- 102000019315 Nicotinic acetylcholine receptors Human genes 0.000 description 1
- 108050006807 Nicotinic acetylcholine receptors Proteins 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- 241000271897 Viperidae Species 0.000 description 1
- 101800002700 Waglerin-1 Proteins 0.000 description 1
- 150000001242 acetic acid derivatives Chemical class 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 150000007513 acids Chemical class 0.000 description 1
- 230000004075 alteration Effects 0.000 description 1
- 125000003277 amino group Chemical group 0.000 description 1
- 239000005557 antagonist Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000012964 benzotriazole Substances 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- BGRWYRAHAFMIBJ-UHFFFAOYSA-N diisopropylcarbodiimide Natural products CC(C)NC(=O)NC(C)C BGRWYRAHAFMIBJ-UHFFFAOYSA-N 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 238000009499 grossing Methods 0.000 description 1
- 150000003840 hydrochlorides Chemical class 0.000 description 1
- 238000009776 industrial production Methods 0.000 description 1
- XETISTBVDHNRBD-FJXQXJEOSA-N methyl (2s)-2-amino-4-[(2-methylpropan-2-yl)oxycarbonylamino]butanoate;hydrochloride Chemical compound Cl.COC(=O)[C@@H](N)CCNC(=O)OC(C)(C)C XETISTBVDHNRBD-FJXQXJEOSA-N 0.000 description 1
- HQEIPVHJHZTMDP-JEDNCBNOSA-N methyl (2s)-pyrrolidine-2-carboxylate;hydrochloride Chemical compound Cl.COC(=O)[C@@H]1CCCN1 HQEIPVHJHZTMDP-JEDNCBNOSA-N 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 210000003205 muscle Anatomy 0.000 description 1
- 239000012074 organic phase Substances 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 239000000741 silica gel Substances 0.000 description 1
- 229910002027 silica gel Inorganic materials 0.000 description 1
- 239000007790 solid phase Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 238000010189 synthetic method Methods 0.000 description 1
- 239000002435 venom Substances 0.000 description 1
- 231100000611 venom Toxicity 0.000 description 1
- 210000001048 venom Anatomy 0.000 description 1
- 239000002821 viper venom Substances 0.000 description 1
- 230000037303 wrinkles Effects 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
- C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
- C07K5/08—Tripeptides
- C07K5/0802—Tripeptides with the first amino acid being neutral
- C07K5/0804—Tripeptides with the first amino acid being neutral and aliphatic
- C07K5/0806—Tripeptides with the first amino acid being neutral and aliphatic the side chain containing 0 or 1 carbon atoms, i.e. Gly, Ala
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/55—Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups
Abstract
The invention discloses a preparation method of snake venom tripeptide, which comprises the steps of reacting Boc-beta-Ala-OSu with L-Pro-OH to obtain Boc-beta-Ala-Pro-OH, dehydrating and condensing Fmoc-dab (Boc) -OH with benzylamine, removing Fmoc groups to obtain H-dab (Boc) -NHBn, dehydrating and condensing Boc-beta-Ala-Pro-OH with H-dab (Boc) -NHBn to obtain Boc-beta-Ala-Pro-dab (Boc) -NHBn, and finally removing Boc groups from Boc-beta-Ala-Pro-dab (Boc) -NHBn to obtain snake venom tripeptide. The method has the advantages of high efficiency, convenience, high yield, simple post-treatment, no need of HPLC separation and purification, capability of obtaining a high-purity product, low reagent consumption, capability of greatly reducing the production cost and suitability for practical production and application.
Description
Technical Field
The invention relates to the field of chemical synthesis, in particular to a preparation method of snake venom-like tripeptide.
Background
The snake venom-like tripeptide (H-beta-Ala-Pro-DAB-NH-Bzl) is a small peptide that mimics the activity of the snake venom toxin WaglerinI, which is found in the venom of the Temple Viper venom (Tripidolaemus wagleri) and is an antagonist of the muscle nicotinic acetylcholine receptor (nmAChR). H-beta-Ala-Pro-DAB-NH-Bzl was originally synthesized by high-tech biochemical technology from DSM imperial Imperial group Quanzison company PENTAPHARM by simulating fragments of effective molecular protein (Waglerin1), has a chemical structure similar to that of viper serum, but is safer and more effective than botulinum, so that the H-beta-Ala-Pro-DAB-NH-Bzl can be safely applied to cosmetics and has excellent skin smoothing and quick wrinkle removing performances.
At present, most of conventional methods for synthesizing snake venom tripeptide (H-beta-Ala-Pro-DAB-NH-Bzl) adopt a solid-phase and liquid-phase combined method, the synthesis process is relatively complicated and high in cost, and in order to simplify the process to a certain extent, researchers develop a method for synthesizing the snake venom tripeptide in a full liquid phase, for example, patent CN107936108A discloses the following synthesis route:
reacting Boc-beta-Ala-OH, N-ethyl-5-phenylisoxazole-3' -sulfonic acid inner salt and H-Pro-OMe-HCl to obtain Boc-beta-Ala-Pro-OMe, and reacting the Boc-beta-Ala-Pro-OMe with LiOH to obtain Boc-beta-Ala-Pro-OH; then synthesizing Boc-beta-Ala-Pro DAB (Boc) -OH with Boc-beta-Ala-Pro-OH and H-DAB (Boc) -OMe-HCl by the same method; and reacting Boc-beta-Ala-Pro-DAB (Boc) -OH, 1-hydroxy phenylpropyl triazole, N-diisopropyl carbodiimide and benzylamine to obtain Boc-beta-Ala-Pro-DAB (Boc) -NH-Bzl, finally removing Boc protecting groups by trifluoroacetic acid, and separating and purifying by HPLC to obtain snake venom tripeptide with purity of about 98%, wherein the total yield is only about 38%, the synthetic route is long, the treatment steps are complicated, and the using amount of a plurality of reagents in the synthetic process is large.
Patent CN107857797A discloses another method for synthesizing snake venom tripeptide, which comprises the following steps:
the total yield of the method is only about 30 percent, and the snake venom-like tripeptide product with higher purity can be obtained after HPLC separation and purification treatment, and the method has long synthesis steps and complex operation.
Therefore, the problems of low yield, complex synthesis process and the like of the conventional method for synthesizing the snake venom tripeptide directly cause higher cost in actual production.
Therefore, the synthetic method of the snake-like tripeptide with simple development process, high yield and low cost has important significance for the production of the snake-like tripeptide and the application of the snake-like tripeptide in the field of cosmetics.
Disclosure of Invention
The invention mainly solves the technical problem of providing a preparation method of the snake venom-like tripeptide, which can efficiently synthesize the snake venom-like tripeptide and has simple process and high yield.
In order to solve the technical problems, the invention adopts a technical scheme that:
the invention provides a preparation method of Boc-beta-Ala-Pro-dab (Boc) -NHBn, which comprises the following steps: the amino acid dehydration condensation reaction of Boc-beta-Ala-Pro-OH and H-dab (Boc) -NHBn is carried out to obtain:
further, Boc-beta-Ala-Pro-OH: the molar use ratio of H-dab (Boc) -NHBn is 1.0-1.5: 1.0, preferably 1.1: 1.0.
further, the reaction conditions include a first condensing agent; further, the first condensing agent is selected from one or more of DIC, DCC and EDCI, preferably DIC;
further, the first condensing agent: the molar use ratio of H-dab (Boc) -NHBn is 1.0-1.5: 1.0, preferably 1.2: 1.0.
further, the solvent of the reaction is a first solvent, and the first solvent is selected from one or more of DCM, EtOAc and THF, and is preferably DCM.
Further, H-dab (Boc) -NHBn: the mass ratio of the first solvent is 1: 15-30; further 1: 20.
further, after the reaction is finished, the reaction solution is washed by alkaline aqueous solution and acidic aqueous solution and water respectively, solid-liquid separation is carried out after washing, and the solid is separated by ethyl acetate: the mass ratio of petroleum ether is 0.8-1.2: 0.8-1.2 of organic solvent crystallization purification.
The crystallization purification is a commonly used purification method in the synthesis process, namely, the solid is completely dissolved in the solvent by means of heating and the like, and then the solid is precipitated by means of cooling and the like.
The basic aqueous solution refers to an aqueous solution that is basic, and the acidic aqueous solution refers to an aqueous solution that is acidic, and any basic aqueous solution or acidic aqueous solution is suitable as long as it does not react with the product Boc-beta-Ala-Pro-dab (Boc) -NHBn, without limiting the solute.
The alkaline aqueous solution is a sodium carbonate aqueous solution with the mass fraction of 2-10%, and preferably a sodium carbonate aqueous solution with the mass fraction of 5%.
The acidic aqueous solution is 2-10% by mass of citric acid aqueous solution, and preferably 5% by mass of citric acid aqueous solution.
Further, the mass usage amount of the organic solvent is 15-25 times, and further 20 times of the mass of H-dab (Boc) -NHBn.
In the invention, the crystallization and purification are carried out by mixing the solid and the organic solvent, heating until the solid is completely dissolved, and then cooling to separate out the solid. In general, the solid can be completely dissolved by heating to about 60 ℃.
In a specific embodiment of the invention, the solid and ethyl acetate: the mass ratio of petroleum ether is 1:1, heating until the solid is completely dissolved, and then cooling to separate out the solid.
Further, the H-dab (Boc) -NHBn is obtained by performing amino acid dehydration condensation reaction on Fmoc-dab (Boc) -OH and benzylamine and then removing Fmoc group:
the amino acid molecules are bonded by the carboxyl group (-COOH) of one amino acid molecule and the amino group (-NH) of another amino acid molecule2) The connection is combined, and one molecule of water is removed at the same time, and the combination mode is called amino acid dehydration condensation.
Further, Fmoc-dab (boc) -OH: the molar use ratio of benzylamine is 0.8-1.2: 0.8 to 1.2, preferably 1: 1.
further, the conditions for the dehydration condensation reaction of the amino acid of Fmoc-dab (Boc) -OH with benzylamine include a second condensing agent; further, the second condensing agent is selected from one or more of DIC, DCC, EDCI, HOBT, CDI, HBTU and HATU, preferably DIC and/or HOBT, or EDCI; more preferably DIC and HOBT, DIC: the HOBT molar use ratio is 1: 1.
further, the second condensing agent: the molar ratio of Fmoc-dab (Boc) -OH is 1.0-3.0: 1.0, preferably 1.2 to 2.4: 1.0.
further, the solvent for the amino acid dehydration condensation reaction of Fmoc-dab (Boc) -OH and benzylamine is a second solvent, and the second solvent is selected from one or more of DCM, THF and acetonitrile, preferably DCM and/or THF, and more preferably DCM.
Further, Fmoc-dab (boc) -OH: the mass ratio of the second solvent is 1: 5-15, preferably 1: 10.
removal of the Fmoc group is a common removal of protecting groups and methods known in the art for removing Fmoc groups are applicable to the present invention.
In a specific embodiment of the invention, Fmoc-dab (Boc) -NHBn, a third solvent and piperidine are mixed for reaction, wherein the solvent is selected from one or more of methanol, DCM and THF, and methanol is preferred.
Further, Fmoc-dab (Boc) -NHBn: the mass usage ratio of the third solvent is 1.0: 5-20, preferably 1.0: 10.
further, Fmoc-dab (Boc) -NHBn: the molar ratio of the piperidine used is 1.0: 2-10, preferably 1.0: 5.
further, after removing the solvent from the reacted system, mixing the residue with dilute hydrochloric acid, extracting with DCM and/or ethyl acetate, adjusting the aqueous phase to be alkaline, extracting with ethyl acetate, and removing the solvent from the organic phase to obtain the product H-dab (Boc) -NHBn without Fmoc groups.
In a particular embodiment of the invention, the aqueous phase is adjusted to alkaline by adjusting it to a pH of 10 with sodium carbonate.
Further, the Boc-beta-Ala-Pro-OH is obtained by reacting Boc-beta-Ala-OSu with L-Pro-OH as follows:
further, Boc-beta-Ala-OSu: the molar use ratio of L-Pro-OH is 1.0: 1.0-1.5, preferably 1; 1.2;
further, the conditions for the reaction of Boc-beta-Ala-OSu with L-Pro-OH include a base; further, the base is selected from NaHCO3、KHCO3、Na2CO3、K2CO3Preferably NaHCO3；
Further, Boc-beta-Ala-OSu: the molar ratio of the alkali is 1.0: 1.5 to 3, preferably 1.0: 2.0.
further, the solvent for the reaction of Boc-beta-Ala-OSu with L-Pro-OH was water: the molar use ratio of the organic solvent is 0.8-1.2: 0.8-1.2, preferably water: the molar ratio of the organic solvent is 1:1, wherein the organic solvent is selected from one or more of tetrahydrofuran, acetonitrile, acetone, methanol, ethanol and DMF, preferably one or more of tetrahydrofuran, acetonitrile and acetone, and more preferably tetrahydrofuran;
further, Boc-beta-Ala-OSu: the mass ratio of the mixed solvent is 1: 5-15, preferably 1: 10.
the invention also provides a method for preparing snake venom tripeptide dihydrochloride, which is obtained by removing Boc group from Boc-beta-Ala-Pro-dab (Boc) -NHBn; wherein Boc-beta-Ala-Pro-dab (Boc) -NHBn is prepared by any of the methods described above.
Further, Boc-beta-Ala-Pro-dab (Boc) -NHBn is mixed with ethyl acetate, HCl ethyl acetate solution is added into the solution in batches, and solid-liquid separation is carried out after the reaction is finished to obtain solid matter which is snake venom tripeptide dihydrochloride.
The batch addition refers to non-one-time addition, and can be realized by adding the raw materials at a certain or different feeding speed within a period of time, or by dividing the raw materials to be added into a plurality of parts and adding the parts for a plurality of times within a certain period of time.
Further, Boc-beta-Ala-Pro-dab (Boc) -NHBn: the mass usage amount of the ethyl acetate is 1: 5-15, preferably 1: 10; Boc-beta-Ala-Pro-dab (Boc) -NHBn: the mass usage ratio of the HCl ethyl acetate solution is 1: 5-15, preferably 1: 10.
the invention also provides application of the snake venom-like tripeptide dihydrochloride in preparation of a snake venom-like tripeptide related product, wherein the snake venom-like tripeptide dihydrochloride is obtained by the preparation method.
The snake venom tripeptide related product refers to a product containing compounds in different forms of snake venom tripeptides, such as snake venom tripeptides, salts in different forms of snake venom tripeptides and the like, wherein the salts can be hydrochlorides, acetates and the like.
After the snake venom-like tripeptide dihydrochloride is prepared by the method, the snake venom-like tripeptide dihydrochloride can be processed by the conventional method to obtain various forms of compounds of the snake venom-like tripeptide, for example, different acids are adopted to be processed and converted into other salts (such as snake venom-like tripeptide diacetate), and the other salts are processed by alkalization to obtain the snake venom-like tripeptide which is used as a product; or further processing the snake venom tripeptide and/or salts thereof in different forms to obtain products, such as skin care products, medicines, etc. The snake venom-like tripeptide dihydrochloride prepared by the method belongs to the protection scope of the invention as long as the snake venom-like tripeptide dihydrochloride is prepared in the preparation process of the product.
The invention has the beneficial effects that:
(1) the method adopts a new synthetic route, can obtain the product with the purity of more than 98 percent without HPLC separation and purification, and has the advantages of simple process, convenient operation, small reagent dosage and high yield.
(2) The method has high efficiency and convenience in synthesis process, can greatly reduce the production cost, is more suitable for practical production and application, is suitable for large-scale industrial production, and is worthy of popularization.
The following abbreviations have the meanings indicated below:
DCC: dicyclohexylcarbodiimide;
DIC: diisopropylcarbodiimide;
EDCI: 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide;
HATU: 2- (7-benzotriazole oxide) -N, N' -tetramethyluronium hexafluorophosphate;
HBTU: o-benzotriazol-tetramethylurea hexafluorophosphate;
HOBT: 1-hydroxybenzotriazole;
CDI: n, N' -carbonyldiimidazole;
DCM: dichloromethane;
DMF: n, N-dimethylformamide;
THF: tetrahydrofuran.
Detailed Description
The technical solutions of the present invention are described clearly and completely below, and it is obvious that the described embodiments are some, not all embodiments of the present invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
The yields in the examples of the present invention are all molar yields.
In the examples, "eq" means equivalent, which means a multiple of the molar amount of the other reagents on the basis of a certain reagent (i.e., 1eq), "n-fold parts by weight" means an amount of 1eq by mass of the reagent, n-fold.
For example, "to a reaction vessel, Fmoc-dab (Boc) -OH (1eq) and dichloromethane (10-fold parts by weight) are added, followed by dissolution and subsequent addition of DIC (1.2 eq)" means that the molar amount of DIC is 1.2 times that of Fmoc-dab (Boc) -OH, and that if the molar amount of Fmoc-dab (Boc) -OH is 1mol, the molar amount of DIC is 1.2 mol; the mass dosage of the dichloromethane is 10 times that of the Fmoc-dab (Boc) -OH, and if the mass dosage of the Fmoc-dab (Boc) -OH is 1kg, the dosage of the dichloromethane is 10 kg; the same applies to the rest cases.
The synthesis scale of the scheme recorded in the embodiment of the invention is 10kg per batch, namely the mass of the snake venom-like tripeptide dihydrochloride prepared in each batch is about 10 kg.
The synthetic route is as follows:
step 1: Boc-beta-Ala-Pro-OH Synthesis:
step 2: synthesis of H-dab (Boc) -NHBn:
and step 3: synthesis of snake venom peptides:
the specific effects of the preparation process according to the invention will be illustrated by the following different examples:
example 1
(1) Synthesis of Boc-beta-Ala-Pro-OH
Adding L-Pro-OH (1.2eq), water (5 times of the weight of the mixture) and NaHCO into a reaction bottle3(2eq), tetrahydrofuran (5 fold by weight), after stirring to dissolve, Boc-beta-Ala-OSu (1eq) was added at room temperature and then stirred at room temperature until TLC monitored the reaction completion. Concentrating the reaction solution under reduced pressure, adjusting pH to 3-4 with dilute hydrochloric acid, and adding ethyl acetate (3)x5 times of weight portion), anhydrous Na2SO4Dried, filtered and concentrated to afford Boc-beta-Ala-Pro-OH as a red-brown oil (80% yield).
Of Boc-beta-Ala-Pro-OH1H NMR(d6-DMSO)：1.38(9H，s,3xCH3)；1.80-1.95(2H,m,CH2)；2.08-2.20(1H,m,CHH)；2.33-2.48(3H,m,CH2,CHH)；3.10-3.15(2H,m,CH2)；3.43-3.55(2H,m,CH2)；4.21(1H,dd,J 8.0,4.0Hz，CH)；6.64(1H,m,NH)；12.5(1H,br,OH)。
(2) Synthesis of H-dab (Boc) -NHBn
Adding Fmoc-dab (Boc) -OH (1eq) and dichloromethane (10-fold parts by weight based on Fmoc-dab (Boc) -OH) into a reaction flask, stirring, dissolving, sequentially adding DIC (1.2eq) and HOBT (1.2eq), stirring at room temperature for 30 minutes, and then adding benzylamine (1 eq); the reaction was run at room temperature until TLC monitored completion of the reaction. The reaction solution is washed by dilute hydrochloric acid, water and saturated saline water in sequence, dried by anhydrous sodium sulfate, filtered and concentrated to obtain Fmoc-dab (Boc) -NHBn, and the Fmoc-dab (Boc) -NHBn is directly used for the next reaction without purification (a small amount of analytically pure samples are obtained by a quick silica gel column for structural identification).
Of Fmoc-dab (Boc) -NHBn1H NMR(d6-DMSO)：1.38(9H，s,3xCH3)；1.66-1.87(2H,m,CH2)；2.93-3.02(2H,m,CH2)；3.35(2H,s,CH2)；4.03-4.08(1H,m,CH)；4.21-4.31(5H,m,CH,2xCH2)；6.75(1H,br,NH)；7.19-7.35(7H,m,CH)；7.40-7.44(2H,m,2xCH)；7.60(1H,br,NH)；7.74-7.75(2H,m,2xCH)；7.81-7.91(2H,m,2xCH)；8.37-8.39(1H,br,NH)。
Dissolving Fmoc-dab (Boc) -NHBn obtained in the previous step in methanol (10 times of weight parts), adding piperidine (5eq), reacting completely at room temperature, concentrating, adding the residue into dilute hydrochloric acid, extracting impurities with dichloromethane, adjusting the pH of the water phase to 10 with sodium carbonate, extracting with ethyl acetate, drying with anhydrous sodium sulfate, and concentrating to obtain the product H-dab (Boc) -NHBoc (purity is more than 95%, and yield in two steps is 70%).
Of H-L-dab (Boc) -NHBn1H NMR(d6-DMSO)：1.38-1.46(10H，m,3xCH3,CHH)；1.70-1.79(1H,m,CHH)；1.92(2H,br,NH2)；3.00-3.05(2H,m,CH2)；3.15-3.19(1H,m,CH)；4.28-4.30(2H,m,CH2)；6.78(1H,br,NH)；7.24-7.34(5H,m,5xCH)；8.37-8.39(1H,br,NH)。
(3) Synthesis of snake venom like peptide
Dichloromethane (20 times weight portion) and Boc-beta-Ala-Pro-OH (1.1eq) are added into a reaction bottle, DIC (1.2eq) is added after stirring and dissolving, H-dab (Boc) -NHBn (1.0eq) is added after stirring for 30 minutes, the reaction is stirred at room temperature, 5% sodium carbonate and 5% citric acid aqueous solution are respectively used for washing after TLC monitoring reaction is completed, anhydrous sodium sulfate is dried, and residue after filtration and concentration is crystallized by using mixed solvent (20 times weight portion) of ethyl acetate/petroleum ether with the mass ratio of 1:1 to obtain Boc-beta-Ala-Pro-dab (Boc) -NHBn (yield 88%, purity is more than 98%).
Of Boc-beta-Ala-Pro-dab (Boc) -NHBn1H NMR(d6-DMSO)：1.30-1.35(18H，m,6xCH3)；1.60-2.20(6H,m,3xCH2)；2.45-2.50(2H,m,CH2)；2.85-2.95(2H,m,CH2)；3.10-3.20(2H,m,CH2)；3.40-3.60(2H,m,CH2),4.20-4.40(4H,m,2xCH,CH2)；6.50-6.75(2H,m,2x NH),7.24-7.34(5H,m,5xCH)；7.8(1m,br,NH),8.20(1H,br,NH)。
And dissolving the Boc-beta-Ala-Pro-dab (Boc) -NHBn (1eq) obtained in the previous step in ethyl acetate (10 times of weight), dropwise adding HCl/ethyl acetate solution (4mol/L, 10 times of weight), monitoring by TLC (thin layer chromatography), filtering a solid precipitated in a reaction solution after the reaction is completed, rinsing (5 times of weight of ethyl acetate), and drying to obtain dihydrochloride of the snake venom tripeptide (yield 95% and purity more than 98%).
1H NMR(D2O):1.70-1.80(1H，m,CHH)；1.81-1.98(2H,m,CH2)；2.01-2.30(3H,m,CHH，CH2)；2.73-2.78(2H,m,CH2)；2.90-3.35(4H,m,2xCH2)；3.48-3.6(2H,m,，CH2)；4.30-4.40(4H，m,2xCH2)7.20-7.45(5H,m,5x CH)。
Example 2
(1) Synthesis of Boc-beta-Ala-Pro-OH
Adding L-Pro-OH (1.2eq) into a reaction flaskWater (5 times weight portions), NaHCO3(2eq), acetonitrile (5 fold by weight), after dissolution with stirring, Boc-beta-Ala-OSu (1eq, known compound) was added at room temperature and then stirred at room temperature until TLC monitored the reaction completion. Concentrating the reaction solution under reduced pressure, adjusting pH to 3-4 with dilute hydrochloric acid, extracting with ethyl acetate (3 × 5 times by weight), and extracting with anhydrous Na2SO4Dried, filtered and concentrated to afford Boc-beta-Ala-Pro-OH as a red-brown oil (75% yield).
(2) Synthesis of H-dab (Boc) -NHBn
Adding Fmoc-dab (Boc) -OH (1eq) and tetrahydrofuran (10-fold parts by weight based on Fmoc-dab (Boc) -OH) into a reaction flask, stirring, dissolving, sequentially adding DIC (1.2eq) and HOBT (1.2eq), stirring at room temperature for 30 minutes, and then adding benzylamine (1 eq); the reaction was run at room temperature until TLC monitored completion of the reaction. Washing the reaction solution with dilute hydrochloric acid, water and saturated saline successively, extracting with ethyl acetate, drying with anhydrous sodium sulfate, filtering and concentrating to obtain Fmoc-dab (Boc) -NHBn, and directly feeding the Fmoc-dab (Boc) -NHBn to the next step without purification for reaction.
Dissolving Fmoc-dab (Boc) -NHBn obtained in the previous step in dichloromethane (10eq), adding piperidine (5eq), reacting at room temperature completely, concentrating, adding the residue into dilute hydrochloric acid, extracting impurities with dichloromethane, adjusting the pH of the water phase to 10 with sodium carbonate, extracting with ethyl acetate, drying with anhydrous sodium sulfate, and concentrating to obtain the product H-dab (Boc) -NHBoc (purity is more than 95%, and yield in two steps is 65%).
(3) Synthesis of snake venom like peptide
Dichloromethane (20 times weight portion) and Boc-beta-Ala-Pro-OH (1.1eq) are added into a reaction bottle, DIC (1.2eq) is added after stirring and dissolving, H-dab (Boc) -NHBn (1.0eq) is added after stirring for 30 minutes, the reaction is stirred at room temperature, 5% sodium carbonate and 5% citric acid aqueous solution are respectively used for washing after TLC monitoring reaction is completed, anhydrous sodium sulfate is dried, and residue after filtration and concentration is crystallized by using mixed solvent (20 times weight portion) of ethyl acetate/petroleum ether with the mass ratio of 1:1 to obtain Boc-beta-Ala-Pro-dab (Boc) -NHBn (yield 89%, purity is more than 98%).
And dissolving the Boc-beta-Ala-Pro-dab (Boc) -NHBn (1eq) obtained in the previous step in ethyl acetate (10 times of weight), dropwise adding HCl/ethyl acetate solution (4mol/L, 10 times of weight), monitoring by TLC (thin layer chromatography), filtering a solid precipitated in a reaction solution after the reaction is completed, rinsing (5 times of weight of ethyl acetate), and drying to obtain dihydrochloride of the snake venom tripeptide (yield 93% and purity more than 98%).
Example 3
(1) Synthesis of Boc-beta-Ala-Pro-OH
Adding L-Pro-OH (1.2eq), water (5 times of the weight of the mixture) and NaHCO into a reaction bottle3(2eq), acetone (5 fold by weight), after dissolution with stirring, Boc-beta-Ala-OSu (1eq, N.C.) was added at room temperature, followed by stirring at room temperature until the reaction was complete as monitored by TLC. Concentrating the reaction solution under reduced pressure, adjusting pH to 3-4 with dilute hydrochloric acid, extracting with ethyl acetate (3 × 5 times by weight), and extracting with anhydrous Na2SO4Dried, filtered and concentrated to afford Boc-beta-Ala-Pro-OH as a red-brown oil (78% yield.
(2) Synthesis of H-dab (Boc) -NHBn
Adding Fmoc-dab (Boc) -OH (1eq) and dichloromethane (10 times of weight of the mixture based on Fmoc-dab (Boc) -OH) into a reaction bottle, stirring, dissolving, sequentially adding EDCI (1.2eq), stirring at room temperature for 30 minutes, and then adding benzylamine (1 eq); the reaction was run at room temperature until TLC monitored completion of the reaction. Washing the reaction solution with dilute hydrochloric acid, water and saturated saline successively, drying with anhydrous sodium sulfate, filtering and concentrating to obtain Fmoc-dab (Boc) -NHBn, and directly carrying out the next reaction without purification.
Dissolving Fmoc-dab (Boc) -NHBn obtained in the previous step in tetrahydrofuran (10 times of weight portion), then adding piperidine (5eq), after complete reaction at room temperature, concentrating, adding the residue into dilute hydrochloric acid, extracting impurities by ethyl acetate, adjusting pH of a water phase to 10 by using sodium carbonate, extracting by using ethyl acetate, drying by using anhydrous sodium sulfate, and concentrating to obtain a product H-dab (Boc) -NHBoc (purity is more than 95%, and yield in two steps is 68%).
(3) Synthesis of snake venom like peptide
Dichloromethane (20 times weight portion) and Boc-beta-Ala-Pro-OH (1.1eq) are added into a reaction bottle, DIC (1.2eq) is added after stirring and dissolving, H-dab (Boc) -NHBn (1.0eq) is added after stirring for 30 minutes, the reaction is stirred at room temperature, 5% sodium carbonate and 5% citric acid aqueous solution are respectively used for washing after TLC monitoring reaction is completed, anhydrous sodium sulfate is dried, and the residue after filtration and concentration is crystallized by using mixed solvent (20 times weight portion) with the mass ratio of ethyl acetate to petroleum ether being 1:1 to obtain Boc-beta-Ala-Pro-dab (Boc) -NHBn (yield is 88%, purity is more than 98%).
And dissolving the Boc-beta-Ala-Pro-dab (Boc) -NHBn (1eq) obtained in the previous step in ethyl acetate (10 times of weight), dropwise adding HCl/ethyl acetate solution (4mol/L, 10 times of weight), monitoring by TLC (thin layer chromatography), filtering a solid precipitated in a reaction solution after the reaction is completed, rinsing (5 times of weight of ethyl acetate), and drying to obtain dihydrochloride of the snake venom tripeptide (yield 95% and purity more than 98%).
Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.
Claims (10)
- A method for preparing Boc-beta-Ala-Pro-dab (Boc) -NHBn, comprising the steps of:the amino acid dehydration condensation reaction of Boc-beta-Ala-Pro-OH and H-dab (Boc) -NHBn is carried out to obtain:
- 2. the method of claim 1, wherein the Boc-beta-Ala-Pro-OH: the molar use ratio of H-dab (Boc) -NHBn is 1.0-1.5: 1.0, preferably 1.1: 1.0.
- 3. the method of claim 1, wherein the reaction conditions include a first condensing agent; further, the first condensing agent is selected from one or more of DIC, DCC and EDCI, preferably DIC;further, the first condensing agent: the molar use ratio of H-dab (Boc) -NHBn is 1.0-1.5: 1.0, preferably 1.2: 1.0;further, the solvent for the reaction is a first solvent, and the first solvent is selected from one or more of DCM, EtOAc and THF, preferably DCM;further, H-dab (Boc) -NHBn: the mass ratio of the first solvent is 1: 15-30; further 1: 20.
- 4. the process according to claim 1, wherein the reaction mixture is washed with an aqueous alkaline solution, an aqueous acidic solution and water, the solid-liquid separation is carried out after washing, and the solid is separated from the reaction mixture by ethyl acetate: the mass ratio of petroleum ether is 0.8-1.2: crystallizing and purifying by using 0.8-1.2 of organic solvent;further, the alkaline aqueous solution is a sodium carbonate aqueous solution with the mass fraction of 2-10%, preferably a sodium carbonate aqueous solution with the mass fraction of 5%;further, the acidic aqueous solution is 2-10% by mass of citric acid aqueous solution, preferably 5% by mass of citric acid aqueous solution;further, the mass usage amount of the organic solvent is 15-25 times of the mass of H-dab (Boc) -NHBn, and is further 20 times;further, ethyl acetate: the mass ratio of the petroleum ether is 1: 1.
- 5. the method of claim 1, wherein the H-dab (Boc) -NHBn is obtained by dehydrating condensation of Fmoc-dab (Boc) -OH with benzylamine and removal of Fmoc group:
further, Fmoc-dab (boc) -OH: the molar use ratio of benzylamine is 0.8-1.2: 0.8 to 1.2, preferably 1: 1. - 6. the method according to claim 5, wherein the conditions for dehydration condensation reaction of Fmoc-dab (Boc) -OH with benzylamine amino acid include a second condensing agent; further, the second condensing agent is selected from one or more of DIC, DCC, EDCI, HOBT, CDI, HBTU and HATU, preferably DIC and/or HOBT, or EDCI; more preferably DIC and HOBT, DIC: the HOBT molar use ratio is 1: 1;further, the second condensing agent: the molar use ratio of Fmoc-dab (Boc) -OH is 1.0-3.0: 1.0, preferably 1.2 to 2.4: 1.0.
- 7. the preparation method according to claim 5, wherein the solvent for the dehydration condensation reaction of Fmoc-dab (Boc) -OH with benzylamine is a second solvent selected from one or more of DCM, THF and acetonitrile, preferably DCM and/or THF, more preferably DCM;further, Fmoc-dab (boc) -OH: the mass ratio of the second solvent is 1: 5-15, preferably 1: 10.
- 8. the method of claim 1, wherein the Boc-beta-Ala-Pro-OH is obtained by reacting Boc-beta-Ala-OSu with L-Pro-OH by:
further, Boc-beta-Ala-OSu: the molar use ratio of L-Pro-OH is 1.0: 1.0-1.5, preferably 1; 1.2;further, the conditions for the reaction of Boc-beta-Ala-OSu with L-Pro-OH include a base; further, the base is selected from NaHCO3、KHCO3、Na2CO3、K2CO3Preferably NaHCO3；Further, Boc-beta-Ala-OSu: the molar ratio of the alkali is 1.0: 1.5 to 3, preferably 1.0: 2.0.further, the solvent for the reaction of Boc-beta-Ala-OSu with L-Pro-OH was water: the mass ratio of the organic solvent is 0.8-1.2: 0.8-1.2, preferably water: the mass ratio of the organic solvent is 1:1, wherein the organic solvent is selected from one or more of tetrahydrofuran, acetonitrile, acetone, methanol, ethanol and DMF, preferably one or more of tetrahydrofuran, acetonitrile and acetone, and more preferably tetrahydrofuran;further, Boc-beta-Ala-OSu: the mass ratio of the mixed solvent is 1: 5-15, preferably 1: 10. - 9. the preparation method of snake venom tripeptide dihydrochloride is characterized in that the snake venom tripeptide dihydrochloride is obtained by removing Boc group through Boc-beta-Ala-Pro-dab (Boc) -NHBn;Boc-beta-Ala-Pro-dab (Boc) -NHBn prepared by the method of any one of claims 1 to 8;further, mixing Boc-beta-Ala-Pro-dab (Boc) -NHBn with ethyl acetate, adding HCl ethyl acetate solution into the solution in batches, and after the reaction is finished, carrying out solid-liquid separation to obtain a solid matter which is snake venom tripeptide dihydrochloride;further, Boc-beta-Ala-Pro-dab (Boc) -NHBn: the mass ratio of ethyl acetate is 1: 5-15, preferably 1: 10; the concentration of HCl in ethyl acetate is 3-5 mol/L, preferably 4mol/L, Boc-beta-Ala-Pro-dab (Boc) -NHBn: the mass usage ratio of the HCl ethyl acetate solution is 1: 5-15, preferably 1: 10.
- 10. use of a snake venom-like tripeptide dihydrochloride for preparing a product related to a snake venom-like tripeptide, wherein the snake venom-like tripeptide dihydrochloride is prepared by the preparation method according to claim 9.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011097855.2A CN114369139B (en) | 2020-10-14 | 2020-10-14 | Preparation method of snake venom tripeptide |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011097855.2A CN114369139B (en) | 2020-10-14 | 2020-10-14 | Preparation method of snake venom tripeptide |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN114369139A true CN114369139A (en) | 2022-04-19 |
| CN114369139B CN114369139B (en) | 2024-04-19 |
Family
ID=81138411
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202011097855.2A Active CN114369139B (en) | 2020-10-14 | 2020-10-14 | Preparation method of snake venom tripeptide |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114369139B (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116375601A (en) * | 2023-06-05 | 2023-07-04 | 成都泰莱康科技有限公司 | Anti-melanoma compound and preparation method and application thereof |
| CN117486968A (en) * | 2024-01-03 | 2024-02-02 | 深圳创元生物医药科技有限公司 | Preparation method of snake venom peptide |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101061134A (en) * | 2004-11-02 | 2007-10-24 | 彭特法姆股份公司 | Novel topical application agents against mimic and age-related wrinkles |
| US20100215726A1 (en) * | 2009-02-25 | 2010-08-26 | Peter Thomas Roth | Compositions for Diminishing the Appearance of Wrinkles |
| RU2551276C1 (en) * | 2014-02-03 | 2015-05-20 | Закрытое Акционерное Общество "Фарм-Синтез" | Method for producing tripeptide diacetate |
| CN107857797A (en) * | 2017-12-07 | 2018-03-30 | 陕西慧康生物科技有限责任公司 | The liquid-phase fragment synthetic method of one species snake venom tripeptides |
| CN107936108A (en) * | 2017-12-05 | 2018-04-20 | 陕西慧康生物科技有限责任公司 | The liquid-phase synthesis process of one species snake venom tripeptides |
-
2020
- 2020-10-14 CN CN202011097855.2A patent/CN114369139B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101061134A (en) * | 2004-11-02 | 2007-10-24 | 彭特法姆股份公司 | Novel topical application agents against mimic and age-related wrinkles |
| US20100215726A1 (en) * | 2009-02-25 | 2010-08-26 | Peter Thomas Roth | Compositions for Diminishing the Appearance of Wrinkles |
| RU2551276C1 (en) * | 2014-02-03 | 2015-05-20 | Закрытое Акционерное Общество "Фарм-Синтез" | Method for producing tripeptide diacetate |
| CN107936108A (en) * | 2017-12-05 | 2018-04-20 | 陕西慧康生物科技有限责任公司 | The liquid-phase synthesis process of one species snake venom tripeptides |
| CN107857797A (en) * | 2017-12-07 | 2018-03-30 | 陕西慧康生物科技有限责任公司 | The liquid-phase fragment synthetic method of one species snake venom tripeptides |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116375601A (en) * | 2023-06-05 | 2023-07-04 | 成都泰莱康科技有限公司 | Anti-melanoma compound and preparation method and application thereof |
| CN116375601B (en) * | 2023-06-05 | 2023-08-04 | 成都泰莱康科技有限公司 | Anti-melanoma compound and preparation method and application thereof |
| CN117486968A (en) * | 2024-01-03 | 2024-02-02 | 深圳创元生物医药科技有限公司 | Preparation method of snake venom peptide |
| CN117486968B (en) * | 2024-01-03 | 2024-03-29 | 深圳创元生物医药科技有限公司 | Preparation method of snake venom peptide |
Also Published As
| Publication number | Publication date |
|---|---|
| CN114369139B (en) | 2024-04-19 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN114369139B (en) | Preparation method of snake venom tripeptide | |
| CN112430253A (en) | Liquid phase synthesis method of snake venom-like tripeptide | |
| CN110950926B (en) | Liquid phase synthesis method of snake venom-like tripeptide | |
| CN111978369B (en) | Method for preparing polypeptide | |
| CN107474107A (en) | GLYX 13 preparation method and the compound for preparing GLYX 13 | |
| CN114213503A (en) | Synthesis method of snake venom-like tripeptide | |
| CN109879936B (en) | Method for synthesizing palmitoyl hexapeptide by using micro-channel modular reaction device | |
| CN113667007A (en) | Liquid-phase preparation method of side chain of Somaloutide | |
| CN113045623A (en) | Liquid phase synthesis method of snake venom peptide SYN-AKE | |
| CN115197150A (en) | Preparation method of L-carnosine | |
| JP7278775B2 (en) | Method for producing long-chain compounds | |
| CN110183532B (en) | Process method for synthesizing bivalirudin protected pentapeptide fragments by mass high-efficiency liquid phase method | |
| CN111690037A (en) | Method for synthesizing GHK acetate | |
| CN100363380C (en) | Liquid phase synthetic method for endomorphine -1 and endomorphine -2 | |
| CN109776505B (en) | Preparation method of avanafil | |
| CN117486968B (en) | Preparation method of snake venom peptide | |
| CN111269261A (en) | Liquid phase total synthesis method of TSBP auxiliary group and group-assisted enkephalin and derivative thereof | |
| CN111285921A (en) | BDK auxiliary group and liquid phase total synthesis method of procapsipeptide and analogue based on BDK auxiliary group | |
| CN115368221B (en) | Small molecule carrier, preparation method thereof and application thereof in polypeptide synthesis | |
| CN116606233A (en) | Preparation method of S-trityl-L-cysteamine | |
| CN102146065A (en) | Method for preparing high-purity diketopiperazine cyclopeptide | |
| CN115043903B (en) | Liquid phase synthesis method of snake venom-like tripeptide | |
| CN115636866A (en) | Method for synthesizing bridged ring compound and intermediate thereof | |
| CN116332810B (en) | Preparation method of Fmoc-His (Trt) -Abe-OH | |
| CN109251150B (en) | Preparation method of 2, 3-diamino methyl propionate |
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 | ||
| TA01 | Transfer of patent application right |
Effective date of registration: 20230426 Address after: 611130 No. 1-2, building 5, No. 288, north section of Rongtai Avenue, Chengdu cross strait science and Technology Industrial Development Park, Wenjiang District, Chengdu, Sichuan Applicant after: Chengdu Tailaikang Technology Co.,Ltd. Address before: 611130 No. 1-2, building 5, No. 288, north section of Rongtai Avenue, Chengdu cross strait science and Technology Industrial Development Park, Wenjiang District, Chengdu, Sichuan Applicant before: Chengdu Tailaikang Technology Co.,Ltd. Applicant before: CHENGDU YONTINOTECH CO.,LTD. |
|
| TA01 | Transfer of patent application right | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |