CN105439883A - Preparation method and application of D-dencichine - Google Patents
Preparation method and application of D-dencichine Download PDFInfo
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- 238000002360 preparation method Methods 0.000 title claims abstract description 32
- 206010043554 thrombocytopenia Diseases 0.000 claims abstract description 17
- DCXYFEDJOCDNAF-UWTATZPHSA-N D-Asparagine Chemical compound OC(=O)[C@H](N)CC(N)=O DCXYFEDJOCDNAF-UWTATZPHSA-N 0.000 claims abstract description 16
- 239000003814 drug Substances 0.000 claims abstract description 15
- 230000000694 effects Effects 0.000 claims abstract description 14
- 125000003088 (fluoren-9-ylmethoxy)carbonyl group Chemical group 0.000 claims abstract description 10
- 150000007530 organic bases Chemical class 0.000 claims abstract description 8
- 238000006482 condensation reaction Methods 0.000 claims abstract description 4
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 108
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 60
- 238000006243 chemical reaction Methods 0.000 claims description 59
- 239000007787 solid Substances 0.000 claims description 51
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 44
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims description 42
- 238000000967 suction filtration Methods 0.000 claims description 37
- MUBZPKHOEPUJKR-UHFFFAOYSA-N Oxalic acid Chemical compound OC(=O)C(O)=O MUBZPKHOEPUJKR-UHFFFAOYSA-N 0.000 claims description 33
- UHOVQNZJYSORNB-UHFFFAOYSA-N Benzene Chemical compound C1=CC=CC=C1 UHOVQNZJYSORNB-UHFFFAOYSA-N 0.000 claims description 30
- 239000011541 reaction mixture Substances 0.000 claims description 29
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 26
- WMFOQBRAJBCJND-UHFFFAOYSA-M Lithium hydroxide Chemical compound [Li+].[OH-] WMFOQBRAJBCJND-UHFFFAOYSA-M 0.000 claims description 24
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 16
- JUJWROOIHBZHMG-UHFFFAOYSA-N Pyridine Chemical compound C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 claims description 16
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 claims description 16
- 235000006408 oxalic acid Nutrition 0.000 claims description 11
- 229910052700 potassium Inorganic materials 0.000 claims description 11
- 239000011591 potassium Substances 0.000 claims description 11
- -1 potassium ester Chemical class 0.000 claims description 11
- SNOFKGQEUYMNCE-UHFFFAOYSA-N 2,2,2-trifluoro-1-$l^{1}-oxidanylethanone Chemical compound [O]C(=O)C(F)(F)F SNOFKGQEUYMNCE-UHFFFAOYSA-N 0.000 claims description 10
- PNDPGZBMCMUPRI-UHFFFAOYSA-N iodine Chemical compound II PNDPGZBMCMUPRI-UHFFFAOYSA-N 0.000 claims description 10
- 229910052740 iodine Inorganic materials 0.000 claims description 10
- 239000011630 iodine Substances 0.000 claims description 10
- 229940079593 drug Drugs 0.000 claims description 8
- FUKUFMFMCZIRNT-UHFFFAOYSA-N hydron;methanol;chloride Chemical compound Cl.OC FUKUFMFMCZIRNT-UHFFFAOYSA-N 0.000 claims description 8
- UMJSCPRVCHMLSP-UHFFFAOYSA-N pyridine Natural products COC1=CC=CN=C1 UMJSCPRVCHMLSP-UHFFFAOYSA-N 0.000 claims description 8
- 229910000030 sodium bicarbonate Inorganic materials 0.000 claims description 8
- 235000017557 sodium bicarbonate Nutrition 0.000 claims description 8
- HPNMFZURTQLUMO-UHFFFAOYSA-N diethylamine Chemical compound CCNCC HPNMFZURTQLUMO-UHFFFAOYSA-N 0.000 claims description 6
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 claims description 5
- 238000005406 washing Methods 0.000 claims description 5
- 239000002253 acid Substances 0.000 claims description 4
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims description 4
- 239000008194 pharmaceutical composition Substances 0.000 claims description 4
- UIIMBOGNXHQVGW-DEQYMQKBSA-M Sodium bicarbonate-14C Chemical compound [Na+].O[14C]([O-])=O UIIMBOGNXHQVGW-DEQYMQKBSA-M 0.000 claims description 2
- 238000002156 mixing Methods 0.000 claims description 2
- 238000010979 pH adjustment Methods 0.000 claims description 2
- 150000003053 piperidines Chemical class 0.000 claims description 2
- 238000005057 refrigeration Methods 0.000 claims description 2
- 238000010025 steaming Methods 0.000 claims description 2
- 102000003815 Interleukin-11 Human genes 0.000 abstract description 11
- 108090000177 Interleukin-11 Proteins 0.000 abstract description 11
- 229940074383 interleukin-11 Drugs 0.000 abstract description 11
- 238000000034 method Methods 0.000 abstract description 5
- 150000001875 compounds Chemical class 0.000 abstract description 3
- 230000009471 action Effects 0.000 abstract description 2
- CPKISUMKCULUNR-UHFFFAOYSA-N 2-methoxy-2-oxoacetic acid Chemical compound COC(=O)C(O)=O CPKISUMKCULUNR-UHFFFAOYSA-N 0.000 abstract 1
- 229930182846 D-asparagine Natural products 0.000 abstract 1
- 125000003277 amino group Chemical group 0.000 abstract 1
- 238000006731 degradation reaction Methods 0.000 abstract 1
- 238000003756 stirring Methods 0.000 description 61
- 239000000243 solution Substances 0.000 description 38
- 230000015572 biosynthetic process Effects 0.000 description 25
- 238000003786 synthesis reaction Methods 0.000 description 25
- 190000008236 carboplatin Chemical compound 0.000 description 20
- 229960004562 carboplatin Drugs 0.000 description 20
- 239000012153 distilled water Substances 0.000 description 19
- 238000002347 injection Methods 0.000 description 16
- 239000007924 injection Substances 0.000 description 16
- 210000001772 blood platelet Anatomy 0.000 description 14
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 12
- 239000007864 aqueous solution Substances 0.000 description 12
- 241001465754 Metazoa Species 0.000 description 9
- 229960004756 ethanol Drugs 0.000 description 7
- PEZNEXFPRSOYPL-UHFFFAOYSA-N (bis(trifluoroacetoxy)iodo)benzene Chemical compound FC(F)(F)C(=O)OI(OC(=O)C(F)(F)F)C1=CC=CC=C1 PEZNEXFPRSOYPL-UHFFFAOYSA-N 0.000 description 6
- 238000001644 13C nuclear magnetic resonance spectroscopy Methods 0.000 description 6
- 238000005160 1H NMR spectroscopy Methods 0.000 description 6
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 6
- 241000700159 Rattus Species 0.000 description 6
- 210000004369 blood Anatomy 0.000 description 6
- 239000008280 blood Substances 0.000 description 6
- 229960000935 dehydrated alcohol Drugs 0.000 description 6
- 238000001514 detection method Methods 0.000 description 6
- 239000007788 liquid Substances 0.000 description 6
- 238000012544 monitoring process Methods 0.000 description 6
- 239000002994 raw material Substances 0.000 description 6
- 230000004044 response Effects 0.000 description 6
- 238000009534 blood test Methods 0.000 description 5
- 238000010253 intravenous injection Methods 0.000 description 5
- 239000002904 solvent Substances 0.000 description 5
- 241000699666 Mus <mouse, genus> Species 0.000 description 4
- NEEQFPMRODQIKX-REOHCLBHSA-N N(3)-oxalyl-L-2,3-diaminopropionic acid Chemical compound OC(=O)[C@@H](N)CNC(=O)C(O)=O NEEQFPMRODQIKX-REOHCLBHSA-N 0.000 description 4
- 229940000406 drug candidate Drugs 0.000 description 4
- 239000002504 physiological saline solution Substances 0.000 description 4
- 238000012360 testing method Methods 0.000 description 4
- 244000131316 Panax pseudoginseng Species 0.000 description 3
- 235000003181 Panax pseudoginseng Nutrition 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 206010014080 Ecchymosis Diseases 0.000 description 2
- 208000032843 Hemorrhage Diseases 0.000 description 2
- 238000010241 blood sampling Methods 0.000 description 2
- 230000037396 body weight Effects 0.000 description 2
- 210000001185 bone marrow Anatomy 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 210000003677 hemocyte Anatomy 0.000 description 2
- 229940000351 hemocyte Drugs 0.000 description 2
- 230000023597 hemostasis Effects 0.000 description 2
- 230000003284 homeostatic effect Effects 0.000 description 2
- 238000010172 mouse model Methods 0.000 description 2
- 210000004279 orbit Anatomy 0.000 description 2
- 210000005259 peripheral blood Anatomy 0.000 description 2
- 239000011886 peripheral blood Substances 0.000 description 2
- 239000013641 positive control Substances 0.000 description 2
- SCVFZCLFOSHCOH-UHFFFAOYSA-M potassium acetate Chemical compound [K+].CC([O-])=O SCVFZCLFOSHCOH-UHFFFAOYSA-M 0.000 description 2
- 230000000630 rising effect Effects 0.000 description 2
- XDBMTQVSHNQIFU-UHFFFAOYSA-N 2-(2-ethoxy-2-oxo-1-phenylethyl)-1,3-thiazolidin-3-ium-4-carboxylate Chemical compound C=1C=CC=CC=1C(C(=O)OCC)C1NC(C(O)=O)CS1 XDBMTQVSHNQIFU-UHFFFAOYSA-N 0.000 description 1
- 206010002091 Anaesthesia Diseases 0.000 description 1
- 206010065553 Bone marrow failure Diseases 0.000 description 1
- 206010051625 Conjunctival hyperaemia Diseases 0.000 description 1
- 102000004127 Cytokines Human genes 0.000 description 1
- 108090000695 Cytokines Proteins 0.000 description 1
- 206010015150 Erythema Diseases 0.000 description 1
- 101001010568 Homo sapiens Interleukin-11 Proteins 0.000 description 1
- 229930010555 Inosine Natural products 0.000 description 1
- UGQMRVRMYYASKQ-KQYNXXCUSA-N Inosine Chemical compound O[C@@H]1[C@H](O)[C@@H](CO)O[C@H]1N1C2=NC=NC(O)=C2N=C1 UGQMRVRMYYASKQ-KQYNXXCUSA-N 0.000 description 1
- 241000608867 Leucogenes Species 0.000 description 1
- LOMVENUNSWAXEN-UHFFFAOYSA-N Methyl oxalate Chemical compound COC(=O)C(=O)OC LOMVENUNSWAXEN-UHFFFAOYSA-N 0.000 description 1
- 206010061298 Mucosal haemorrhage Diseases 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 206010029350 Neurotoxicity Diseases 0.000 description 1
- 206010030113 Oedema Diseases 0.000 description 1
- 208000002193 Pain Diseases 0.000 description 1
- 241000208343 Panax Species 0.000 description 1
- 206010068676 Pneumoretroperitoneum Diseases 0.000 description 1
- 208000005727 Retropneumoperitoneum Diseases 0.000 description 1
- NSOXQYCFHDMMGV-UHFFFAOYSA-N Tetrakis(2-hydroxypropyl)ethylenediamine Chemical compound CC(O)CN(CC(C)O)CCN(CC(C)O)CC(C)O NSOXQYCFHDMMGV-UHFFFAOYSA-N 0.000 description 1
- 206010044221 Toxic encephalopathy Diseases 0.000 description 1
- 206010046910 Vaginal haemorrhage Diseases 0.000 description 1
- 230000037005 anaesthesia Effects 0.000 description 1
- 230000000118 anti-neoplastic effect Effects 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 210000003169 central nervous system Anatomy 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 230000000973 chemotherapeutic effect Effects 0.000 description 1
- RNFNDJAIBTYOQL-UHFFFAOYSA-N chloral hydrate Chemical compound OC(O)C(Cl)(Cl)Cl RNFNDJAIBTYOQL-UHFFFAOYSA-N 0.000 description 1
- 229960002327 chloral hydrate Drugs 0.000 description 1
- 239000013078 crystal Substances 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 238000001647 drug administration Methods 0.000 description 1
- 208000001780 epistaxis Diseases 0.000 description 1
- 230000003203 everyday effect Effects 0.000 description 1
- 230000002489 hematologic effect Effects 0.000 description 1
- 230000002607 hemopoietic effect Effects 0.000 description 1
- 102000049885 human IL11 Human genes 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 230000001965 increasing effect Effects 0.000 description 1
- 229960003786 inosine Drugs 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- 230000000388 leucogen effect Effects 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000010534 mechanism of action Effects 0.000 description 1
- 239000000178 monomer Substances 0.000 description 1
- 208000011309 nasal bleeding Diseases 0.000 description 1
- 230000002887 neurotoxic effect Effects 0.000 description 1
- 231100000228 neurotoxicity Toxicity 0.000 description 1
- 230000007135 neurotoxicity Effects 0.000 description 1
- 230000036407 pain Effects 0.000 description 1
- 230000007170 pathology Effects 0.000 description 1
- 235000011056 potassium acetate Nutrition 0.000 description 1
- 238000011552 rat model Methods 0.000 description 1
- 238000005215 recombination Methods 0.000 description 1
- 230000006798 recombination Effects 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 230000000452 restraining effect Effects 0.000 description 1
- 208000009146 rhinoscleroma Diseases 0.000 description 1
- 210000000952 spleen Anatomy 0.000 description 1
- 238000010189 synthetic method Methods 0.000 description 1
- 230000003582 thrombocytopenic effect Effects 0.000 description 1
- 210000000689 upper leg Anatomy 0.000 description 1
- 238000010792 warming Methods 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C269/00—Preparation of derivatives of carbamic acid, i.e. compounds containing any of the groups, the nitrogen atom not being part of nitro or nitroso groups
- C07C269/04—Preparation of derivatives of carbamic acid, i.e. compounds containing any of the groups, the nitrogen atom not being part of nitro or nitroso groups from amines with formation of carbamate groups
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07B—GENERAL METHODS OF ORGANIC CHEMISTRY; APPARATUS THEREFOR
- C07B57/00—Separation of optically-active compounds
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C227/00—Preparation of compounds containing amino and carboxyl groups bound to the same carbon skeleton
- C07C227/14—Preparation of compounds containing amino and carboxyl groups bound to the same carbon skeleton from compounds containing already amino and carboxyl groups or derivatives thereof
- C07C227/18—Preparation of compounds containing amino and carboxyl groups bound to the same carbon skeleton from compounds containing already amino and carboxyl groups or derivatives thereof by reactions involving amino or carboxyl groups, e.g. hydrolysis of esters or amides, by formation of halides, salts or esters
- C07C227/20—Preparation of compounds containing amino and carboxyl groups bound to the same carbon skeleton from compounds containing already amino and carboxyl groups or derivatives thereof by reactions involving amino or carboxyl groups, e.g. hydrolysis of esters or amides, by formation of halides, salts or esters by hydrolysis of N-acylated amino-acids or derivatives thereof, e.g. hydrolysis of carbamates
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C231/00—Preparation of carboxylic acid amides
- C07C231/02—Preparation of carboxylic acid amides from carboxylic acids or from esters, anhydrides, or halides thereof by reaction with ammonia or amines
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C269/00—Preparation of derivatives of carbamic acid, i.e. compounds containing any of the groups, the nitrogen atom not being part of nitro or nitroso groups
- C07C269/06—Preparation of derivatives of carbamic acid, i.e. compounds containing any of the groups, the nitrogen atom not being part of nitro or nitroso groups by reactions not involving the formation of carbamate groups
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07B—GENERAL METHODS OF ORGANIC CHEMISTRY; APPARATUS THEREFOR
- C07B2200/00—Indexing scheme relating to specific properties of organic compounds
- C07B2200/07—Optical isomers
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- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention discloses a preparation method and application of D-dencichine. The preparation method comprises the following steps: carrying out Fmoc protection on the amino group of D-asparagine so as to obtain a first intermediate; subjecting the first intermediate to Hoffman degradation reaction so as to obtain a second intermediate; subjecting the second intermediate to removal of Fmoc protection under the action of organic base so as to obtain a third intermediate; and subjecting the third intermediate and monomethyl oxalate to condensation reaction under a highly basic condition so as to obtain D-dencichine. D-dencichine is prepared by using the high-efficiency safe preparation method; the preparation method is simple and has high yield; the compound D-dencichine prepared by using the method has good treatment effect on thrombocytopenia and the effect is better than the effect of a clinical medicine interleukin-11, so D-dencichine can be used as a candidate medicine for treating thrombocytopenia.
Description
Technical field
The present invention relates to pharmaceutical chemistry and therapeutics field, particularly relate to the preparation method and its usage of a kind of D-dencichine (i.e. β-N-ethanedioyl-D-α, β-diaminopropionic acid).
Background technology
Thrombocytopenia refers to that platelet count is lower than the illness caused by normal range, thrombocytopenia may come from thrombocyte and produce not enough, spleen is to hematoblastic detention, platelet destruction or utilization increase and are diluted, serious thrombopenia no matter caused by which kind of reason, all can cause typically hemorrhage: multiple ecchymosis, is most commonly in shank; Or there is little being dispersed in property ecchymosis at the position by microtrauma; Mucosal bleeding, nasal bleeding, gi tract and urogenital tract and vaginal hemorrhage, and Post operation is bled profusely, gi tract are bled profusely and central nervous system internal hemorrhage can threat to life.
Treatment thrombopenia drug main common clinically will have leucogen sheet, inosine injection, recombination human interleukins-11 etc. at present.But these medicines, in clinical application or action effect not obvious or occur all unsatisfactory side effects, just there is the side effects such as such as oedema, injection place pain, redness, scleroma, conjunctival congestion as interleukin-11.Therefore finding evident in efficacy, safe and reliable treatment thrombopenia disease drug is the problem that medical personal is worth studying for a long period of time.
Araliaceae Panax rare traditional Chinese medicine pseudo-ginseng has the title of " the refreshing medicine of hemostasis ", is traumatology key medicine since ancient times, and principal monomer styptic activity material contained in pseudo-ginseng is L-dencichine.L-dencichine, when a large amount of use, will show neurotoxicity, and when using on a small quantity, then not show Neurotoxic effect and demonstrate powerful hemostasis and thrombocyte increasing action, and use 1-2mg fully can show result for the treatment of.Pseudo-ginseng have two chiral isomers, but from natural phant be separated obtain be L configuration.
Summary of the invention
In view of this, the object of the invention is to the preparation method and its usage proposing a kind of D-dencichine (i.e. β-N-ethanedioyl-D-α, β-diaminopropionic acid), to prepare D-dencichine efficiently.
The preparation method of D-dencichine provided by the invention, comprises the following steps:
1) Fmoc protection is carried out to the amino of D-Asn, obtain the first intermediate;
2) Hoffman DeR is carried out to described first intermediate, obtain the second intermediate;
3) under the effect of organic bases, described second intermediate is sloughed Fmoc protection, obtain the 3rd intermediate;
4) under basic conditions, described 3rd intermediate and oxalic acid list potassium ester are carried out condensation reaction, obtains described D-dencichine.
In some embodiments of the invention, 9-fluorene methyl-N-succinimidyl carbonate is adopted to carry out Fmoc protection to the amino of D-Asn.
In some embodiments of the invention, adopt [two (trifluoroacetyl oxygen base) iodine] benzene to carry out Hoffman DeR to described first intermediate, obtain the second intermediate.
In some embodiments of the invention, described organic bases is selected from diethylamine or piperidines.
In some embodiments of the invention, described highly basic is selected from lithium hydroxide.
In some embodiments of the invention, the preparation method of described first intermediate comprises the following steps:
Sodium bicarbonate is joined in D-Asn, 9-fluorene methyl-N-succinimidyl carbonate is dissolved in acetone;
The acetone soln of described 9-fluorene methyl-N-succinimidyl carbonate is slowly added dropwise in the mixing solutions of sodium bicarbonate, D-Asn, reacts at 20-30 DEG C;
Adopt acid solution to regulate the pH value of reaction mixture to 2-4, then this reaction mixture of suction filtration, obtains white solid, is described first intermediate.
In some embodiments of the invention, the preparation method of described second intermediate comprises the following steps:
[two (trifluoroacetyl oxygen base) iodine] benzene is dissolved in acetonitrile, described first intermediate is dissolved in DMF, the described DMF being dissolved with the first intermediate is joined in acetonitrile solution;
Continue to add water to acetonitrile solution, and then continue to add pyridine to acetonitrile solution, react at 20-30 DEG C;
After reaction terminates, steaming desolventizes, and then adds water, adopts acid solution to regulate the pH value of reaction mixture to 1-3;
Then, adopt ether to extract, water layer adopts lye pH adjustment value to 5-7, and after refrigeration, suction filtration obtains white solid, is described second intermediate.
In some embodiments of the invention, the preparation method of described 3rd intermediate comprises the following steps:
Described second intermediate is dissolved in ethanol, then continues to add organic bases, react at 20-30 DEG C;
After reaction terminates, concentrating under reduced pressure reaction mixture, adopts washing with alcohol;
Add methyl alcohol and methanol hydrochloride solution to the reaction mixture after washing with alcohol, after adularescent solid is separated out, obtain white solid through suction filtration, methanol wash, be described 3rd intermediate.
The present invention also provides a kind of pharmaceutical composition, and it comprises the D-dencichine adopting the preparation method of above-mentioned D-dencichine to obtain.
The present invention also provides the purposes of a kind of aforementioned pharmaceutical compositions in preparation treatment thrombopenia disease drug.
As can be seen from the above, the present invention adopts the preparation method of highly effective and safe to synthesize and obtains D-dencichine, preparation technology is simple, yield is higher, Compound D-the dencichine prepared by method provided by the invention has treatment thrombocytopenia effect preferably, be better than clinical application interleukin-11, the drug candidate for the treatment of thrombocytopenia can be become.
Embodiment
For making the object, technical solutions and advantages of the present invention clearly understand, below in conjunction with specific embodiment, the present invention is described in more detail.
The preparation method of D-dencichine provided by the invention comprises the following steps:
1) Fmoc protection is carried out to the amino of D-Asn, obtain the first intermediate S-D-1;
2) Hoffman DeR is carried out to described first intermediate S-D-1, obtain the second intermediate S-D-2;
3) under the effect of organic bases, described second intermediate S-D-2 is sloughed Fmoc protection, obtain the 3rd intermediate S-D-3;
4) under basic conditions, described 3rd intermediate S-D-2 and oxalic acid list potassium ester are carried out condensation reaction, obtains target product D-dencichine.
The synthetic route of D-dencichine is as follows:
The synthetic method of embodiment 1D-dencichine
1) synthesis of the first intermediate S-D-1:
Get D-Asn one water thing (8.7g, 58mmol) in round-bottomed flask, add distilled water 300mL, add sodium bicarbonate (10.9g, 130mmol) subsequently and fully stir; Getting 9-fluorene methyl-N-succinimidyl carbonate (20g, 60mmol) is dissolved in 150mL acetone, is slowly added dropwise in above-mentioned round-bottomed flask, stirring reaction 4h at 25 DEG C by the acetone soln of described 9-fluorene methyl-N-succinimidyl carbonate; Concentrated hydrochloric acid is adopted to regulate the pH value to 3 of reaction mixture, then this reaction mixture of suction filtration, obtain 19.5g white solid, be the first intermediate S-D-1, yield 95%.
2) synthesis of the second intermediate S-D-2:
Get (24.9g, 56mmol) [two (trifluoroacetyl oxygen base) iodine] benzene (PIFA) in round-bottomed flask, add 58mL acetonitrile and dissolve; Separately get (19.2g, 54.2mmol) S-D-1, be dissolved in 115mLDMF, and be slowly added dropwise in described round-bottomed flask, vigorous stirring is clarified to reaction solution; Slowly drip subsequently in distilled water 58mL to described round-bottomed flask, after stirring 20min, slowly drip in pyridine 4.6mL to described round-bottomed flask, and at 26 DEG C stirring reaction 60h.Monitor raw material reaction completely by TLC, steam after desolventizing, add distilled water 380mL and stir, the pH value to 2 of 6NHCl solution regulator solution; Adopt ether to extract again, water layer 4NNaOH solution adjust pH is that after 2h placed by 6,4 DEG C of refrigerators, suction filtration obtains 13.3g white solid, is the second intermediate S-D-2, yield 75%.
3) synthesis of the 3rd intermediate S-D-3:
Get S-D-2 (13.0g, 40mmol) in round-bottomed flask, add dehydrated alcohol 1000mL, subsequently to dripping diethylamine 200mL in reaction flask, reacting at 27 DEG C and spending the night.TLC detection reaction completely after, by repeatedly concentrating under reduced pressure ethanol, remove residual quadrol.Add anhydrous methanol 100mL stirring and dissolving, drip 5mL methanol hydrochloride solution, after stirring reaction 4h, adularescent solid is separated out, and suction filtration, methanol wash column, obtains 5.0g white solid S-D-3, yield 90%.
4) synthesis of target product D-dencichine:
Get S-D-3 (5.3g, 36mmol) in round-bottomed flask, add 26.5mL50% methanol aqueous solution and stir, in ice bath downhill reaction bottle, add lithium hydroxide one water thing (2.3g, 51mmol) subsequently, stirring reaction 1h; After question response liquid dissolves, slowly drip the 32mL50% methanol aqueous solution of oxalic acid list potassium ester (9.7g, 65mmol), and react 24h under room temperature.After TLC monitoring reacts completely, suction filtration reaction mixture, is dissolved in frozen water by the white solid obtained, and adopts 2NHCl adjust pH to be 2.1 ~ 2.2, make again to separate out white solid, by it after 2h placed by 4 DEG C of refrigerators, suction filtration, carries out frozen water successively and washes, methanol wash column, after acetone is washed, dry to obtain 4.6g white solid D-dencichine, yield 73%.
m.p.206℃。
IR(KBr):3503，3436，3386，3241，1671，1634，1517，550cm
-1。
1HNMR(600MHz,D
2O)δ:4.04(1H,dd,J＝7.6Hz,3.6Hz,HOOC-
CH-NH
2),3.87(1H,dd,J＝14.9Hz,3.7Hz,CH-
CH 2 -NH),3.74(1H,dd,J＝14.8Hz,7.7Hz,CH-
CH 2 -NH)；
13CNMR(150MHz,D
2O)δ:177.8,166.1,165.5,54.7,39.9。ESI-HRMS:calcdforC
5H
9N
2O
5,177.1354；found,177.1361.[M+H]。
As an alternative embodiment of the invention, the synthesis of described oxalic acid list potassium ester can comprise the steps: to get dimethyl oxalate (23.6g, 200mmol) in round-bottomed flask, adds 40mL dissolve with methanol, add potassium acetate (19.6g, 200mmol) more fully to stir; Drip (3.6mL subsequently, 200mmol) distilled water, after being warming up to outer bath 80 DEG C of back flow reaction 3h, reaction mixture is cooled to room temperature, then there is a large amount of solid to separate out after adding 40mL ethyl acetate, then carry out suction filtration, dry in cool place place after methyl alcohol, ethyl acetate drip washing, obtain white needle-like crystals oxalic acid list potassium ester 25.9g, yield 91%.
Embodiment 2
1) synthesis of the first intermediate S-D-1:
Get D-Asn one water thing (9.0g, 60mmol) in round-bottomed flask, add distilled water 310mL, add sodium bicarbonate (10.911.2g, 134mmol) subsequently and fully stir; Get 9-fluorene methyl-N-succinimidyl carbonate (21g, 62mmol) be dissolved in 155mL acetone, the acetone soln of described 9-fluorene methyl-N-succinimidyl carbonate is slowly added dropwise in above-mentioned round-bottomed flask, stirring reaction 3.5h at 27 DEG C; Concentrated hydrochloric acid is adopted to regulate the pH value to 2.5 of reaction mixture, then this reaction mixture of suction filtration, obtain 19.8g white solid, be the first intermediate S-D-1, yield 93%.
2) synthesis of the second intermediate S-D-2:
Get (24.9g, 56mmol) [two (trifluoroacetyl oxygen base) iodine] benzene (PIFA) in round-bottomed flask, add 58mL acetonitrile and dissolve; Separately get (19.2g, 54.2mmol) S-D-1, be dissolved in 115mLDMF, and be slowly added dropwise in described round-bottomed flask, vigorous stirring is clarified to reaction solution; Slowly drip subsequently in distilled water 58mL to described round-bottomed flask, after stirring 30min, slowly drip in pyridine 4.6mL to described round-bottomed flask, and at 30 DEG C stirring reaction 48h.Monitor raw material reaction completely by TLC, steam after desolventizing, add distilled water 380mL and stir, the pH value to 2.5 of 6NHCl solution regulator solution; Adopt ether to extract again, water layer 4NNaOH solution adjust pH is that after 2h placed by 7,4 DEG C of refrigerators, suction filtration obtains 12.9g white solid, is the second intermediate S-D-2, yield 73%
3) synthesis of the 3rd intermediate S-D-3:
Get S-D-2 (13.0g, 40mmol) in round-bottomed flask, add dehydrated alcohol 950mL, subsequently to dripping diethylamine 200mL in reaction flask, reacting at 30 DEG C and spending the night.After TLC detection reaction is complete, concentrating under reduced pressure, ethanol band solvent twice.Add anhydrous methanol 100mL stirring and dissolving, drip 5mL methanol hydrochloride solution, after stirring reaction 4h, adularescent solid is separated out, and suction filtration, methanol wash column, obtains 5.1g white solid S-D-3, yield 92%.
4) synthesis of target product D-dencichine:
Get S-D-3 (5.3g, 36mmol) in round-bottomed flask, add 26.5mL50% methanol aqueous solution and stir, in ice bath downhill reaction bottle, add lithium hydroxide one water thing (2.3g, 51mmol) subsequently, stirring reaction 2h; After question response liquid dissolves, slowly drip the 32mL50% methanol aqueous solution of oxalic acid list potassium ester (9.7g, 65mmol), and react 18h under room temperature.After TLC monitoring reacts completely, suction filtration reaction mixture, is dissolved in frozen water by the white solid obtained, and adopts 2NHCl adjust pH to be 2.1 ~ 2.2, make again to separate out white solid, by it after 1.5h placed by 4 DEG C of refrigerators, suction filtration, carries out frozen water successively and washes, methanol wash column, after acetone is washed, dry to obtain 4.8g white solid D-dencichine, yield 75%.
m.p.206℃。
IR(KBr):3503，3436，3386，3241，1671，1634，1517，550cm
-1。
1HNMR(600MHz,D
2O)δ:4.04(1H,dd,J＝7.6Hz,3.6Hz,HOOC-
CH-NH
2),3.87(1H,dd,J＝14.9Hz,3.7Hz,CH-
CH 2 -NH),3.74(1H,dd,J＝14.8Hz,7.7Hz,CH-
CH 2 -NH)；
13CNMR(150MHz,D
2O)δ:177.8,166.1,165.5,54.7,39.9。ESI-HRMS:calcdforC
5H
9N
2O
5,177.1354；found,177.1361.[M+H]。
Embodiment 3
1) synthesis of the first intermediate S-D-1:
Get D-Asn one water thing (9.3g, 62mmol) in round-bottomed flask, add distilled water 320mL, add sodium bicarbonate (11.7g, 139mmol) subsequently and fully stir; Getting 9-fluorene methyl-N-succinimidyl carbonate (21g, 64mmol) is dissolved in 160mL acetone, is slowly added dropwise in above-mentioned round-bottomed flask, stirring reaction 3h at 30 DEG C by the acetone soln of described 9-fluorene methyl-N-succinimidyl carbonate; Concentrated hydrochloric acid is adopted to regulate the pH value to 2.8 of reaction mixture, then this reaction mixture of suction filtration, obtain 20.2g white solid, be the first intermediate S-D-1, yield 92%.
2) synthesis of the second intermediate S-D-2:
Get (25.3g, 56.8mmol) [two (trifluoroacetyl oxygen base) iodine] benzene (PIFA) in round-bottomed flask, add 58mL acetonitrile and dissolve; Separately get (19.5g, 55mmol) S-D-1, be dissolved in 117mLDMF, and be slowly added dropwise in described round-bottomed flask, vigorous stirring is clarified to reaction solution; Slowly drip subsequently in distilled water 60mL to described round-bottomed flask, after stirring 30min, slowly drip in pyridine 4.6mL to described round-bottomed flask, and at 28 DEG C stirring reaction 60h.Monitor raw material reaction completely by TLC, steam after desolventizing, add distilled water 380mL and stir, the pH value to 2.2 of 6NHCl solution regulator solution; Adopt ether to extract again, water layer 4NNaOH solution adjust pH is that after 2.5h placed by 5.2,4 DEG C of refrigerators, suction filtration obtains 13.6g white solid, is the second intermediate S-D-2, yield 76%
3) synthesis of the 3rd intermediate S-D-3:
Get S-D-2 (13.0g, 40mmol) in round-bottomed flask, add dehydrated alcohol 900mL, subsequently to dripping diethylamine 200mL in reaction flask, reacting at 24.5 DEG C and spending the night.After TLC detection reaction is complete, concentrating under reduced pressure, ethanol band solvent twice.Add anhydrous methanol 100mL stirring and dissolving, drip 5mL methanol hydrochloride solution, after stirring reaction 4.5h, adularescent solid is separated out, and suction filtration, methanol wash column, obtains 5.0g white solid S-D-3, yield 91%.
4) synthesis of target product D-dencichine:
Get S-D-3 (5.3g, 36mmol) in round-bottomed flask, add 26.5mL50% methanol aqueous solution and stir, in-5 DEG C of downhill reaction bottles, add lithium hydroxide one water thing (2.3g, 51mmol) subsequently, stirring reaction 2h; After question response liquid dissolves, slowly drip the 32mL50% methanol aqueous solution of oxalic acid list potassium ester (9.7g, 65mmol), and react 15h under room temperature.After TLC monitoring reacts completely, suction filtration reaction mixture, is dissolved in frozen water by the white solid obtained, and adopts 2NHCl adjust pH to be 2.1 ~ 2.2, make again to separate out white solid, by it after 2h placed by 4 DEG C of refrigerators, suction filtration, carries out frozen water successively and washes, methanol wash column, after acetone is washed, dry to obtain 4.7g white solid D-dencichine, yield 74%.
m.p.206℃。
IR(KBr):3503，3436，3386，3241，1671，1634，1517，550cm
-1。
1HNMR(600MHz,D
2O)δ:4.04(1H,dd,J＝7.6Hz,3.6Hz,HOOC-
CH-NH
2),3.87(1H,dd,J＝14.9Hz,3.7Hz,CH-
CH 2 -NH),3.74(1H,dd,J＝14.8Hz,7.7Hz,CH-
CH 2 -NH)；
13CNMR(150MHz,D
2O)δ:177.8,166.1,165.5,54.7,39.9。ESI-HRMS:calcdforC
5H
9N
2O
5,177.1354；found,177.1361.[M+H]。
Embodiment 4
1) synthesis of the first intermediate S-D-1:
Get D-Asn one water thing (9.6g, 64mmol) in round-bottomed flask, add distilled water 330mL, add sodium bicarbonate (10.9g, 143mmol) subsequently and fully stir; Getting 9-fluorene methyl-N-succinimidyl carbonate (22g, 66mmol) is dissolved in 165mL acetone, is slowly added dropwise in above-mentioned round-bottomed flask, stirring reaction 8h at 20 DEG C by the acetone soln of described 9-fluorene methyl-N-succinimidyl carbonate; Concentrated hydrochloric acid is adopted to regulate the pH value to 3.0 of reaction mixture, then this reaction mixture of suction filtration, obtain 20.4g white solid, be the first intermediate S-D-1, yield 90%.
2) synthesis of the second intermediate S-D-2:
Get (24.9g, 56mmol) [two (trifluoroacetyl oxygen base) iodine] benzene (PIFA) in round-bottomed flask, add 58mL acetonitrile and dissolve; Separately get (19.2g, 54.2mmol) S-D-1, be dissolved in 115mLDMF, and be slowly added dropwise in described round-bottomed flask, vigorous stirring is clarified to reaction solution; Slowly drip subsequently in distilled water 58mL to described round-bottomed flask, after stirring 20min, slowly drip in pyridine 4.6mL to described round-bottomed flask, and at 40 DEG C stirring reaction 32h.Monitor raw material reaction completely by TLC, steam after desolventizing, add distilled water 380mL and stir, the pH value to 2 of 6NHCl solution regulator solution; Adopt ether to extract again, water layer 4NNaOH solution adjust pH is that after 3h placed by 6.5,4 DEG C of refrigerators, suction filtration obtains 12.4g white solid, is the second intermediate S-D-2, yield 70%
3) synthesis of the 3rd intermediate S-D-3:
Get S-D-2 (13.7g, 42mmol) in round-bottomed flask, add dehydrated alcohol 1000mL, subsequently to dripping diethylamine 200mL in reaction flask, reacting at 27 DEG C and spending the night.After TLC detection reaction is complete, concentrating under reduced pressure, ethanol band solvent twice.Add anhydrous methanol 100mL stirring and dissolving, drip 5mL methanol hydrochloride solution, after stirring reaction 4h, adularescent solid is separated out, and suction filtration, methanol wash column, obtains 5.2g white solid S-D-3, yield 93%.
4) synthesis of target product D-dencichine:
Get S-D-3 (4.7g, 32mmol) in round-bottomed flask, add 25mL50% methanol aqueous solution and stir, in ice bath downhill reaction bottle, add lithium hydroxide one water thing (2.0g, 45mmol) subsequently, stirring reaction 1h; After question response liquid dissolves, slowly drip the 30mL50% methanol aqueous solution of oxalic acid list potassium ester (8.7g, 58mmol), and react 18h under room temperature.After TLC monitoring reacts completely, suction filtration reaction mixture, is dissolved in frozen water by the white solid obtained, and adopts 2NHCl adjust pH to be 2.1 ~ 2.2, make again to separate out white solid, by it after 2h placed by 4 DEG C of refrigerators, suction filtration, carries out frozen water successively and washes, methanol wash column, after acetone is washed, dry to obtain 4.9g white solid D-dencichine, yield 77%.
m.p.206℃。
IR(KBr):3503，3436，3386，3241，1671，1634，1517，550cm
-1。
1HNMR(600MHz,D
2O)δ:4.04(1H,dd,J＝7.6Hz,3.6Hz,HOOC-
CH-NH
2),3.87(1H,dd,J＝14.9Hz,3.7Hz,CH-
CH 2 -NH),3.74(1H,dd,J＝14.8Hz,7.7Hz,CH-
CH 2 -NH)；
13CNMR(150MHz,D
2O)δ:177.8,166.1,165.5,54.7,39.9。ESI-HRMS:calcdforC
5H
9N
2O
5,177.1354；found,177.1361.[M+H]。
Embodiment 5
1) synthesis of the first intermediate S-D-1:
Get D-Asn one water thing (8.7g, 66mmol) in round-bottomed flask, add distilled water 300mL, add sodium bicarbonate (10.9g, 130mmol) subsequently and fully stir; Get 9-fluorene methyl-N-succinimidyl carbonate (20g, 60mmol) be dissolved in 150mL acetone, the acetone soln of described 9-fluorene methyl-N-succinimidyl carbonate is slowly added dropwise in above-mentioned round-bottomed flask, stirring reaction 3.5h at 27 DEG C; Concentrated hydrochloric acid is adopted to regulate the pH value to 2.5 of reaction mixture, then this reaction mixture of suction filtration, obtain 20.2g white solid, be the first intermediate S-D-1, yield 95%.
2) synthesis of the second intermediate S-D-2:
Get (24.9g, 56mmol) [two (trifluoroacetyl oxygen base) iodine] benzene (PIFA) in round-bottomed flask, add 58mL acetonitrile and dissolve; Separately get (19.2g, 54.2mmol) S-D-1, be dissolved in 115mLDMF, and be slowly added dropwise in described round-bottomed flask, vigorous stirring is clarified to reaction solution; Slowly drip subsequently in distilled water 58mL to described round-bottomed flask, after stirring 20min, slowly drip in pyridine 4.6mL to described round-bottomed flask, and at 24 DEG C stirring reaction 70h.Monitor raw material reaction completely by TLC, steam after desolventizing, add distilled water 380mL and stir, the pH value to 2.5 of 6NHCl solution regulator solution; Adopt ether to extract again, water layer 4NNaOH solution adjust pH is that after 2h placed by 7,4 DEG C of refrigerators, suction filtration obtains 13.3g white solid, is the second intermediate S-D-2, yield 75%
3) synthesis of the 3rd intermediate S-D-3:
Get S-D-2 (13.0g, 40mmol) in round-bottomed flask, add dehydrated alcohol 1000mL, subsequently to dripping piperidinyl-1 50mL in reaction flask, reacting at 27 DEG C and spending the night.After TLC detection reaction is complete, concentrating under reduced pressure, ethanol band solvent twice.Add anhydrous methanol 100mL stirring and dissolving, drip 5mL methanol hydrochloride solution, after stirring reaction 4h, adularescent solid is separated out, and suction filtration, methanol wash column, obtains 4.7g white solid S-D-3, yield 85%.
4) synthesis of target product D-dencichine:
Get S-D-3 (5.3g, 36mmol) in round-bottomed flask, add 26.5mL50% methanol aqueous solution and stir, in ice bath downhill reaction bottle, add lithium hydroxide one water thing (2.3g, 51mmol) subsequently, stirring reaction 1.2h; After question response liquid dissolves, slowly drip the 32mL50% methanol aqueous solution of oxalic acid list potassium ester (9.7g, 65mmol), and react 24h under room temperature.After TLC monitoring reacts completely, suction filtration reaction mixture, is dissolved in frozen water by the white solid obtained, and adopts 2NHCl adjust pH to be 2.1 ~ 2.2, make again to separate out white solid, by it after 2h placed by 4 DEG C of refrigerators, suction filtration, carries out frozen water successively and washes, methanol wash column, after acetone is washed, dry to obtain 4.6g white solid D-dencichine, yield 73%.
m.p.206℃。
IR(KBr):3503，3436，3386，3241，1671，1634，1517，550cm
-1。
1HNMR(600MHz,D
2O)δ:4.04(1H,dd,J＝7.6Hz,3.6Hz,HOOC-
CH-NH
2),3.87(1H,dd,J＝14.9Hz,3.7Hz,CH-
CH 2 -NH),3.74(1H,dd,J＝14.8Hz,7.7Hz,CH-
CH 2 -NH)；
13CNMR(150MHz,D
2O)δ:177.8,166.1,165.5,54.7,39.9。ESI-HRMS:calcdforC
5H
9N
2O
5,177.1354；found,177.1361.[M+H]。
Embodiment 6
1) synthesis of the first intermediate S-D-1:
Get D-Asn one water thing (8.7g, 66mmol) in round-bottomed flask, add distilled water 300mL, add sodium bicarbonate (10.9g, 130mmol) subsequently and fully stir; Get 9-fluorene methyl-N-succinimidyl carbonate (20g, 60mmol) be dissolved in 150mL acetone, the acetone soln of described 9-fluorene methyl-N-succinimidyl carbonate is slowly added dropwise in above-mentioned round-bottomed flask, stirring reaction 3.5h at 27 DEG C; Concentrated hydrochloric acid is adopted to regulate the pH value to 2.5 of reaction mixture, then this reaction mixture of suction filtration, obtain 20.2g white solid, be the first intermediate S-D-1, yield 95%.
2) synthesis of the second intermediate S-D-2:
Get (24.9g, 56mmol) [two (trifluoroacetyl oxygen base) iodine] benzene (PIFA) in round-bottomed flask, add 58mL acetonitrile and dissolve; Separately get (19.2g, 54.2mmol) S-D-1, be dissolved in 115mLDMF, and be slowly added dropwise in described round-bottomed flask, vigorous stirring is clarified to reaction solution; Slowly drip subsequently in distilled water 58mL to described round-bottomed flask, after stirring 20min, slowly drip in pyridine 4.6mL to described round-bottomed flask, and at 24 DEG C stirring reaction 70h.Monitor raw material reaction completely by TLC, steam after desolventizing, add distilled water 380mL and stir, the pH value to 2.5 of 6NHCl solution regulator solution; Adopt ether to extract again, water layer 4NNaOH solution adjust pH is that after 2h placed by 7,4 DEG C of refrigerators, suction filtration obtains 13.3g white solid, is the second intermediate S-D-2, yield 75%
3) synthesis of the 3rd intermediate S-D-3:
Get S-D-2 (13.0g, 40mmol) in round-bottomed flask, add dehydrated alcohol 1000mL, subsequently to dripping piperidinyl-1 30mL in reaction flask, reacting at 27 DEG C and spending the night.After TLC detection reaction is complete, concentrating under reduced pressure, ethanol band solvent twice.Add anhydrous methanol 100mL stirring and dissolving, drip 5mL methanol hydrochloride solution, after stirring reaction 4h, adularescent solid is separated out, and suction filtration, methanol wash column, obtains 4.6g white solid S-D-3, yield 83%.
4) synthesis of target product D-dencichine:
Get S-D-3 (5.3g, 36mmol) in round-bottomed flask, add 26.5mL50% methanol aqueous solution and stir, in ice bath downhill reaction bottle, add lithium hydroxide one water thing (2.3g, 51mmol) subsequently, stirring reaction 1.2h; After question response liquid dissolves, slowly drip the 32mL50% methanol aqueous solution of oxalic acid list potassium ester (9.7g, 65mmol), and react 24h under room temperature.After TLC monitoring reacts completely, suction filtration reaction mixture, is dissolved in frozen water by the white solid obtained, and adopts 2NHCl adjust pH to be 2.1 ~ 2.2, make again to separate out white solid, by it after 2h placed by 4 DEG C of refrigerators, suction filtration, carries out frozen water successively and washes, methanol wash column, after acetone is washed, dry to obtain 4.6g white solid D-dencichine, yield 73%.
m.p.206℃。
IR(KBr):3503，3436，3386，3241，1671，1634，1517，550cm
-1。
1HNMR(600MHz,D
2O)δ:4.04(1H,dd,J＝7.6Hz,3.6Hz,HOOC-
CH-NH
2),3.87(1H,dd,J＝14.9Hz,3.7Hz,CH-
CH 2 -NH),3.74(1H,dd,J＝14.8Hz,7.7Hz,CH-
CH 2 -NH)；
13CNMR(150MHz,D
2O)δ:177.8,166.1,165.5,54.7,39.9。ESI-HRMS:calcdforC
5H
9N
2O
5,177.1354；found,177.1361.[M+H]。
The short thrombocyte of D-dencichine increases biological activity test: following content is measure the impact by above-described embodiment gained D-dencichine, carboplatin being caused to mouse and rat platelet minimizing model.
1.D-dencichine causes on carboplatin the impact that mouse platelets reduces model
Carboplatin is the clinical medicine being used for the treatment of tumour, and it shows obvious bone marrow depression antineoplastic simultaneously, and severity has obvious dose-effect relationship, wherein particularly outstanding to megalokaryocyte-platelet system restraining effect.This test adopts carboplatin to cause thrombopenia mouse model, injection regrouping human interleukin-11 (IL-11) is positive control, observe the impact of D-dencichine human peripheral blood platelet number and hemopoietic function of bone marrow, specify it and thrombocytopenic action effect is caused to chemotherapeutic.
Experimental technique is as follows:
Get 48 mouse, after conforming, eye socket gets blood, detects intact animal routine blood test.Six groups are divided at random: blank group, model group, interleukin-11 (0.37mg/kg) dosage group, L-dencichine group (3.0mg/kg), D-dencichine (3.0,0.2mg/kg) two groups of dosed administration groups by hemocyte and body weight homeostatic principle.Then by above-mentioned dosage and route of administration administration respectively, once a day, successive administration 14 days, blank group and model group give same volume physiological saline 10mL/kg simultaneously.Administration got blood after 14 days, detected routine blood test.All the other each treated animals equal intravenous injection carboplatin 120mg/kg modeling except blank group in second day, blank group intravenous injection simultaneously same volume physiological saline 8mL/kg; Give trial drug 7 days continuously with aforesaid method again, and before each blood sampling, animal is weighed, within the 7th, 10 days, get blood respectively at injection carboplatin, detect routine blood test.Within 7th day, give investigational agent after injection carboplatin after, namely stop continuing administration.Injection carboplatin the 10th day animal is weighed, and dislocation is put to death, and gets right side femur, does marrow protection inspection.
Experimental result is as follows:
Impact on thrombocyte (PLT): from table 1 result, in advance administration after 14 days D-dencichine 3.0mg/kg dosage group PLT than obviously increase with blank group, interleukin-11 group, L-dencichine 3.0mg/kg and D-dencichine 0.2mg/kg dosed administration treated animal PLT and blank group compare the trend all having increase; After the 7th day, model group PLT is significantly lower than blank group for injection carboplatin, and interleukin-11 and each group of D-dencichine are apparently higher than model group and L-dencichine group; Injection carboplatin the 10th day, model group PLT is still starkly lower than blank group, reduces by 92%, and D-dencichine 3.0mg/kg dosage group PLT comparatively model group obviously increase, have significant difference.
Table 1. dencichine causes thrombopenia model mice thrombocyte (× 10 to carboplatin
9individual/L) impact
Note: compare with blank group, * p<0.05, * * p<0.01, * * * p<0.001; Compare with model group, #p<0.05, ##p<0.01, ###p<0.001.
2.D-dencichine causes on carboplatin the impact that rat platelet reduces model
Above-mentioned mouse test confirms that D-dencichine drug administration by injection can significantly improve peripheral blood PLT and count.This test adopts carboplatin intravenous injection to prepare P of Rats LT minimizing model, by observing the change of the hematological indices of rat, platelet function, myeloid element function, Morphology of Bone Marrow pathology and Some cytokines, confirm that D-dencichine reduces the curative effect of disease and preliminary mechanism of action to P of Rats LT further.
Experimental technique is as follows:
Get 36 rats, after conforming, eye socket gets blood, detects intact animal routine blood test.Six groups are divided at random: blank group, model group, interleukin-11 (0.26 μ g/kg) dosage group, L-dencichine group (2.1mg/kg), D-dencichine (2.1,0.26mg/kg) two groups of dosed administration groups by hemocyte and body weight homeostatic principle.Then be administered once every day respectively by above-mentioned dosage and route of administration, successive administration 7 days, blank group and model group give same volume physiological saline 10mL/kg simultaneously.Thereafter all the other each treated animals equal intravenous injection injection carboplatin 40mg/kg modeling except blank group, blank group intravenous injection simultaneously same volume physiological saline 6mL/kg; Give trial drug 7 days continuously with aforesaid method again, and before each blood sampling, animal is weighed.Within 7th day, give investigational agent after injection carboplatin after, namely stop continuing administration.Within 7th, 10 days, get blood respectively at injection carboplatin, detect routine blood test.Injection carboplatin the 10th day animal is weighed pneumoretroperitoneum injection Chloral Hydrate 300mg/kg anesthesia, gets blood, marrow does coherence check.
Experimental result is as follows:
Impact on thrombocyte (PLT): from table 2 result, in advance administration is PLT interleukin-11 group and L-dencichine and D-dencichine 2.1mg/kg group and the obvious rising of blank group after 7 days; Injection carboplatin the 7th day, PLT model group compares with blank group to reduce and obviously reduces, and D-dencichine two dosage group 2.1mg/kg, 0.26mg/kg, PLT are all apparently higher than model group; Injection carboplatin the 10th day, PLT model group and the obvious reduction of blank group, be equivalent to 3% of blank group, same D-dencichine two dosage group 2.1mg/kg, 0.26mg/kg are all apparently higher than model group, wherein under same concentration (0.26mg/kg), the rising trend of D-dencichine group to PLT is obviously better than positive control drug interleukin-11 group, demonstrates and well suppresses thrombopenia active.
Table 2. dencichine causes thrombopenia rat model thrombocyte (× 10 to carboplatin
9individual/L) impact
Note: compare with blank group, * p<0.05, * * p<0.01, * * * p<0.001; Compare with model group, #p<0.05, ##p<0.01, ###p<0.001.
As can be seen here, the present invention adopts the preparation method of highly effective and safe to synthesize and obtains D-dencichine, preparation technology is simple, yield is higher, Compound D-the dencichine prepared by method provided by the invention has treatment thrombocytopenia effect preferably, be better than clinical application interleukin-11, the drug candidate for the treatment of thrombocytopenia can be become.
Those of ordinary skill in the field are to be understood that: the foregoing is only specific embodiments of the invention; be not limited to the present invention; within the spirit and principles in the present invention all, any amendment made, equivalent replacement, improvement etc., all should be included within protection scope of the present invention.
Claims (10)
1. a preparation method for D-dencichine, is characterized in that, comprises the following steps:
1) Fmoc protection is carried out to the amino of D-Asn, obtain the first intermediate;
2) Hoffman DeR is carried out to described first intermediate, obtain the second intermediate;
3) under the effect of organic bases, described second intermediate is sloughed Fmoc protection, obtain the 3rd intermediate;
4) under basic conditions, described 3rd intermediate and oxalic acid list potassium ester are carried out condensation reaction, obtains described D-dencichine.
2. the preparation method of D-dencichine according to claim 1, is characterized in that, adopts 9-fluorene methyl-N-succinimidyl carbonate to carry out Fmoc protection to the amino of D-Asn.
3. the preparation method of D-dencichine according to claim 1, is characterized in that, adopts [two (trifluoroacetyl oxygen base) iodine] benzene to carry out Hoffman DeR to described first intermediate, obtains the second intermediate.
4. the preparation method of D-dencichine according to claim 1, is characterized in that, described organic bases is selected from diethylamine or piperidines.
5. the preparation method of D-dencichine according to claim 1, is characterized in that, described highly basic is selected from lithium hydroxide.
6. the preparation method of D-dencichine according to claim 1, is characterized in that, the preparation method of described first intermediate comprises the following steps:
Sodium bicarbonate is joined in D-Asn, 9-fluorene methyl-N-succinimidyl carbonate is dissolved in acetone;
The acetone soln of described 9-fluorene methyl-N-succinimidyl carbonate is slowly added dropwise in the mixing solutions of sodium bicarbonate, D-Asn, reacts at 20-30 DEG C;
Adopt acid solution to regulate the pH value of reaction mixture to 2-4, then this reaction mixture of suction filtration, obtains white solid, is described first intermediate.
7. the preparation method of D-dencichine according to claim 1, is characterized in that, the preparation method of described second intermediate comprises the following steps:
[two (trifluoroacetyl oxygen base) iodine] benzene is dissolved in acetonitrile, described first intermediate is dissolved in DMF, the described DMF being dissolved with the first intermediate is joined in acetonitrile solution;
Continue to add water to acetonitrile solution, and then continue to add pyridine to acetonitrile solution, react at 20-30 DEG C;
After reaction terminates, steaming desolventizes, and then adds water, adopts acid solution to regulate the pH value of reaction mixture to 1-3;
Then, adopt ether to extract, water layer adopts lye pH adjustment value to 5-7, and after refrigeration, suction filtration obtains white solid, is described second intermediate.
8. the preparation method of D-dencichine according to claim 1, is characterized in that, the preparation method of described 3rd intermediate comprises the following steps:
Described second intermediate is dissolved in ethanol, then continues to add organic bases, react at 20-30 DEG C;
After reaction terminates, concentrating under reduced pressure reaction mixture, adopts washing with alcohol;
Add methyl alcohol and methanol hydrochloride solution to the reaction mixture after washing with alcohol, after adularescent solid is separated out, obtain white solid through suction filtration, methanol wash, be described 3rd intermediate.
9. a pharmaceutical composition, is characterized in that, comprises the D-dencichine obtained according to the preparation method of the D-dencichine in claim 1 ~ 8 described in any one.
10. the purposes of a pharmaceutical composition according to claim 9 in preparation treatment thrombopenia disease drug.
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| CN109180532A (en) * | 2018-08-29 | 2019-01-11 | 南京天际联盟医药科技有限公司 | The high efficiency preparation method of D- dencichine |
| CN112125819A (en) * | 2020-08-19 | 2020-12-25 | 兰州百源基因技术有限公司 | Preparation method of sanchinin |
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