CN109134555A - Anticoagulant five saccharide compound and preparation method thereof and medical usage - Google Patents
Anticoagulant five saccharide compound and preparation method thereof and medical usage Download PDFInfo
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- C07H15/00—Compounds containing hydrocarbon or substituted hydrocarbon radicals directly attached to hetero atoms of saccharide radicals
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- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
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Abstract
The present invention relates to anticoagulation medicines, and in particular to one kind five sugar compounds of synthesis and its salt, preparation method and its pharmaceutical applications.The present invention has synthesized five sugar compounds or its sour form or its salt form of a kind of ionic species as shown in formula III, with compared with the stronger anticoagulin xa activity of Fondaparinux sodium and longer elimination half-life period.Five sugar compounds of the invention can be used for preparing the purposes in the drug of prevention and treatment and blood coagulation disorder related disease, such as diseases such as Deep vain thrombosis, thrombophlebitis, the artery occlusion as caused by thrombosis or embolism, Postoperative Intravenous thrombosis or embolisms.
Description
Technical field
The present invention relates to five saccharide compounds for being used as anticoagulant, preparation method and its doctor in terms of anticoagulation
Medicinal way.
Background technique
Thrombotic diseases are a kind of diseases for seriously endangering human health, and disease incidence is in first of various diseases, in recent years
Carry out also cumulative gesture.It is broadly divided into arterial thrombus and phlebothrombosis.Phlebothrombosis is common in Deep venou, and clinical manifestation is blood
Caused embolism causes organ function to hinder after local pain swelling, distal end blood backflow obstacle and thrombus breaks loose that bolt is formed
Hinder.Arterial thrombosis is can lead to serious since arterial blood tube wall atherosclerotic lesion and platelet activation
Cardiovascular disease such as acute myocardial infarction AMI, cerebral apoplexy etc..Treatment method includes that anticoagulant therapy, Antiplatelet therapy and thrombolysis are controlled
Treat etc., and anticoagulant therapy is core and the basis of current clinical prevention and treatment thrombotic diseases.
Heparin is traditional anticoagulant, is one of glycosaminoglycan family polysaccharide.What is clinically used includes common
Heparin and low molecular weight heparin.The conformation that heparin induces AT and being attached to the particular combination domain of antithrombase (AT) becomes
Change, and then inhibits the activity of factor Xa.Studies have shown that the Heparin-binding AT and minimum structural unit for inhibiting Xa factor is one
A unique five glycosylation sequence.
Up to the present, have more documents and disclose and synthesize to obtain with antithrombus formation and anticoagulation by full chemistry
Active five sugar compounds.Such as United States Patent (USP) US4818816, Carbohydrate Research 1987 (167): 67-75 report
The Fondaparinux sodium in road is the first generation synthetic analogues that five glycosylation sequences are combined based on anticoagulant heparin hemase, is belonged to indirectly solidifying
Blood factor Xa inhibitor.Fondaparinux sodium is about 0.7 hour in rat intracorporal half-life period, intracorporal half-life period is about 17 in people
Hour.The product since U.S.'s listing, in multiple country's listings, were clinically used to treat and prevent Deep vein from 2002
Thromboembolic events occur, recommended dose once a day, subcutaneous administrations.Although Fondaparinux sodium clinical manifestation is outstanding,
It is that its synthesis difficulty is very big, production cost is high.
United States Patent (USP) US5378829, which is reported, replaces N- in Fondaparinux sodium structure with alkoxy grp and O- sulfonate group
Sulfonate group, hydroxyl group and a new class of pentasaccharides formed, such as idraparinux sodium (Idraparinux).Due to pentasaccharides list
Alkoxy base is introduced in member and O- sulfonate group, preparation mode are greatly simplified.It is 9.2 in rat Half-life in vivo
Hour (iv) dramatically increases bleeding risk in the intracorporal Increased Plasma Half-life of people to 120h, thus stops at phase III clinical trial.
WO9936428 is reported by replacing O- alkyl with alkylidene bridge and locking the configuration of G unit L- iduronic acid
And five sugar compounds of one kind prepared, these compounds have strong affine with effective anticoagulin xa activity and to AT
Power.
WO9925720 discloses a kind of five sugar compounds with 4~6 sulfate radicals.The reduction of sulfate groups is not
Antithrombotic activity is reduced, the decrease of platelet that heparin will not be caused to induce.
WO0224754 discloses five sugar compounds of one kind with biotin or biotin derivative covalent linkage.Biotin
Or derivatives thereof introducing there is no change pentasaccharides pharmaceutical activity, it is advantageous that such compound also has in emergency situations
The advantages of inhibiting its anticoagulant active, is neutralized by avidin solution agent rapidly.
WO2012172104 discloses five sugar compounds of high activity of a kind of half-life short.By the substitution for changing D unit
Base regulates and controls alkylation/O- sulphation pentasaccharides half-life period.
The half-life period of anticoagulation medicine is very important pharmacokinetic parameter.In clinic in the case where bleeding occurs
Need to close blood coagulation resisting function as quickly as possible, and the half-life period too long bleeding risk that will lead to increases.And for certain thrombus bolts
Plug property disease, such as pulmonary embolism, venae profunda conducted, non-ST elevation acute myocardial infraction, suitably extend the half-life period of anticoagulant
It is more favorable.
On the other hand, the core of anticoagulation medicine research and development is safety issue, how effectively to avoid hemorrhagic tendency, reduces out
Blood risk is the exploitation institute facing challenges of anticoagulation medicine.
Summary of the invention
In order to solve the problems in the existing technology, the present invention provides it is a kind of complete it is newly synthesized as shown in formula III from
Five sugar compounds of sub- form or its acid or its salt:
Wherein, work as R1When for H, R2For H or SO3 -；
Work as R1For SO3 -Or when C1~C4 alkyl, R2For H.
Preferably, five sugar compounds and its salt, anionic form are selected from such as flowering structure:
Wherein R1For C1~C4 alkyl, preferably methyl.
The present invention includes the synthesis pentasaccharides and its salt form of sour the form ,-COO in sour form-With-OSO3 -Functional group's difference
It is-COOH form and-OSO3H-shaped formula；Salt form of the invention is selected from sodium salt, sylvite, calcium salt or ammonium salt, particular certain cancers or potassium
Salt.
Further, the present invention provides the preparation methods of five sugar compounds, comprising the following steps:
(a) disaccharides shown in formula DE is reacted into preparation formula DEFGH0 with trisaccharide shown in formula FGH,
Wherein, R1Selected from Ac, Bn, C1~C4 alkyl, R2Selected from Bn, Bz.
(b) formula DEFGH0 is removed into benzyl with preparation formula DEFGH1,
Wherein, RaFor Ac, H, C1~C4 alkyl, RbFor H, Bz；
(c) sulfating reaction preparation formula DEFGH2 is occurred into for formula DEFGH1 depicted compound,
Wherein, wherein RcFor Ac, SO3 -, C1~C4 alkyl, RdFor SO3 -, Bz；
(d) by compound hydrolysis preparation formula III-1~III-4 shown in formula DEFGH2.
The present invention also provides preparation five sugar compounds and its salt intermediate DE,
Wherein, R1Selected from Ac, Bn, C1~C4 alkyl.
And following formula intermediate,
Wherein, R2Selected from Bn, Bz.
And following formula intermediate,
Wherein, R1Selected from Ac, Bn, C1~C4 alkyl, R2Selected from Bn, Bz.
And following formula intermediate,
Wherein, RaFor Ac, H, C1~C4 alkyl, RbFor H, Bz.
The present invention also provides following formula intermediate,
Wherein, RcFor Ac, SO3 -, C1~C4 alkyl, RdFor SO3 -, Bz.
On the other hand, the invention further relates to pharmaceutical composition, including containing the compounds of this invention as active constituent with
And optional pharmaceutically acceptable auxiliary material.
In each unit dose, five sugar compounds shown in formula III of the invention, its acid or its salt be 0.1~
100mg, preferably 0.5~50mg.
Described pharmaceutical composition can be administered by oral or extra-parenteral approach, and parenteral route includes but unlimited
In intravenous injection, intramuscular injection and subcutaneous injection.
For oral administration, pharmaceutical composition of the invention provides usually in the form of tablet, capsule, solution.
Tablet may include five sugar compounds shown in formula III of the invention, its acid or its salt and pharmaceutically acceptable excipient.
The excipient includes but is not limited to diluent, disintegrating agent, adhesive, lubricant, sweetener, corrigent, colorant or anti-corrosion
Agent.Diluent can be selected from starch, lactose；Disintegrating agent can be selected from cornstarch, alginic acid；Adhesive can be selected from starch, povidone,
Gelatin；Lubricant can be selected from magnesium stearate, talcum powder, sodium stearyl fumarate.Capsule includes hard capsule and soft capsule.
In hard capsule, five sugar compounds shown in the formula III of the invention as active constituent, its acid or its salt and diluent are mixed
It closes, in soft capsule, five sugar compounds shown in formula III of the invention, its acid or its salt and water or oil such as peanut oil or olive
The mixing of olive oil.
For parenteral route, pharmaceutical composition of the invention can by intravenous injection, intramuscular injection or
Subcutaneous administrations.It is usually provided with aseptic aqueous solution or suspension or freeze-dried powder, and adjusts suitable pH and isotonicity.
In addition, can optional auxiliary material such as mannitol, cellulose derivative, lecithin.Preferred administration route is subcutaneous administrations.
On the other hand, the invention further relates to five sugar compounds shown in formula III, its acid or its salt to prepare for preventing
And/or the purposes in the drug of relevant to the disturbances of blood coagulation disease for the treatment of or morbid state.
In another aspect, the invention further relates to for preventing and/or treating disease relevant to blood coagulation disorder and disease
The method of state comprising given to individual in need five sugar compounds shown in formula III of the invention, its acid or its salt or
Pharmaceutical composition of the invention.
Also on the one hand, the invention further relates to for preventing and/or treating disease relevant to blood coagulation disorder and disease
Five sugar compounds shown in the formula III of the invention of state, its acid or its salt or pharmaceutical composition of the invention.
The disease relevant to coagulation function obstacle or the example of morbid state include but is not limited to venous thronbosis,
Especially Deep vain thrombosis, Arterial thrombosis, thrombophlebitis, pulmonary embolism, acute coronary syndrome, cardiac muscle
Infraction or apoplexy, and the artery occlusion as caused by thrombosis or embolism, angioplasty or thrombolysis process in or it
Artery afterwards blocks again, Postoperative Intravenous thrombosis or embolism, acute or chronic artery sclerosis, ST sections, Non-ST Elevation Acute cardiac muscle
Infarct.In haemodialysis and surgical operation, the compound of the present invention can also be used in the anticoagulant in extracorporeal circulation of blood.Also
The compound of the present invention can be used for the prevention of the relevant ischemic damage and reperfusion damage of solid organ transplantation.
Advantages of the present invention is mainly reflected in: the present invention has synthesized a series of novel by change E ring, the substituent group of H ring
Five sugar compounds, it is amazing be five sugar compounds of the invention have unexpected technical effect.Firstly, of the invention
Five sugar compounds have compared with the stronger anticoagulin xa activity of Fondaparinux sodium, dosage is smaller, so that side effect is more
It is small；Secondly, five sugar compounds of the invention are extended compared with the Half-life in vivo of Fondaparinux sodium, can achieve every 2~3 days to
Medicine is primary, is especially suitable for the treatment of pulmonary embolism, venae profunda conducted, non-ST elevation acute myocardial infraction patient；In preliminary safety
In assessment, the compounds of this invention does not almost make significant difference to hemorrhagic tendency, shows important potential using value；Furthermore this
The preparation method of invention compound greatly simplifies, and manufacturing cost is low, significantly reduces the development cost of bulk pharmaceutical chemicals, is very suitable to work
Industry mass production.
Specific embodiment
Present inventor has synthesized five saccharide compound shown in general formula III, and given birth to by further investigation
Object experimental study finds that five saccharide compounds of the invention have high anticoagulin xa activity and suitably disappear in vivo
Except half-life period, be especially suitable for anticoagulant.
Definition
Unless stated to the contrary, following that there are following meanings with term in the specification and in the claims
" alkyl " refers to the aliphatic hydrocarbon group of saturation, the saturation monovalent hydrocarbon including linear chain or branched chain, and the alkyl has institute
The carbon atom number shown.If term " C1~C4 alkyl " includes C1 alkyl, C2 alkyl, C3 alkyl, C4 alkyl, suitable alkyl base
Group includes methyl, ethyl, n-propyl, isopropyl, normal-butyl, isobutyl group, tert-butyl.
Claims and abbreviation used in the description its meaning see the table below:
| Bz | Benzoyl |
| Bn | Benzyl |
| Ac | Acetyl group |
| DMAP | 4-dimethylaminopyridine |
| DBU | 11 carbon -7- alkene of 1,8- diazabicylo |
| TEMPO | 2,2,6,6- tetramethyl piperidine-nitrogen-oxide |
| BAIB | Iodobenzene diethylester |
| TCCA | Sym-closene |
| TMSOTf | Trimethylsilyl trifluoromethanesulfonate |
| Dowex | Ion exchange resin |
| Sephadex | Sephadex |
Preparation method:
The preparation method of five sugar compounds of the invention is described more particularly below, but these specific preparation methods are not right
The scope of the present invention constitutes any restrictions.In addition, reaction condition such as reactant, solvent, alkali, the amount of compound used therefor, reaction temperature
Degree, reaction time etc. are not limited to following example.
Five sugar compounds of the invention can also optionally will be describing or known in the art various in the present specification
Synthetic method combines and is conveniently made, and such combination can be easy to carry out by those skilled in the art.
The synthesis of first part's monosaccharide D ring
Synthetic route:
A) dimethyl suflfate, KOH, acetonitrile, b) acetic acid, sulfuric acid, water, c) Tritox, DBU, methylene chloride
The preparation of compound D1: tetrahydrofuran dissolution is added in D0 (748g), excessive potassium hydroxide does alkali, is added dropwise at 0 DEG C
Dimethyl suflfate reacts 2h, has reacted plus 4h is quenched in potassium hydroxide aqueous solution, and then ethyl acetate extracts liquid separation, is spin-dried for obtaining D1
(804g, yield 100%).
The preparation of compound D2: acetic acid: water is added in D1 (804g): in sulfuric acid=2000g:400g:98g, back flow reaction
Add ethyl acetate and water extraction after 1h, organic phase is evaporated to obtain D2 (690g, yield 89%).
The preparation of compound D3: D2 (388g) adds anhydrous methylene chloride to dissolve, and it is anti-that Tritox 288g, DBU 15g is added
1h is answered, dry chromatography is then revolved and obtains D3 (489g, yield 92%).
The preparation of second part monosaccharide E ring
Synthetic route:
A) dimethyl suflfate, KOH, tetrahydrofuran, b) trifluoracetic acid, water, c) para-methylbenzenepyridinsulfonate sulfonate, benzaldehyde diformazan
Acetal, d) ethyl acetate, aceticanhydride, triethylamine, DMAP, e) boron trifluoride ether, p methoxy phenol, f) potassium hydroxide, g) system
Standby E8: potassium hydroxide, dimethyl suflfate, h) acetic acid, water, i) TEMPO, BAIB, methylene chloride, water, j) iodomethane, bicarbonate
Potassium, acetonitrile
The preparation of compound E1-E4: Tetrahedron, 2012,68 (36), 7386-7399 are referred to.
The preparation of compound E5:
E4 (366g) and p methoxy phenol 130g are dissolved in 366ml anhydrous methylene chloride, injected at room temperature borontrifluoride
Borate ether 280g adds 4 liters of methylene chloride dilutions after reacting 2h, is quenched with saturated sodium bicarbonate aqueous solution, then liquid separation, organic phase
It is dry with anhydrous sodium sulfate, obtain E5 (387g, yield 90%).
The preparation of compound E6:
Intermediate E 5 (86g) is dissolved in 860ml methanol, sodium methoxide 2.7g is added, acetic acid on the rocks after reaction 1h is stirred at room temperature
It is neutralized to neutrality, evaporated under reduced pressure, residue adds methylene chloride 1L, water 1L washing, and liquid separation is spin-dried for organic phase, and obtaining E6, (77.6g is received
Rate 100%.
The preparation of compound E7:
E6 (38.8g) is dissolved in 400ml acetonitrile, adds potassium hydroxide 12g, bromobenzyl 18g is added dropwise at 10 DEG C, add first after reacting 4h
Alcohol 50ml reflux 1h, then decompression is spin-dried for, and residue adds ethyl acetate 1L, water 1L extracting and washing, and organic phase decompression is spin-dried for slightly
Product 50g, crude product 40ml ethyl acetate, 120ml petroleum ether recrystallize to obtain E7 (45g, yield 94%).
The preparation of compound E8:
E6 (38.8g) is dissolved in 400ml acetonitrile, adds potassium hydroxide 23g, dimethyl suflfate 18.9g, reaction is added dropwise at 10 DEG C
Decompression is spin-dried for after 4h, and residue adds ethyl acetate 1, water 1L extracting and washing, and organic phase decompression is spin-dried for obtaining crude product 48g, and crude product is used
30ml ethyl acetate, 120ml petroleum ether recrystallize to obtain E8 (36.4g, yield 91%).
The preparation of compound E9:
E5 (98.5g) is dissolved in 430ml acetic acid, is added water 86g, 70 DEG C be stirred to react 1h after 55 DEG C of vacuum rotary steams it is dry, add
Water 1L, ethyl acetate 1L are adjusted to pH=7 with sodium carbonate, and then liquid separation obtains organic phase, and organic phase evaporated under reduced pressure obtains E9 crude product
80g, petroleum ether: ethyl acetate=1:1 column chromatographs to obtain sterling 70.5g, yield 90%.
The preparation of compound E10:
E7 (45g) is dissolved in 450ml acetic acid, is added water 90g, 70 DEG C be stirred to react 1h after 55 DEG C of vacuum rotary steams it is dry, add water
1L, ethyl acetate 1L are adjusted to pH=7 with sodium carbonate, and then liquid separation obtains organic phase, and organic phase evaporated under reduced pressure obtains E10 crude product
40g, petroleum ether: ethyl acetate=1:1 column chromatographs to obtain sterling 33.7g, yield 92%.
The preparation of compound E11:
E8 (36g) is dissolved in 450ml acetic acid, is added water 90g, 70 DEG C be stirred to react 1h after 55 DEG C of vacuum rotary steams it is dry, add water
1L, ethyl acetate 1L are adjusted to pH=7 with sodium carbonate, and then liquid separation obtains organic phase, and organic phase evaporated under reduced pressure obtains E11 crude product
38g, petroleum ether: ethyl acetate=1:1 column chromatographs to obtain sterling 26.7g, yield 95%.
The preparation of compound E12:
E9 (68.4g) and TEMPO 0.63g is added in methylene chloride 342ml and water 342ml, iodobenzene two is added at one time
Ethyl ester 86g is vigorously stirred room temperature reaction 45min, and then plus 30% sodium sulfite aqueous solution 200ml is quenched.Water phase acetic acid second
Ester extracts 5 times (500ml × 5), and organic phase is dry with anhydrous sodium sulfate, and evaporated under reduced pressure obtains E12 (57.6g, yield 80%).
The preparation of compound E13:
E10 (33g) and TEMPO 0.27g is added in methylene chloride 330ml and water 330ml, iodobenzene two is added at one time
Ethyl ester 37g is vigorously stirred lower room temperature reaction 45min, and then plus 30% sodium sulfite aqueous solution 100ml is quenched.Water phase acetic acid
Ethyl ester extracts 5 times (300ml × 5), and organic phase is dry with anhydrous sodium sulfate, and evaporated under reduced pressure obtains E13 (28g, yield 81%).
The preparation of compound E14:
E11 (26g) and TEMPO 0.27g is added in methylene chloride 260ml and water 260ml, iodobenzene two is added at one time
Ethyl ester 37g is vigorously stirred lower room temperature reaction 45min, and then plus 30% sodium sulfite aqueous solution 100ml is quenched.Water phase acetic acid
Ethyl ester extracts 5 times (300ml × 5), and organic phase is dry with anhydrous sodium sulfate, and evaporated under reduced pressure obtains E14 (25g, yield 88%).
The preparation of compound E15:
E12 (57.6g) is dissolved in acetonitrile 576ml, potassium carbonate 33.5g, iodomethane 34.5g is added, is stirred to react 8h, is reacted
Complete plus triethylamine 10ml is quenched, and saturated common salt water washing three times (200ml × 3), makes pH=7~8, and decompression is spin-dried for organic phase.Stone
Oily ether: ethyl acetate=2:1 column chromatographic purifying obtains 50g, yield 85%.
The preparation of compound E16:
E13 (28g) is dissolved in acetonitrile 280ml, potassium carbonate 9.6g and iodomethane 10g is added, is stirred to react 8h, has reacted and has added
Triethylamine 5ml is quenched, and saturated common salt water washing three times (100ml × 3), makes pH=7~8, and decompression is spin-dried for organic phase.Petroleum ether:
Ethyl acetate=2:1 column chromatographic purifying, obtains 25g, yield 87%.
The preparation of compound E17:
E14 (25g) is dissolved in acetonitrile 250ml, potassium carbonate 10.5g and iodomethane 11g is added, is stirred to react 8h, has reacted
Triethylamine 5ml is added to be quenched, saturated common salt water washing three times (100ml × 3), makes pH=7~8, and decompression is spin-dried for organic phase.Petroleum
Ether: ethyl acetate=2:1 column chromatographic purifying obtains 23.5g, yield 90%.
The preparation of Part III monosaccharide F ring and H ring
Synthetic route:
A) sulfuric acid, benzaldehyde, DMF, b) benzyl bromine, potassium hydroxide, acetonitrile, c) triethylsilane, boron trifluoride ether, dichloro
Methane, d) acetic acid, sulfuric acid, water, e) aceticanhydride, triethylamine, DMAP, ethyl acetate, f) piperidines, tetrahydrofuran, g) Tritox,
DBU, methylene chloride, h) acetic acid, water, i) -40 DEG C, chlorobenzoyl chloride, pyridine
The preparation of compound F1 (H1)-F3 (H3): referring to Bioorganic&Medicinal Chemistry Letters,
2009,19(14),3875-3879。
F3 (464g): being added the mixed solution of acetic acid 2.3kg and water 464g by the preparation of compound F4, and sulfuric acid 46g is added,
1h is reacted under the conditions of 100 DEG C.Evaporated under reduced pressure has been reacted, F4 (427.5g, yield 95%) is obtained.
The preparation of compound F5: ethyl acetate dissolution is added in F4 (400g), triethylamine 200ml is added, aceticanhydride is added dropwise
200ml adds saturated aqueous sodium carbonate to neutralize after reacting 1h, and ethyl acetate extraction, evaporated under reduced pressure obtains F5 (475g, yield
100%).1H NMR(300MHz,CDCl3) δ 7.35-7.29 (m, 15H), 6.0 (d, J=3.6Hz, 1H), 4.9 (dd, J=9.2,
3.4Hz,1H),4.7-4.6(m,6H),4.4(m,1H),4.27-4.18(m,2H),3.6(d,1H),3.4(d,1H),2.19(s,
3H),2.14(s,3H).MS(ESI):557.3[M+Na]+
The preparation of compound F6: tetrahydrofuran dissolution is added in F5 (450g), piperidinyl-1 43g is added, adds acetic acid after reacting 6h
It neutralizes, vacuum rotary steam, residue ethyl acetate is dissolved, washed, and evaporated under reduced pressure organic phase obtains 373g, yield 90%.
The preparation of compound F7: being added methylene chloride dissolution for F6 (350g), Tritox 400g and DBU 10g be added,
It is spin-dried for after reaction 1h, column chromatography obtains F7 (406g, yield 90%).
The preparation of compound H4: F2 (463g) is added to the mixed solution of acetic acid 2.3kg and water 464g, adds sulfuric acid
46g, 100 DEG C of reaction 1h.Evaporated under reduced pressure has been reacted, H4 (307g, yield 82%) is obtained.
The preparation of compound H5: pyridinium dissolution is added in H4 (187g), chlorobenzoyl chloride is slowly added dropwise under -40 DEG C of coolings
141g, adds saturated aqueous solution of sodium bicarbonate that faint yellow solid is precipitated, washs after reacting 2h, and drying obtains H5 (203.3g, yield
85%).
Part IV: the synthesis of monosaccharide G ring
Synthetic route:
A) dimethyl suflfate, KOH, tetrahydrofuran, b) phosphomolybdic acid, silica gel, water, acetonitrile, c) dipotassium hydrogen phosphate trihydrate, phosphorus
Acid dihydride potassium, water, TEMPO, NaClO2, KBr, TCCA, d) iodomethane, potassium carbonate, acetonitrile, e) trifluoromethanesulfanhydride anhydride, pyridine, f)
Sodium trifluoroacetate, DMSO, water, g) trifluoracetic acid, water, h) aceticanhydride, triethylamine, DMAP, ethyl acetate, j) trifluoromethanesulfonic acid neodymium,
K) Tritox, DBU, methylene chloride
The preparation of compound G1: referring to the synthesis of E1.
The preparation of compound G2: being dissolved in acetonitrile 9.72kg for G1 (1.5kg), add silica gel 0.36kg, phosphomolybdic acid 0.036kg,
16h is stirred at room temperature in water 0.8kg, filters off silica gel, is neutralized to pH=7, filtrate decompression revolving with aqueous sodium carbonate, residue adds
Water, ethyl acetate extraction, filtrate decompression are evaporated to obtain G2 (1.15kg, yield 90%).
The preparation of compound G3: being dissolved in tetrahydrofuran 10L for G2 (1.15kg), be added dipotassium hydrogen phosphate trihydrate 500g,
Potassium dihydrogen phosphate 500g, water 1.5kg, TEMPO 20g, NaClO2300g, KBr 50g and TCCA 500g add nothing after reacting 1h
Water sodium sulfite is quenched, and then decompression is spin-dried for, and residue is washed with ethyl acetate, and organic phase decompression is spin-dried for obtaining G3 (1.1kg, receipts
Rate 90%).
The preparation of compound G4: G3 (1.1kg) is dissolved in acetonitrile 6.0kg, 1.22kg potassium carbonate and iodomethane is added
Reaction 8h is stirred at room temperature in 0.73kg, after completion of the reaction, triethylamine is added, iodomethane is quenched and filters, filtrate decompression is spin-dried for obtaining G4
(929g, yield 80%).
The preparation of compound G5: pyridine 1L and acetonitrile 5L is added in G4 (885g), trifluoromethanesulfanhydride anhydride 950g is added dropwise, is added dropwise
Reaction 1h is finished, then elutriation crystalline substance on the rocks filters, and filter cake is dissolved with methylene chloride, and then methylene chloride is mutually washed with water, dries, and subtracts
Pressure is spin-dried for obtaining G5 (1280g, yield 96%).
The preparation of compound G6: G5 (1280g) and sodium trifluoroacetate 1kg are dissolved in DMSO 10L and water 1L, stir 8h
Afterwards plus ethyl acetate extraction, decompression are spin-dried for obtaining G6 (808g, yield 95%).
The preparation of compound G7: trifluoracetic acid 2L is added in G6 (808g), water 200ml is added, depressurizes and is spin-dried for after stirring 1h
It obtains G7 (547g, yield 80%).
The preparation of compound G8: methylene chloride 5L dissolution is added in G7 (547g), adds DMAP 50g and triethylamine
Then aceticanhydride 1.15kg is added dropwise at 0 DEG C by 1.15kg, react after 0.5h plus water washing, organic phase decompression are spin-dried for, obtaining G8, (815g is received
Rate 95%).
The preparation of compound G9: by G8 (810g) be added methanol 12L, be added trifluoromethanesulfonic acid neodymium 60g, stir 12h after it is dense
Contracting, column chromatograph to obtain G9 (605g, yield 85%).
The preparation of compound G10: being added methylene chloride 6kg for G9 (605g), Tritox 1.2kg and DBU 45g be added,
It is spin-dried for after reaction 1h, column chromatography obtains G10 (801g, yield 90%).
The synthesis of Part V disaccharides DE
Synthetic route:
A) TMSOTf, methylene chloride, b) ammonium ceric nitrate, c) Tritox, DBU, methylene chloride
The preparation of compound DE1: it takes D3 (50g) and E15 (30g) to be dissolved in anhydrous methylene chloride, is added dropwise under nitrogen protection
After TMSOTf1g, -20 DEG C of reaction 1h plus triethylamine is quenched, and decompression is spin-dried for, and column chromatography obtains DE1 αisomer 51g, yield 85%.
The preparation of compound DE2: it takes D3 (55g) and E16 (30g) to be dissolved in anhydrous methylene chloride, is added dropwise under nitrogen protection
After TMSOTf1g, -20 DEG C of reaction 1h plus triethylamine is quenched, and decompression is spin-dried for, and column chromatography obtains DE2 αisomer 46.9g, yield
83%.
The preparation of compound DE3: it takes D3 (50g) and E17 (30g) to be dissolved in anhydrous methylene chloride, is added dropwise under nitrogen protection
After TMSOTf1g, -20 DEG C of reaction 1h plus triethylamine is quenched, and decompression is spin-dried for, and column chromatography obtains DE2 αisomer 50.8g, yield
82%.
The preparation of compound DE4: by DE1 (40g) be dissolved in acetonitrile 0.4L toluene 0.2L water 0.2L mixed solution, be added
Ammonium ceric nitrate 50g, ethyl acetate extracts after reacting at room temperature 1h, and decompression is spin-dried for obtaining DE4 (30.9g, yield 90%).
1H NMR(300MHz,CDCl3) δ 7.35-7.29 (m, 10H), 6.29 (d, J=3.6Hz, 1H), 5.6 (d, J=
3.4Hz, 1H), 4.97 (dd, J=9.2,3.4Hz, 1H), 4.65-4.54 (m, 5H), 4.32-4.18 (m, 3H), 4.00 (m,
1H),3.80(m,2H),3.70(s,3H),3.65–3.1(m,12H),2.02(s,3H).MS(ESI):657.3[M+Na]+
The preparation of compound DE5: by DE2 (40g) be dissolved in acetonitrile 0.4L toluene 0.2L water 0.2L mixed solution, be added
Ammonium ceric nitrate 45g, ethyl acetate extracts after reacting at room temperature 1h, and decompression is spin-dried for obtaining DE5 (31.5g, yield 91%)
1H NMR(300MHz,CDCl3) δ 7.36-7.26 (m, 15H), 5.9 (d, J=3.6Hz, 1H), 5.6 (dd, J=
9.2,3.4Hz,1H),4.65–4.54(m,7H),4.32(s,1H),4.23(m,1H),4.00(m,1H),3.80(m,3H),
3.70(s,3H),3.65–3.1(m,10H).MS(ESI):705.3[M+Na]+
The preparation of compound DE6: DE3 40g, add acetonitrile 0.4L toluene 0.2L water 0.2L, be added ammonium ceric nitrate 50g, room
Ethyl acetate extracts after temperature reaction 1h, and decompression is spin-dried for obtaining DE6 (32g, yield 94%)1H NMR(300MHz,CDCl3)δ7.34-
7.29 (m, 10H), 5.90 (d, J=3.6Hz, 1H), 5.60 (d, J=3.6Hz, 1H), 4.65-4.61 (m, 5H), 4.32 (s,
1H),4.23(d,1H),4.00(m,1H),3.80-3.77(m,3H),3.70(s,3H),3.61-3.60(m,2H),3.50(m,
1H),3.41(m,12H).MS(ESI):629.3[M+Na]+
The preparation of compound DE7: being added methylene chloride 0.6kg for DE4 (30g), Tritox 40g and DBU 4g be added,
It is spin-dried for after reaction 1h, column chromatography, obtains DE7 (29.5g, yield 80%).
The preparation of compound DE8: being added methylene chloride 0.6kg for DE5 (30g), Tritox 40g and DBU 4g be added,
It is spin-dried for after reaction 1h, column chromatography, obtains DE8 (29.8g, yield 82%).
The preparation of compound DE9: being added methylene chloride 0.6kg for DE4 (30g), Tritox 40g and DBU 4g be added,
It is spin-dried for after reaction 1h, column chromatography, obtains DE9 (31.5g, yield 85%).
The synthesis of Part VI trisaccharide FGH
Synthetic route:
A) TMSOTf, methylene chloride, b) potassium carbonate, methanol, c) TMSOTf, methylene chloride, d) tribromo-acetyl imines benzyl ester,
Trifluoromethanesulfonic acid, e) potassium carbonate, methanol
The preparation of compound GH1: G10 (631g) and H3 (465g) plus anhydrous methylene chloride are dissolved, and are added dropwise at 0 DEG C
TMSOTf23g adds triethylamine to be quenched after reacting 1h, and column chromatography obtains GH1 (677g, yield 90%).
The preparation of compound GH2: G10 (631g) and H5 (478g) plus anhydrous methylene chloride are dissolved, and are added dropwise at 0 DEG C
TMSOTf23g adds triethylamine to be quenched after reacting 1h, and column chromatography obtains GH2 (659g, yield 86%).
The preparation of compound GH3: GH1 (600g) and potassium carbonate (400g) plus anhydrous methanol are dissolved, and react 6h at 25 DEG C
Afterwards plus water, ethyl acetate extract, and are spin-dried for, and column chromatography obtains GH3 (432g, yield 81%).
The preparation of compound GH4: GH2 (600g) and potassium carbonate (400g) plus anhydrous methanol are dissolved, and react 6h at 25 DEG C
Afterwards plus water, ethyl acetate extract, and are spin-dried for, and column chromatography obtains GH4 (438g, yield 82%).
The preparation of compound FGH1: GH3 (400g) and F7 (520g) plus anhydrous methylene chloride are dissolved, and are added dropwise at 0 DEG C
TMSOTf15g adds triethylamine to be quenched after reacting 1h, and column chromatography obtains FGH1 (561g, yield 82%).
The preparation of compound FGH2: GH4 (400g) and F7 (530g) plus anhydrous methylene chloride are dissolved, and are added dropwise at 0 DEG C
TMSOTf15g adds triethylamine to be quenched after reacting 1h, and column chromatography obtains FGH2 (439g, yield 75%).
The preparation of compound FGH3: FGH1 (400g) and tribromo-acetyl imines benzyl ester (125g) plus anhydrous methylene chloride is molten
It solves, trifluoromethanesulfonic acid 10g is added dropwise at 0 DEG C, add triethylamine to be quenched after reacting 1h, column chromatography obtains FGH3 (388g, yield 90%).
The preparation of compound FGH4: FGH2 (400g) and tribromo-acetyl imines benzyl ester (125g) plus anhydrous methylene chloride are dissolved, at 0 DEG C
Trifluoromethanesulfonic acid 10g is added dropwise, adds triethylamine to be quenched after reacting 1h, column chromatography obtains FGH4 (397g, yield 92%).Compound
The preparation of FGH5: FGH3 (300g) and potassium carbonate (400g) plus anhydrous methanol are dissolved, and add water, acetic acid second after 6h is reacted at 25 DEG C
Ester extraction, is spin-dried for, and column chromatography obtains FGH5 (243g, yield 84%).
1H NMR(300MHz,CDCl3)δ7.32-7.30(m,35H),5.60(m,3H),4.88(s,1H),4.64–4.62
(m,15H),4.49(m,1H),4.24-4.23(m,1H),4.00(m,1H),3.80-3.79(m,7H),3.70(m,2H),
3.61-3.60(s,3H),3.50(d,1H),3.41-3.36(m,8H).MS(ESI):1213.5[M+Na]+
The preparation of compound FGH6: FGH4 (300g) and potassium carbonate (400g) plus anhydrous methanol are dissolved, and are reacted at 25 DEG C
After 6h plus water, ethyl acetate extract, and are spin-dried for, and column chromatography obtains FGH6 (249g, yield 86%).
1H NMR(300MHz,CDCl3)δ8.06-7.55(m,5H),7.32-7.30(m,30H),5.60(m,3H),4.88
(s,1H),4.64–4.62(m,13H),4.49(m,1H),4.24-4.23(m,2H),3.80-3.77(m,6H),3.61-3.60
(m,2H),3.50(d,1H),3.41-3.40(d,6H).MS(ESI):1227.5[M+Na]+
The preparation of 1 pentasaccharides III-1 (sodium salt) of embodiment
Synthetic route:
A) TMSOTf, methylene chloride, b) palladium carbon, hydrogen, c) sulfur trioxide triethylamine salt, DMF, sodium bicarbonate, dowex-
50x4, d) 4N NaOH, sephadex g25
The preparation of compound DEFGH10: DE5 (29g) and FGH6 (45g) plus anhydrous methylene chloride are dissolved, and are added dropwise at 0 DEG C
TMSOTf 1g adds triethylamine to be quenched after reacting 1h, and column chromatography obtains DEFGH10 (59g, yield 87%).1H NMR(300MHz,
CDCl3) δ 8.05-7.55 (m, 5H), 7.34-7.29 (m, 40H), 5.99 (d, 1H), 5.60 (m, 4H), 5.17 (dd, J=9.0,
3.4Hz,1H),4.64–4.62(m,18H),4.49-4.19(m,5H),4.00(m,2H),3.80-3.77(m,8H),3.70(s,
6H),3.61(d,2H),3.50(d,3H),3.41-3.40(m,15H),3.36(m,2H),2.02(s,3H).MS(ESI):
1843.8[M+Na]+
The preparation of compound DEFGH11: DEFGH10 (59g) is dissolved in anhydrous methanol, 10% palladium carbon, hydrogen normal pressure is added
Filtering is spin-dried for obtaining DEFGH11 (34.2g, yield 96%) after reductase 12 4h.1H NMR(300MHz,CDCl3)δ8.05-7.55(m,
5H), 5.99 (d, J=3.6Hz, 1H), 5.40 (m, 4H), 5.17 (dd, J=9.0,3.4Hz, 1H), 4.77 (d, 4H), 4.71
(d,2H),4.64–4.62(m,2H),4.49-4.19(m,6H),4.10(m,2H),3.94-3.80(m,8H),3.70(d,6H),
3.60-3.50(m,7H),3.41-3.40(m,15H),3.30(m,2H),2.02(s,3H).MS(ESI):1123.4[M+Na]+
The preparation of compound DEFGH12: n,N-Dimethylformamide dissolution is added in DEFGH11 (5.8g), is added excessive
Sulfur trioxide triethylamine salt, 50 DEG C of reactions add saturated sodium bicarbonate aqueous solution to be quenched, are concentrated under reduced pressure afterwards for 24 hours.Gel column
(Sephadex G-25) separation, eluent (water/acetonitrile=7/1, v/v).Sodium form ion exchange resin column (DOWEX-50Na),
Pure water elution, is concentrated under reduced pressure to give colorless solid DEFGH12 (9.0g, yield 89%).1H NMR(300MHz,CDCl3)δ8.05-
7.55 (m, 5H), 5.99 (d, J=3.6Hz, 1H), 5.54 (m, 4H), 5.30-5.17 (m, 7H), 4.64-4.62 (m, 2H),
4.49-4.19(m,8H),3.96-3.94(m,4H),3.74(m,2H),3.70(d,6H),3.50-3.44(m,3H),3.41-
3.40(m,15H),2.02(s,3H).MS(ESI):935.0[(M-2Na)/2]-
The preparation of pentasaccharides III-1 (sodium salt): 4N NaOH aqueous solution is added in DEFGH12 (5.2g), after room temperature reaction for 24 hours
It is concentrated under reduced pressure.Gel column (Sephadex G-25) separation, eluent (water/acetonitrile=4/1, v/v).Sodium form ion exchange resin
Column (DOWEX-50Na), pure water elution, is concentrated under reduced pressure to give colorless solid (4.1g, yield 85%).
1H NMR(300MHz,CDCl3)δ5.54(m,4H),5.40(m,1H),5.30(m,2H),5.25-5.24(m,4H),
4.77(m,1H),4.64–4.62(m,2H),4.23-4.21(m,5H),4.10(m,1H),3.96-3.94(m,5H),3.80-
3.74(m,3H),3.60(m,2H),3.51-3.40(m,19H).MS(ESI):870.0[(M-2Na)/2]-
The preparation of 2 pentasaccharides III-2 (sodium salt) of embodiment
Synthetic route:
It referring to the method for embodiment 1, reacts using DE9 and FGH6 as raw material, then through hydrogenolysis removing benzyl, generation
Target compound is made in sulfating reaction, hydrolysis.
Compound DEFGH20:
1H NMR(300MHz,CDCl3)δ8.05-7.55(m,5H),7.34-7.29(m,45H),5.60(m,5H),4.64–
4.62(m,20H),4.49-4.19(m,3H),4.00(m,2H),3.80-3.77(m,10H),3.70(s,6H),3.61(d,
2H),3.50(d,3H),3.41-3.40(m,15H),3.36(m,2H),2.02(s,3H).MS(ESI):1891.8[M+Na]+
Compound DEFGH21:
1H NMR(300MHz,CDCl3)δ8.05-7.55(m,5H),5.40(m,5H),4.77(d,5H),4.71(d,2H),
4.64–4.62(m,2H),4.49-4.19(m,5H),4.10(m,3H),3.94-3.80(m,8H),3.70(d,6H),3.60-
3.50(m,8H),3.41-3.40(m,15H),3.30(m,2H).MS(ESI):1081.4[M+Na]+
Compound DEFGH22:
1H NMR(300MHz,CDCl3)δ8.05-7.55(m,5H),5.54(m,5H),5.30-5.24(m,7H),,4.64–
4.62(m,2H),4.49-4.19(m,7H),3.96-3.94(m,4H),3.74(m,3H),3.70(d,6H),3.50-3.44(m,
3H),3.41-3.40(m,15H).MS(ESI):964.9[(M-2Na)/2]-
Pentasaccharides III-2 (sodium salt):
1H NMR(300MHz,CDCl3)δ5.54(m,5H),5.40(m,1H),5.30(m,3H),5.25-5.24(m,3H),
4.64–4.62(m,2H),4.23-4.21(m,4H),3.96-3.94(m,5H),3.80-3.74(m,4H),3.60-3.44(m,
5H),3.41-3.40(m,15H).MS(ESI):920.9[(M-2Na)/2]-
3 pentasaccharides III-3 (R of embodiment1For methyl, sodium salt) preparation
Synthetic route:
It referring to the method for embodiment 1, reacts using DE6 and FGH6 as raw material, then removes benzyl, sulfuric acid through hydrogenolysis
Change reaction, target compound is made in hydrolysis.
Compound DEFGH30:
1H NMR(300MHz,CDCl3)δ8.05-7.55(m,5H),7.34-7.29(m,40H),4.64–4.62(m,
18H),4.49-4.19(m,4H),4.00(m,2H),3.80-3.77(m,10H),3.70(s,6H),3.61(d,2H),3.50
(d,3H),3.41-3.40(m,18H),3.36(m,2H).MS(ESI):1815.8[M+Na]+
Compound DEFGH31:
1H NMR(300MHz,CDCl3)δ8.05-7.55(m,5H),5.60(d,1H),5.40(m,4H),4.77(d,4H),
4.71(d,2H),4.64–4.62(m,2H),4.49-4.19(m,5H),4.10(m,2H),3.94-3.80(m,10H),3.70
(d,6H),3.60-3.50(m,7H),3.41-3.40(m,18H),3.30(m,2H).MS(ESI):1095.4[M+Na]+
Compound DEFGH32:
1H NMR(300MHz,CDCl3)δ8.05-7.55(m,5H),5.60(d,1H),5.54(m,4H),5.30-5.24
(m,6H),4.64–4.62(m,2H),4.49-4.19(m,7H),3.96-3.94(m,4H),3.80-3.70(m,10H),3.50-
3.44(m,3H),3.41-3.40(m,18H).MS(ESI):921.0[(M-2Na)/2]-
Pentasaccharides III-3 (sodium salt):
1H NMR(300MHz,CDCl3)δ5.60-5.54(m,5H),5.30-5.24(m,6H),4.64–4.62(m,2H),
4.23-4.21(m,4H),3.96-3.94(m,5H),3.80-3.74(m,5H),3.60-3.44(m,5H),3.41-3.40(m,
18H).MS(ESI):876.9[(M-2Na)/2]-
The preparation of 4 pentasaccharides III-4 (sodium salt) of embodiment
Synthetic route:
It referring to the method for embodiment 1, reacts using DE5 and FGH5 as raw material, then through hydrogenolysis removing benzyl, generation
Target compound is made in sulfating reaction, hydrolysis.
Compound DEFGH40:
1H NMR(300MHz,CDCl3)δ7.34-7.29(m,45H),5.60(d,1H),5.40(m,4H),5.17(m,
1H),4.64–4.62(m,20H),4.49-4.19(m,3H),4.00(m,3H),3.80-3.77(m,8H),3.70(s,6H),
3.61(d,3H),3.50(d,3H),3.41-3.40(m,15H),3.36(m,3H).MS(ESI):18298[M+Na]+
Compound DEFGH41:
1H NMR(300MHz,CDCl3)δ5.99(m,1H),5.60(d,1H),5.40(m,4H),5.17(m,1H),4.77
(d,4H),4.71(d,2H),4.64–4.62(m,2H),4.49-4.19(m,3H),4.10(m,2H),3.94-3.80(m,
10H),3.70(d,6H),3.60-3.50(m,7H),3.41-3.40(m,15H),3.30(m,2H),2.02(s,3H).MS
(ESI):1019.4[M+Na]+
Compound DEFGH42:
1H NMR(300MHz,CDCl3)δ5.99(d,1H),5.54(m,4H),5.30-5.24(m,6H),5.17(m,1H),
4.64–4.62(m,2H),4.49-4.19(m,6H),3.96-3.94(m,6H),3.74(m,2H),3.70(m,6H),3.50-
3.44(m,3H),3.41-3.40(m,15H).MS(ESI):933.9[(M-2Na)/2]-
Pentasaccharides III-4 (sodium salt):
1H NMR(300MHz,CDCl3)δ5.60-5.54(m,4H),5.40(m,1H),5.30-5.24(m,6H),4.77
(s,1H),4.64–4.62(m,2H),4.23-4.21(m,5H),4.10(m,1H),3.96-3.94(m,6H),3.74(m,2H),
3.60-3.44(m,4H),3.41-3.40(m,15H).MS(ESI):920.9[(M-2Na)/2]-
5 biological test of embodiment
Five sugar compounds of the invention biological activity test can using well known to a person skilled in the art method into
Row measurement.It is understood that following test methods are not limited in any way the scope of the present invention.
1.AT III combination KDValue measurement
Using Fluorescence Method, Perkin Elmer LS-50 type sepectrophotofluorometer is used；Excite λ 280nm, transmitting
λ338nm；Constant temperature sample room is equipped with to continuously stir at 37 DEG C.Respectively by five sugar compounds of target made from Examples 1 to 4
2mL Tris-HCl buffer (0.01M, pH value 7,0.15M NaCl and 5-60nM people AT- III) test tube is added.
The ratio of III-five saccharide complex of AT- is reacted with the calculating of concentration by 1:1 to measure, dissociation constant KDMeasurement adopt
It is analyzed with Scatchard.As a result average value ± SEM, n=3.
2. the measurement of anticoagulin xa activity
Five sugar compounds of the invention inhibit factor Xa by activation antithrombase AT III.Anti-Xa activity measurement
Using anti-Xa Amidolytic AT III method (i.e. chromogenic substrate method).At 37 DEG C, factor Xa is (in 20mM Tris/ maleic acid
In salt buffer, concentration is 7.5nkat/ milliliters, pH=7.4, NaCl 150mM；100 μ L), AT III is (in 20mM Tris/ horse
Come in phthalate buffer, concentration is 0.5 units per ml, pH=7.4, NaCl 150mM；100 μ L) and pentasaccharides of the invention (
In 20mM Tris/ maleate buffers, concentration is 0.5 units per ml, pH=7.4, NaCl 150mM；100 μ L) culture 2
Minute.Then S-2222 culture medium (Bz- lle-Glu-glycine-arginine pNA is added；1mM is in 50mM Tris-
In HCl buffer, pH=8.4,175mM NaCl, EDTA 27.5mM；100 μ L) measure remaining factor Xa.2 minutes
The acetic acid aqueous solution (100 μ L) that 50% is added afterwards stops reaction, and absorbance is measured at 405nm.Then the percentage of inhibition is calculated
Than formula: inhibiting %=100 × (A of control buffer405The A of sample405The A of)/control buffer405；With the mark calibrated
Standard relatively, uses the activity of 2015 software of Excel measurement compound.It as a result is average value ± SEM, n=3.
3. half-life period (T1/2) measurement
After intravenous administration, the pharmacokinetics of five sugar compounds of the invention is carried out in male Wistar rat
Research, dosage 100nmol/Kg.Blood sample is acquired at multiple time points after administration, by the blood sample of 9 volumes and 1 volume
The mixing of 0.129M sodium citrate solution and immediately ice bath cooling make sample receive 3000Xg at low temperature and are centrifuged 10 minutes, and-
It is frozen at 20 DEG C.The concentration of compound in every milliliter of blood plasma is measured by coagulation factor xa activity.For each compound, by dense
It spends the curve relative to the time and removes half-life period to calculate.
As a result as shown in the table:
| Compound | AT III combination KDIt is worth (nM) | Anti-Xa factor activity (units/mg) | Half-life period (T1/2,h) |
| III-1 | 24±1 | 850±25 | 1.4±0.1 |
| III-2 | 22±3 | 870±26 | 1.2±0.1 |
| III-3 | 20±2 | 920±30 | 1.3±0.1 |
| III-4 | 24±1 | 1190±35 | 1.5±0.1 |
| Fondaparinux sodium | 58±3 | 850±27 | 0.7±0.1 |
Upper table data show that five sugar compounds of the invention have potent Anti-Xa activity, compared with Fondaparinux sodium, this
The Anti-Xa activity of invention compound is its 1~1.4 times；And the half-life period of five sugar compounds of the invention is more compared with Fondaparinux sodium
It is long.
4. bleeding influences test
Male Wistar rat (weight 18-22g) is randomly divided into 4 test groups, blank group and control group, every group 12
Only.4 test groups are injected intravenously pentasaccharides III-1, III-2, III-3, III-4 respectively, and control group is injected intravenously sulphur and reaches liver
Last of the ten Heavenly stems sodium, blank group are injected intravenously physiological saline.Every group is further divided into four groups, respectively with 1mg/Kg, 3mg/Kg, 10mg/kg
It is administered with the dosage of 15mg/kg, five sugar compounds of the invention are in terms of its sodium salt.After 1h is administered, the tail of rat is cut about
5mm simultaneously immerses in 40mL distilled water, and constant temperature stirs 90min at 37 DEG C.Using hemoglobin in spectrophotometric method measurement water
Amount, Detection wavelength 540nm prepares standard curve, calculates amount of bleeding.Results are averaged ± SD, n=3.
For upper table the results show that under high dosage, the influence of five sugar compounds of the invention to hemorrhagic tendency is significant
Less than positive control Fondaparinux sodium.
Claims (15)
1. five sugar compounds of synthesis and its salt of ionic species shown in formula III,
Wherein, work as R1When for H, R2For H or SO3 -；
Work as R1For SO3 -Or when C1~C4 alkyl, R2For H.
2. five sugar compounds according to claim 1 or its acid or its salt, which is characterized in that have the following structure:
Wherein, R1For C1~C4 alkyl.
3. five sugar compounds according to claim 2 or its acid or its salt, which is characterized in that R1For methyl.
4. five sugar compounds according to claim 1-3 or its acid or its salt, which is characterized in that the salt is
Sodium salt, sylvite, calcium salt or ammonium salt.
5. the method for preparing five sugar compounds as claimed in claim 1 or 2 or its acid or its salt, which is characterized in that include following step
It is rapid:
(a) trisaccharide shown in disaccharides shown in formula DE and formula FGH is reacted preparation formula DEFGH0,
Wherein, R1Selected from Ac, Bn, C1~C4 alkyl, R2Selected from Bn, Bz.
(b) by compound shown in formula DEFGH0 remove benzyl, preparation formula DEFGH1,
Wherein, RaFor Ac, H, C1~C4 alkyl, RbFor H, Bz；
(c) sulfating reaction preparation formula DEFGH2 is occurred into for compound shown in formula DEFGH1,
Wherein, RcFor Ac, SO3 -, C1~C4 alkyl, RdFor SO3 -, Bz；
(d) formula DEFGH2 is hydrolyzed, prepares pentasaccharides III-1~III-4.
6. the compound being shown below
Wherein, R1Selected from Ac, Bn, C1~C4 alkyl.
7. the compound being shown below:
Wherein, R2Selected from Bn, Bz.
8. the compound being shown below:
Wherein, R1Selected from Ac, Bn, C1~C4 alkyl, R2Selected from Bn, Bz.
9. the compound being shown below:
Wherein, RaFor Ac, H, C1~C4 alkyl, RbFor H, Bz.
10. the compound being shown below
Wherein, RcFor Ac, SO3 -, C1~C4 alkyl, RdFor SO3 -, Bz.
11. pharmaceutical composition, it includes as active constituent compound according to any one of claims 1 to 4, its acid or
Its salt and optional pharmaceutically acceptable auxiliary material.
12. pharmaceutical composition according to claim 11, the active constituent containing 0.1mg~100mg.
13. described in compound according to any one of claims 1 to 4, its acid or any one of its salt or claim 11-12
Pharmaceutical composition in preparing drug for preventing and/or treating disease relevant to blood coagulation disorder and morbid state
Purposes, the disease and morbid state are preferably Deep vain thrombosis, thrombophlebitis, are drawn by thrombosis or embolism
Artery occlusion, Postoperative Intravenous thrombosis or embolism, pulmonary embolism and the non-ST elevation acute myocardial infraction risen.
14. the method for preventing and/or treating relevant to blood coagulation disorder disease and morbid state comprising Xiang Youxu
The individual wanted gives compound according to any one of claims 1 to 4, any in its acid or its salt or claim 11-12
Pharmaceutical composition described in, the disease and morbid state are preferably Deep vain thrombosis, thrombophlebitis, by thrombus
Artery occlusion caused by formation or embolism, Postoperative Intravenous thrombosis or embolism, pulmonary embolism and non-ST elevation acute myocardial infraction.
15. appointing for preventing and/or treating in disease relevant to blood coagulation disorder and the Claims 1 to 4 of morbid state
Compound described in one, its acid or any one of its salt or claim 11-12 described in pharmaceutical composition, the disease and
Morbid state is preferably Deep vain thrombosis, thrombophlebitis, the artery occlusion as caused by thrombosis or embolism, postoperative
Venous thronbosis or embolism, pulmonary embolism and non-ST elevation acute myocardial infraction.
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| WO2020114418A1 (en) * | 2018-12-05 | 2020-06-11 | 南京正大天晴制药有限公司 | Anticoagulant pentasaccharide compound, composition, and preparation method and medical use thereof |
| CN114957354A (en) * | 2021-02-25 | 2022-08-30 | 南开大学 | Heparin pentasaccharide structural compound |
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| WO2020114418A1 (en) * | 2018-12-05 | 2020-06-11 | 南京正大天晴制药有限公司 | Anticoagulant pentasaccharide compound, composition, and preparation method and medical use thereof |
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| CN114957354B (en) * | 2021-02-25 | 2023-11-24 | 南开大学 | Heparin pentasaccharide structure compound |
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| CN109134555B (en) | 2021-09-28 |
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