CN100360500C - N,N'-biacetylcysteine-diarginine salt isomer and its uses - Google Patents
N,N'-biacetylcysteine-diarginine salt isomer and its uses Download PDFInfo
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- CN100360500C CN100360500C CNB2005100411014A CN200510041101A CN100360500C CN 100360500 C CN100360500 C CN 100360500C CN B2005100411014 A CNB2005100411014 A CN B2005100411014A CN 200510041101 A CN200510041101 A CN 200510041101A CN 100360500 C CN100360500 C CN 100360500C
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Abstract
The present invention relates to configuration isomers of N, N'-diacetylcystine-diarginine salt, a preparation method for the configuration isomers and a medical use of the configuration isomers, and relates to a pharmaceutical composition using a preferable configuration isomer as an active component. A configuration isomer of which the cystine in molecules is in an L shape or a meson shape is preferably selected from the configuration isomers of the N, N'-diacetylcystine-diarginine salt, and the configuration of arginine in the molecules is formed by a configuration isomer containing at least one L-shaped arginine. The present invention is used for preparing antitumor medicine, medicine for treating immune deficiency diseases and autoimmune diseases and medicine for treating infectious diseases or treating hepatic diseases, hepatic injury and hepatic failure.
Description
Technical field
The present invention relates to have the N of immunoregulatory activity, the arginic acid salt of N '-diacetyl Gelucystine contains N, pharmaceutical composition, its preparation method and the medicinal use of N '-diacetyl Gelucystine two arginic acid salt configurational isomers.
Technical background
N-acetylcystein is a kind of known amino acid derivative that contains sulfydryl, has multiple biological activity, for example as mucolytic agent, is used for respiratory system disease; Also be studied and using aspect reduced glutathion reduction and the oxidative stress; Still can be used for the acute hepatic failure that drug intoxication causes.Clinical study in recent years proves that the liver failure that each Hepatitis virus is caused also has good efficacy.The pharmacological mechanism of N-acetylcystein is various, comprises strong antioxidant action, stimulates gsh synthetic, to influence of NO production and its synthase activity or the like.
N, N '-diacetyl Gelucystine are the dimer of N-acetylcystein with disulfide-bonded, also are a kind of known compound.Swedish patent application SE9002067-8 has found N, and N '-diacetyl Gelucystine has potent immunostimulation, can stimulate I type immediate allergy, suppresses the delayed allergy of IV type.Can also prove N in addition, N '-diacetyl Gelucystine can obviously improve the easypro blood function of damaged endotheliocyte, but does not influence the easypro blood function of normal endothelial cell.
Chinese patent " N; the organic salt of N '-diacetyl Gelucystine " (application number 93104963.6) discloses N, the salt that N '-diacetyl Gelucystine and some organic basess form can be avoided free N, and the water absorbability of N '-diacetyl Gelucystine helps preparing the preparation that meets drug quality control requirement.
Chinese patent application " new compound N, N '-diacetyl Gelucystine arginic acid salt and its production and use " (application number 02136216.5) discloses N, N '-diacetyl Gelucystine arginic acid salt and preparation method thereof and medicinal use.
N, the structural formula of N '-diacetyl Gelucystine two arginic acid salts is as follows:
R wherein
+Be arginic positively charged ion.
N, N '-diacetyl Gelucystine two arginic acid salts are white crystalline powder, with the free N of buttery, N '-diacetyl Gelucystine is compared, and has that water absorbability is little, the advantage of good stability.N with preparation in the Chinese patent " N; the organic salt of N '-diacetyl Gelucystine " (application number 93104953.6), N '-diacetyl Gelucystine lysine salt is compared, arginine and N, not only have the good physics that is suitable for preparation, chemical property behind N '-diacetyl Gelucystine salify, have more outstanding synergy, improve the curative effect of treatment hepatopathy, fulminant hepatic failure.But in this piece document, this compound structure type isomer is not selected.
Gelucystine and arginine have L and two kinds of configurational isomers of D respectively, and the biological characteristics that has of the amino acid of isomorphism type does not often have very big difference.Just have physiologically active although generally believe the natural amino acid configuration that is present in organism, also have viewpoint to think that these contain amino acid whose material, its biologically-active moiety derives from the amino acid product that degradation in vivo obtains.But studies show that, the pharmacological action of N-acetylcystein is very various, can not be fully substitutes with the L-Gelucystine of its meta-bolites, for example, the N-acetylcystein molecule has direct anti-radical action, also has the active effect of the glutathione s-transferase of enhancing (Moldeus P ﹠amp; Cotgreave IA.Meth.Enzymol.234:482-492,1994); Studies show that N-acetylcystein has chelation of metal ion, be fully by molecule itself work and with meta-bolites irrelevant (Banner W Jr.et alToxicol.Appl.Pharmacol.83:142-147,1986).N, N '-diacetyl Gelucystine has strong immunoregulation effect, and the intensity of its immunization is 1000 times of (Sarnstrand B of N-acetylcystein; Jansson AH; Matuseviciene G, et al.J Pharmacol Exp Ther.1999 Mar; 288 (3): 1174-84).N is described, N '-diacetyl Gelucystine has the pharmacological mechanism of self uniqueness, and is not only that prodrug as N-acetylcystein plays a role.In addition, N is in N '-diacetyl Gelucystine two arginic acid salt molecules, different arginine configurations had both influenced N, the physicochemical property of N '-diacetyl Gelucystine two arginic acid salt molecules influence its biological activity again, comprise pharmacological mechanism and internal metabolism kinetic parameter.Therefore, N, the different configurational isomers of N '-diacetyl Gelucystine two arginic acid salts have influences its different qualities as medicinal application.
At a part N, contain two molecule Gelucystines and two molecule arginine in N '-diacetyl Gelucystine two arginic acid salt molecules, so N, N '-diacetyl Gelucystine two arginic acid salts are to have multiple configurational isomer (N, N '-diacetyl-L-Gelucystine-L, L-two arginic acid salts, N, N '-diacetyl-D-Gelucystine-L, L-two arginic acid salts, N, N '-diacetyl-i-Gelucystine-L, L-two arginic acid salts, N, N '-diacetyl-L-Gelucystine-D, D-two arginic acid salts, N, N '-diacetyl-D-Gelucystine-D, D-two arginic acid salts, N, N '-diacetyl-i-Gelucystine-D, D-two arginic acid salts, N, N '-diacetyl-L-Gelucystine-L, D-two arginic acid salts, N, N '-diacetyl-D-Gelucystine-L, D-two arginic acid salts, N, N '-diacetyl-i-Gelucystine-L, D-two arginic acid salts, the i-Gelucystine is represented the meso Gelucystine).Usually different configurational isomers and mesomeride thereof and racemic modification have difference aspect varying degree physics chemical property and the drug effect in some occasion, particularly chiral drug be by with intravital certain macromole in the inherent binding site produce induced-fit, thereby suppress (or exciting) this macromolecular physiologically active, reach the purpose of curing the disease.The stereospecificity of biochemical reaction requires medicine to have higher optical purity, and in the process of exploitation chiral molecules medicine, preferred configuration and application optimum configuration patent medicine are important link.Therefore we are to N, and sufficient research has been carried out in the physics that various configuration had, chemical property and the pharmacological action of N '-diacetyl Gelucystine arginic acid salt, thereby has finished the present invention.
Summary of the invention
One of purpose of the present invention is the never N of isomorphism type, optimize the configuration that is suitable for medicinal application in N '-diacetyl Gelucystine two arginic acid salts, the compound of these particular configuration has immunoregulatory activity, can treat immunodeficiency diseases, autoimmune disorder, has anti-infective, antitumor action, the compound that can be used to prepare anti-infective, the antitumor drug, particularly these particular configuration that are associated with this pharmacological action has the effect for the treatment of hepatopathy liver injury, liver failure.
Another object of the present invention provides pharmaceutical composition, said composition contains preferred N, the particular configuration compound of N '-diacetyl Gelucystine two arginic acid salts is an active constituents of medicine, this pharmaceutical composition has immunoregulatory activity, can treat immunodeficiency diseases, autoimmune disorder, have anti-infective, antitumor action, can be used to prepare be associated with this pharmacological action anti-infective, the medicine of antitumor drug, has the liver injury of treatment hepatopathy especially, the effect of liver failure, contain the advantage that not only has significant pharmacological effect and pharmacokinetics in the medicine of said composition, also help and reduce production costs.
The present invention further provides the above-mentioned preferred configuration that contains as the pharmaceutical composition of active constituents of medicine and the various preparations of different way of administration, these preparations are added with or do not add medicinal carrier.
Another object of the present invention also is to provide the preparation technology of the compound of these preferred configuration.
The present invention is N more preferably, N '-diacetyl Gelucystine two arginic acid salt configurational isomers are N, N '-diacetyl-L-Gelucystine-L, L-arginic acid salt (configuration 1), N, N '-diacetyl-i-Gelucystine-L, L-arginic acid salt (configuration 3), N, N '-diacetyl-L-Gelucystine-L, D-arginic acid salt (configuration 7), N, N '-diacetyl-i-Gelucystine-L, D-arginic acid salt (configuration 9).The structure of these steric configuration compounds is as follows:
Configuration 1, N, N '-diacetyl-L-Gelucystine-L, L-arginic acid salt
Configuration 3, N, N '-diacetyl-i-Gelucystine-L, L-arginic acid salt
Configuration 7, N, N '-diacetyl-L-Gelucystine-L, D-arginic acid salt
Configuration 9, N, N '-diacetyl-i-Gelucystine-L, D-arginic acid salt
The further preferred N of the present invention, N '-diacetyl Gelucystine two arginic acid salt configurational isomers are N, N '-diacetyl-L-Gelucystine-L, L-arginic acid salt (configuration 1), N, N '-diacetyl-i-Gelucystine-L, L-arginic acid salt (configuration 3).
The most preferred N of the present invention, N '-diacetyl Gelucystine two arginic acid salt configurational isomers are N, N '-diacetyl-L-Gelucystine-L, L-arginic acid salt (configuration 1).
The invention provides a kind of N of containing, the pharmaceutical composition of N '-diacetyl Gelucystine two arginic acid salt configurational isomers, said composition contains single or blended N, N '-diacetyl-L-Gelucystine-L, L-arginic acid salt (configuration 1), N, N '-diacetyl-i-Gelucystine-L, L-arginic acid salt (configuration 3), N, N '-diacetyl-L-Gelucystine-L, D-arginic acid salt (configuration 7), N, N '-diacetyl-i-Gelucystine-L, D-arginic acid salt (configuration 9) is an active constituents of medicine.
The present invention further provides a kind of N of containing, the pharmaceutical composition of N '-diacetyl Gelucystine two arginic acid salt configurational isomers, that said composition contains is single-or blended N, N '-diacetyl-L-Gelucystine-L, L-arginic acid salt (configuration 1), N, N '-diacetyl-i-Gelucystine-L, L-arginic acid salt (configuration 3) is an active constituents of medicine.
The present invention also provides a kind of N of containing, the pharmaceutical composition of N '-diacetyl Gelucystine two arginic acid salt configurational isomers, and said composition has the N of containing, N '-diacetyl-L-Gelucystine-L, L-arginic acid salt (configuration 1) is an active constituents of medicine.
Above-mentionedly contain preferred configuration can be applied to this treatment of needs by oral, rectum or administered parenterally mode as the pharmaceutical composition of active constituents of medicine patient.Be used for when oral, can be made into conventional solid preparation such as tablet, capsule, pulvis, granule etc., make liquid preparation such as syrup, oral liquid etc.; When being used for administered parenterally, can be made into solution, powder pin, water or the oiliness suspension agent etc. of injection.Preferred form is tablet, capsule, injection liquid and injection powder pin.These preparations are added with or do not add medicinal carrier, can be by the production method preparation of pharmaceutical field routine, these methods comprise activeconstituents and one or more carrier or mixed with excipients, the method of general preparation is, closely mix activeconstituents is all even with liquid vehicle or levigated solid carrier, as required can be with the product drying and moulding.
Preparation of the present invention comprises the vehicle that meets the requirements, and these vehicle comprise tamanori, thinner, disintegrating agent, sanitas, dispersion agent, glidant and lubricant.Contain an amount of tamanori, about 5-10% disintegrating agent, about 2-5% glidant, about 1% lubricant as required.
Tablet can by active compound and one or more carrier or vehicle compresses together or mold pressing forms, the general every activeconstituents that contains the 100-500mg that has an appointment of tablet.Can carry out dressing to tablet as required, and can be mixed with the formulation that can make activeconstituents slowly-releasing or controlled release.Desirable vehicle is lactose, croscarmellose sodium, sodium starch glycolate, ethyl cellulose, talcum powder and Magnesium Stearate for example.
Injection liquid can contain antioxidant, damping fluid, isotonic agent etc.Water-based or nonaqueous suspension can contain suspension agent and thickening material.
Above-mentioned preparation can contain the medicament of commonly used other in this area, and for example, oral preparations can contain seasonings.
The mole number that medicinal compositions of the present invention contains activeconstituents accounts for the 0.1%-99.5% of whole composition mole number.
N, N '-diacetyl Gelucystine preparation has several method: method one is to be starting raw material with the N-acetylcystein, by hydrogen peroxide, again through the ion exchange resin purifying, generates N, N '-diacetyl Gelucystine under alkaline condition; Method two is to be starting raw material with the Gelucystine, uses the acetic anhydride acetylize under alkaline condition, gets N, N '-diacetyl Gelucystine.
N, N '-diacetyl Gelucystine and the salifiable way of arginine comprise the N with purifying, N '-diacetyl Gelucystine, an amount of arginine are dispersed or dissolved in respectively in solvent or the solvent mixture, both are mixed again.Solvent for use can be selected from water, alcohols, di-alcohols, ketone, amides, sulfoxide class or other polar solvents or solvent mixture.The salt that generated is through concentrating under reduced pressure, crystallization, use solvent wash, final drying finished product again.At this, in order to remove N, N '-diacetyl Gelucystine impurity can be with N, and N '-diacetyl Gelucystine solution is by Zeo-karb, water wash-out again; N, the solvent of N '-appropriate to the occasion employing of diacetyl Gelucystine two arginic acid salt crystallizations is alcohols or alcohol ketone mixed solvent, also can add suitable quantity of water in solvent.
Can also adopt method three is starting raw material by N-acetylcystein and arginine, by the directly synthetic N of hydrogen peroxide oxidation, N '-diacetyl Gelucystine two arginic acid salts.
By selecting L-Gelucystine, D-Gelucystine or mesomeric Gelucystine is starting raw material, utilizes method two to synthesize N, and N '-diacetyl Gelucystine can obtain the not N of isomorphism type, N '-diacetyl Gelucystine.Be salt forming agent with L-arginine, D-arginine single or that be mixed in proportion again, finally synthetic N, the different configurational isomers of N '-diacetyl Gelucystine two arginic acid salts.Also can select L-halfcystine or D-halfcystine, with L-arginine or D-arginine be starting raw material, utilize method one or two synthetic N, the different configurational isomers of N '-diacetyl Gelucystine two arginic acid salts.
To the synthetic N of institute, the N '-multiple configurational isomer of diacetyl Gelucystine two arginic acid salts, the easy crystallinity of assay determination, stability, water absorbability, and carry out the research of aspects such as drug metabolism, pharmacodynamics.The effect of these aspects to selecting suitable medicinal configuration to have particularly important.
The common trait that preferred configuration of compound has is to have at least one L configuration arginine in the molecule.The L-arginine is a kind of important intermediate metabolites of organism ornithine cycle.Replenish the L-arginine and can promote the urine circulation, reduce the content of ammonia in the blood, the treatment blood ammonia is poisoned, and rescues critically ill patient.When the human liver function was undesired, the urine circulation was obstructed, and ammonia content increases in the blood, will cause hepatic coma, and hepatic coma is the hepatitis gravis common complication.The L-arginine is to acute viral hepatitis, and chronic persistent hepatitis and liver cirrhosis also have better therapeutic effect, and can prevent and treat fatty live lesions, plays the effect of effectively protecting liver.Body ubiquity arginine NO pathway in cultured, in multiple physiological system, show important biology mediation characteristic and function, in liver physiology function and disease, can regulate the liver cell protein synthesis, have the important mechanism that mediates various liver poisoning and influence the liver cirrhosis hemodynamics variation.Arginine cellular metabolism approach in vivo and liver cell regeneration are in close relations, animal experiment shows, N, the arginic configuration of compound of at least one L configuration has significant curative effect mostly in N '-diacetyl Gelucystine two arginic acid salt molecules, with not salifiable L-arginine, N, N '-diacetyl-L-Gelucystine, and N, N '-diacetyl-L-Gelucystine-L-lysine salt is compared, alleviate liver injury and improve the curative effect of serological index more obvious, the survival rate of liver failure animal significantly improves, and L-arginine and N are described, N '-diacetyl Gelucystine salify has the synergy of notable synergistic attenuation to the treatment liver failure.Therefore, L-arginine and N, N '-diacetyl Gelucystine salify has not only solved parent compound and has been difficult to crystallization, is easy to the physical property of moisture absorption, and improved its biological activity.
Merge in the animal experiment that the lipopolysaccharides sensitized mice causes immunological liver depletion at GalN and to be surprisingly found out that, at N, in N '-diacetyl Gelucystine two arginic acid salt configurational isomers, N, N '-diacetyl-i-Gelucystine-L, the L-arginic acid salt, N, N '-diacetyl-L-Gelucystine-L, D-arginic acid salt, N, N '-diacetyl-i-Gelucystine-L, though contain mesomeric Gelucystine and D type arginine in the configurations such as D-arginic acid salt, with N, N '-diacetyl-L-Gelucystine-L, the L-arginic acid salt is the same, can obviously reduce the animal dead rate, improve serological index, have the ideal drug effect.It is not the naturally occurring amino acid of human body that this discovery has broken through the D type amino acid that it has been generally acknowledged that, do not have physiologically active routine understanding.
Therefore, the present invention is N more preferably, and N '-diacetyl Gelucystine two arginic acid salt configurational isomers are: N, N '-diacetyl-L-Gelucystine-L, L-arginic acid salt (configuration 1), N, N '-diacetyl-i-Gelucystine-L, L-arginic acid salt (configuration 3), N, N '-diacetyl-L-Gelucystine-L, D-arginic acid salt (configuration 7), N, N '-diacetyl-i-Gelucystine-L, D-arginic acid salt (configuration 9), have more excellent easy crystallinity, stability, be difficult for moisture absorption, drug effect is better.
Further finding in the pharmacokinetic studies, at N, in N '-diacetyl Gelucystine two arginic acid salt configurational isomers, N, N '-diacetyl-L-Gelucystine-L, L-arginic acid salt (configuration 1), N, N '-diacetyl-i-Gelucystine-L, L-arginic acid salt (configuration 3) mainly is distributed in spleen (immune organ), shows that the compound of these two configurations has significant immune organ target, helps bringing into play immunoregulation effect.
So further preferred N of the present invention, N '-diacetyl-L-Gelucystine-L, L-arginic acid salt (configuration 1), N, N '-diacetyl-i-Gelucystine-L, L-arginic acid salt (configuration 3), they are difficult for moisture absorption except having good easy crystallinity, stability, also have strong immune organ target, help drug effect and bring into play better.
Preferred these N of the present invention, N '-diacetyl Gelucystine two arginic acid salt configurational isomers not only have optimum physicochemical property and drug effect and drug metabolism characteristic, the toxicological test proof has suitable security, for the liver failure that lacks good medicine at present provides new effective treatment approach, also be suitable for preparing medicine with immunoregulatory activity, be used for anti-infective, the antitumor drug that are associated with this pharmacological action, can treat immunodeficiency diseases, autoimmune disorder.
Embodiment
The following examples can make those skilled in the art more fully understand the present invention, but do not limit the present invention in any way.
Embodiment 1 N, N '-diacetyl-L-Gelucystine-L, L-two arginic acid salts synthetic
Get potassium hydroxide 672g earlier and drop in the 5L reaction flask, add water 2400ml and be dissolved into high alkali liquid, begin gradation when being cooled to room temperature (20～30 ℃) and add L-Gelucystine 360g stirring and dissolving, be cooled to below 0 ℃.The cooling of external application ice-water bath slowly drips diacetyl oxide down, and temperature is no more than 5 ℃ in the control.Finish, continued insulated and stirred 30 minutes, placed 3 hours.Drip concentrated hydrochloric acid and regulate pH to 2～3.Get the aqueous solution of acetyl Gelucystine.
The aqueous solution underpressure distillation of acetyl Gelucystine (outer temperature 80 ℃) is concentrated into thickly, divides to extract for 3 times to filter with acetone-water (90: 10) and removes most of inorganic salt.Acetone is removed in the extracting solution underpressure distillation, is concentrated into thickness, and residue is dissolved in water (adding water 1500mL approximately).Adding 732 resin cation (R.C.)s (about 200mg) stirred 2～3 hours.
Remove by filter resin, filtrate is stirred down and is added L-arginine 523g in batches, regulates pH to 7.Continue to stir 30 minutes, reaction solution is evaporated to thick, adds the 2L alcohol heating reflux, and cooling crystallization is placed in dissolving.After separating out crystal, filter, use the small amount of ethanol washing leaching cake, filter is done, and drying under reduced pressure (40 ℃ ,-0.09MPa) 24 hours.Get N, N '-diacetyl-L-Gelucystine-L, L-two arginic acid salt crude product 909g, yield 90%.
Get above-mentioned crude product 500g, add 90% ethanol 4000ml (1: 8), heating for dissolving adds 30 minutes filtered while hot of gac 25g reflux decolour.Filtrate is put to room temperature, and refrigeration crystallization 24 hours is (below 5 ℃.Filter, and drying under reduced pressure (40 ℃ ,-0.09MPa) 24 hours, promptly get N, N '-diacetyl-L-Gelucystine-L, L-two arginic acid salt elaboration 450g, yield is 90%.This product is a white crystalline powder, and this product is easily molten in water, methyl alcohol, and slightly soluble in ethanol, fusing point are 203～208 ℃ (decomposition); Specific optical rotation is [α]
D 20-61.0～-64.0.
The results of elemental analyses of highly finished product sees the following form:
Table 1.N, N '-diacetyl-L-Gelucystine-L, the results of elemental analyses of L-two arginic acid salt highly finished product
| Test elements | C | H | N | |
| Calculated value (%) | 39.28 | 6.59 | 20.82 | |
| Measured value (%) | For the first time | 39.12 | 6.69 | 20.63 |
| For the second time | 39.18 | 6.72 | 20.69 | |
| Mean value | 39.15 | 6.71 | 20.66 |
Measurement result shows that C, the H of the two acyl Gelucystine arginic acid salt highly finished product of gained and the difference of N content measured value and theoretical value are all less than 0.3%.Results of elemental analyses shows, 1 molecule N, N '-diacetyl Gelucystine and 2 molecule arginine salifies.
By oxygen flask combustion method (two appendix VII of Chinese Pharmacopoeia version in 2000 C), adopt BaCl
2The S content of titration measuring highly finished product.S assay value is 9.33%, with the difference of theoretical value (9.53%) less than 0.3%.The S assay is the result also show, 1 molecule diacetyl Gelucystine and 2 molecule arginine salifies.
The quasi-molecular ion peak [M+H] that high resolution MS (ESI (+)) measures
+The quality at peak is 175.1191 (proofreading and correct with internal standard substance m/z206.1405, in 10.0ppm permissible error scope), and molecular formula (M) is C
6H
14N
4O
2, this composition is an arginine; The quasi-molecular ion peak [M-H] that high resolution MS (ESI (-)) measures
-The high resolution quality measurement at peak is 323.0365 (proofreading and correct with internal standard substance m/z 250.1080, in 10.0ppm permissible error scope), and molecular formula (M) is C
10H
16N
2O
6S
2, be N, N '-diacetyl Gelucystine.
Embodiment 2 N, N '-diacetyl-D-Gelucystine-L, L-two arginic acid salts synthetic
Get potassium hydroxide 670g earlier and drop in the 5L reaction flask, add water 2400ml and be dissolved into high alkali liquid, begin gradation when being cooled to room temperature and add D-Gelucystine 360g stirring and dissolving, be cooled to below 0 ℃ to (20～30 ℃).The cooling of external application ice-water bath slowly drips diacetyl oxide down, and temperature is no more than 5 ℃ in the control.Finish, continued insulated and stirred 30 minutes, placed 3 hours.Drip concentrated hydrochloric acid and regulate pH to 2～3.Get the aqueous solution of acetyl Gelucystine.
The aqueous solution underpressure distillation of acetyl Gelucystine (outer temperature 80 ℃) is concentrated into thickly, divides to extract for 3 times to filter with acetone-water (90: 10) and removes most of inorganic salt.Acetone is removed in the extracting solution underpressure distillation, is concentrated into thickness, and residue is dissolved in water (adding water 1500mL approximately).Adding 732 resin cation (R.C.)s (about 200mg) stirred 2～3 hours.
Remove by filter resin, filtrate is stirred down and is added L-arginine 523g in batches, regulates pH to 7.Continue to stir 30 minutes, reaction solution is evaporated to thick, adds the 2L alcohol heating reflux, and cooling crystallization is placed in dissolving.After separating out crystal, filter, use the small amount of ethanol washing leaching cake, filter is done, drying under reduced pressure (40 ℃ ,-0.09MPa) 24 hours.Get N, N '-diacetyl-D-Gelucystine-L, L-two arginic acid salt crude product 856g, yield 85%.
Get above-mentioned crude product 500g, add 85% ethanol 4000ml (1: 8), heating for dissolving adds gac 30g, reflux decolour 30 minutes, filtered while hot.Filtrate is put to room temperature, and refrigeration crystallization 24 hours is (below 5 ℃.Filter, and drying under reduced pressure (40 ℃ ,-0.09MPa) 24 hours, promptly get N, N '-diacetyl-D-Gelucystine-L, L-two arginic acid salt elaboration 400g, yield is 80%.The easy moisture absorption of this product crystallization, easily molten in water, methyl alcohol, slightly soluble in ethanol, fusing point are 201～206 (decomposition) ℃; Specific optical rotation is [α]
D 20+ 115.4～+ 120.0 °.Ultimate analysis calculated value: C
22HL
44N
10S
2O
10, C:39.28 H:6.59 N:20.82; Measured value: C:39.17 H:6.70 N:20.68.S assay value is 9.34%, theoretical value (9.53%).
Embodiment 3 N, N '-diacetyl-L-Gelucystine-D, D-two arginic acid salts synthetic
Get potassium hydroxide 675g earlier and drop in the 5L reaction flask, add water 2400ml and be dissolved into high alkali liquid, begin gradation when being cooled to room temperature and add L-Gelucystine 360g stirring and dissolving, be cooled to below 0 ℃ to (20～30 ℃).The cooling of external application ice-water bath slowly drips diacetyl oxide down, and temperature is no more than 5 ℃ in the control.Finish, continued insulated and stirred 30 minutes, placed 3 hours.Drip concentrated hydrochloric acid and regulate pH to 2～3.Get the aqueous solution of acetyl Gelucystine.
The aqueous solution underpressure distillation of acetyl Gelucystine (outer temperature 80 ℃) is concentrated into thickly, divides to extract for 3 times to filter with acetone-water (90: 10) and removes most of inorganic salt.Acetone is removed in the extracting solution underpressure distillation, is concentrated into thickness, and residue is dissolved in water (adding water 1500mL approximately).Adding 732 resin cation (R.C.)s (about 200mg) stirred 2～3 hours.
Remove by filter resin, filtrate is stirred down and is added D-arginine 523g in batches, regulates pH to 7.Continue to stir 30 minutes, reaction solution is evaporated to thick, adds the 2L alcohol heating reflux, and cooling crystallization is placed in dissolving.After separating out crystal, filter, use the small amount of ethanol washing leaching cake, filter is done, and drying under reduced pressure (40 ℃ ,-0.09MPa) 24 hours.Get N, N '-diacetyl-L-Gelucystine-D, D-two arginic acid salt crude product 900g, yield 90%.
Get above-mentioned crude product 500g, add 90% ethanol 4000ml (1: 8), heating for dissolving adds gac 25g reflux decolour 30 minutes, filtered while hot.Filtrate is put to room temperature, and refrigeration crystallization 24 hours is (below 5 ℃.Filter, and drying under reduced pressure (40 ℃ ,-0.09MPa) 24 hours, promptly get N, N '-diacetyl-L-Gelucystine-L, L-two arginic acid salt elaboration 435g, yield is 87%.This product is that crystallization is slow, easily moisture absorption, and easily molten in water, methyl alcohol, slightly soluble in ethanol, fusing point are 201～205 ℃ (decomposition); Specific optical rotation is [α]
D 20-83.7～87.2 °.Ultimate analysis calculated value: C
22H
44N
10S
2O
10, C:39.28 H:6.59 N:20.82; Measured value: C:39.20 H:6.67 N:20.70.S assay value is 9.36%, theoretical value (9.53%).
Embodiment 4 N, N '-diacetyl-i-Gelucystine-L, L-two arginic acid salts synthetic
Get potassium hydroxide 670g earlier and drop in the 5L reaction flask, add water 2400ml and be dissolved into high alkali liquid, begin gradation when being cooled to room temperature and add i-Gelucystine 360g stirring and dissolving, be cooled to below 0 ℃ to (20～30 ℃).The cooling of external application ice-water bath slowly drips diacetyl oxide down, and temperature is no more than 5 ℃ in the control.Finish, continued insulated and stirred 30 minutes, placed 3 hours.Drip concentrated hydrochloric acid and regulate pH to 2～3.Get the aqueous solution of acetyl Gelucystine.
The aqueous solution underpressure distillation of acetyl Gelucystine (outer temperature 80 ℃) is concentrated into thickly, divides to extract for 3 times to filter with acetone-water (90: 10) and removes most of inorganic salt.Acetone is removed in the extracting solution underpressure distillation, is concentrated into thickness, and residue is dissolved in water (adding water 1500mL approximately).Adding 732 resin cation (R.C.)s (about 200mg) stirred 2～3 hours.
Remove by filter resin, filtrate is stirred down and is added L-arginine 520g in batches, regulates pH to 7.Continue to stir 30 minutes, reaction solution is evaporated to thick, adds the 2L alcohol heating reflux, and cooling crystallization is placed in dissolving.After separating out crystal, filter, use the small amount of ethanol washing leaching cake, filter is done, drying under reduced pressure (40 ℃-0.09MPa) 24 hours, get N, N '-diacetyl-i-Gelucystine-L, L-two arginic acid salt crude product 911g, yield 90%.
Get above-mentioned crude product 500g, add 90% ethanol 4000ml (1: 8), heating for dissolving adds gac 25g reflux decolour 30 minutes, filtered while hot.Filtrate is put to room temperature, and refrigeration crystallization 24 hours is (below 5 ℃.Filter, and drying under reduced pressure (40 ℃ ,-0.09MPa) 24 hours, promptly get N, N '-diacetyl-i-Gelucystine-L, L-two arginic acid salt elaboration 452g, yield is 90%.This product is a white crystalline powder, is difficult for moisture absorption, and is easily molten in water, methyl alcohol, slightly soluble in ethanol; Fusing point is 160～162 ℃; Specific optical rotation is [α]
D 20+ 26.9～+ 27.9 °.Ultimate analysis calculated value: C
22H
44N
10S
2O
10, C:39.28 H:6.59 N:20.82; Measured value: C:39.22 H:6.69 N:20.70.S assay value is 9.35%, theoretical value (9.53%).
Embodiment 5 N, N '-diacetyl-L-Gelucystine-L, D-two arginic acid salts synthetic
Restrain N-acetyl-L-cysteines and 150 gram L-arginine and 150 gram D-arginine with 300 and be dissolved in the 1000ml deionized water, slowly add hydrogen peroxide 80ml while stirring, keep temperature to be lower than below 25 ℃ continuously stirring 5 hours.Add 1000ml95% ethanol in solution, it is complete to continue to be stirred to crystallization.With ethanol 50ml washing, drying under reduced pressure (40 ℃-0.09MPa) 24 hours, get N, N '-diacetyl-L-Gelucystine-L, D-two arginic acid salts 500 grams, yield 82%.This product is a white crystalline powder, is difficult for moisture absorption, and is easily molten in water, methyl alcohol, slightly soluble in ethanol; Fusing point is 196～199 ℃ (decomposition); Specific optical rotation is [α]
D 20-58.0～-61.2 °.Ultimate analysis calculated value: C
22H
44N
10S
2O
10, C:39.28 H:6.59 N:20.82; Measured value: C:39.22 H:6.69 N:20.70.S assay value is 9.35%, theoretical value (9.53%).
Embodiment 6 N, N '-diacetyl-i-Gelucystine-L, D-two arginic acid salts synthetic
200 gram N-acetyl-L-cysteines, 200 gram N acetyl-D-halfcystines and 200 gram L-arginine, 200 gram D-arginine are dissolved in the 1000ml deionized water, slowly add hydrogen peroxide 80ml while stirring, keep temperature to be lower than below 25 ℃ continuously stirring 5 hours.Add 1000ml95% ethanol in solution, it is complete to continue to be stirred to crystallization.With ethanol 50ml washing, drying under reduced pressure (40 ℃-0.09MPa) 24 hours, get N, N '-diacetyl-i-Gelucystine-L, D-two arginic acid salts 650 grams, yield 80%.This product is a white crystalline powder, is difficult for moisture absorption, and is easily molten in water, methyl alcohol, slightly soluble in ethanol; Fusing point is 156～158 ℃ (decomposition); Specific optical rotation is [α]
D 20+ 16.5～+ 19.1 °.Ultimate analysis calculated value: C
22H
44N
10S
2O
10, C:39.28 H:6.59 N:20.82; Measured value: C:39.22 H:6.65 N:20.74.S assay value is 9.36%, theoretical value (9.53%).
Embodiment 7 N, N '-diacetyl Gelucystine two arginic acid salt configurational isomer physicochemical property
| Configurational isomer | Fusing point (℃) | Specific optical rotation [α] D 20(°) | |
| 1 | N, N '-diacetyl-L-Gelucystine-L, L-two arginic acid salts | 203～208 (decomposition) | -61.0～-64.0 |
| 2 | N, N '-diacetyl-D-Gelucystine-L, L-two arginic acid salts | 201～206 (decomposition) | +115.4～+120.0 |
| 3 | N, N '-diacetyl-i-Gelucystine-L, L-two arginic acid salts | 160～162 (decomposition) | +26.9～+27.9 |
| 4 | N, N '-diacetyl-L-Gelucystine-D, D-two arginic acid salts | 201～205 (decomposition) | -83.7～87.2 |
| 5 | N, N '-diacetyl-D Gelucystine-D, D-two arginic acid salts | 204～208 (decomposition) | +145.2～+150.2 |
| 6 | N, N '-diacetyl-i-Gelucystine-D, D-two arginic acid salts | 154～159 (decomposition) | -40.2～-42.8 |
| 7 | N, N '-diacetyl-L-Gelucystine-L, D-two arginic acid salts | 196～199 (decomposition) | -58.0～-61.2 |
| 8 | N, N '-diacetyl-D-Gelucystine-L, D-two arginic acid salts | 178～181 (decomposition) | +124.1～+127.0 |
| 9 | N, N '-diacetyl-i-Gelucystine-L, D-two arginic acid salts | 156～158 (decomposition) | +16.5～+19.1 |
Embodiment 8 N, the curative effect method summary of N '-diacetyl Gelucystine two arginic acid salt configurational isomers treatment mouse immune liver failure: to mouse peritoneal injection (ip) GalN (GalN) 1000mg/kg, while ip lipopolysaccharides (LPS) 10 μ g/kg, the administration group is in injecting the forward and backward 1h of GalN/LPS ip.N respectively, N '-diacetyl Gelucystine two arginic acid salt configurational isomer 200mg/kg, measure the serum transaminase level behind the 6h, get anticoagulated whole blood and measure CD4+, CD8+, TH1, TH2, get hepatic tissue and do the pathology observation, and observe the 24h death condition.
The summary of results: 1,3,7,9 type configurational isomers can suppress GalN/LPS and attack the mice serum ALT, the AST vigor that cause behind the 6h and increase, and obviously increase CD8+, TH1, TH2 lymphocyte, and the hepatic tissue infringement is alleviated, and improve the mouse survival rate.Illustrate that 1,3,7,9 type configurational isomers cause the mouse immune liver failure to Gal/LPS obvious prevention and therapeutic action are arranged.Especially best with 1 type configurational isomer curative effect.
[reagent preparation] gets N, N '-diacetyl Gelucystine two arginic acid salt configurational isomers, be mixed with the solution that every 1ml contains 20mg respectively with 0.9% sodium chloride solution, pH is 6.5 ~ 8.0, filter with the 4# sintered filter funnel, can in the 10ml ampoule, sealing by fusing, sterilized 45 minutes for 105 ℃, face with preceding with physiological saline dilution or directly application.
10 every group of [experimental animal] BalB/C ♂ mouse.Sex, body weight: ♂ mouse 16～18g.All animals of duration of test except that the back fasting 20h (freely drinking water) that poisons, equal ad lib of all the other times and drinking-water.
[test key step] gets 150 of BALB/C ♂ mouse, divides 15 groups at random, 10 every group.Except that negative control group, all animals are ip 12%GalN solution (containing lipopolysaccharides 10 μ g/kg) 1000mg/kg simultaneously, the 1h and the back 1h ip N respectively that poisons before poisoning are organized in treatment, N '-diacetyl Gelucystine two arginic acid salt configurational isomer 200mg/kg, 1 group of ip hepatocyte growth-promoting factors of positive control (pHGF) 60mg/kg (document dosage), 2 groups of ip N of positive control, N '-diacetyl-L-Gelucystine 100mg/kg, 3 groups of ip L-of positive control arginine 100mg/kg, 4 groups of ip N of positive control, N '-diacetyl-L-Gelucystine-L-lysine salt 200mg/kg.Model and normal control group ip co-content physiological saline, respectively group is observed the survival condition of 24h, and after injecting LPS/Gal 6h, puts to death part zoometry Serum ALT and AST, taking heparin anticoagulated whole blood cells were tested by flow cytometry CD4+, CD8+, TH1, TH2; The left lobe of liver system pathological section of getting is done histological examination.
[experimental control]
Normal control group ip co-content blank solvent is to get rid of solvent effect.
The frequency injection of model control group ip co-content blank solvent in the hope of being equal to.
The positive controls positive control adopts the clinical hepatocyte growth-promoting factors (pHGF) that is used for the treatment of hepatitis gravis at present for 1 group, and pharmaceutical factory, Yangjiang, Guangdong produces (94) and defends lot number the accurate word x-67 of medicine number: 021013, and the 20mg/ bottle,, and according to document dosage.Show according to the pHGF pharmacodynamic study, can promote liver failure rat DNA synthetic, suppress TNF, improve blood circulation and reduce mortality ratio, therefore be subjected to the purposes basically identical of reagent thing and positive control medicine.Positive control adopts the synthetic N of institute of Jiangsu Zhengda Tianqing Drug Industry Co., Ltd for 2 groups, N '-diacetyl-L-Gelucystine 100mg/kg, 3 groups of ip of positive control adopt L-arginine (Sigma company) 100mg/kg, positive control adopts the synthetic N of institute of Jiangsu Zhengda Tianqing Drug Industry Co., Ltd, N '-diacetyl-L-Gelucystine-L-lysine salt 200mg/kg for 4 groups.
[statistical method] test-results is asked mean
With standard deviation (s), remove mortality ratio test of significance x
2Check, other test of significance non-paired t test, histopathology adopts the check of grade preface value method.P≤0.05 or P≤0.01 thinks to have statistical significance.
[experimental result]
1, to GalN/LPS cause the liver failure mouse death rate influence ip in mice GalN/LPS 6h after, diet, activity obviously reduce, dispirited, drowsiness, perpendicular hair appears in most mouse, symptom aggravation behind the 8h, stupor, hemorrhage and begin death, the symptom of configuration 1,3,7,9 treatment groups (200mg/kg) is obviously light than model control group, and the curative effect that reduces mortality ratio is better than positive control drug, it is very fast that surviving animals is recovered, and diet, activity obviously increase behind the 24h.The results are shown in Table one.Test-results shows that configuration 1,3,7,9 has significant curative effect.
Table one, to LPS/Gal cause the liver failure mouse death rate influence (
)
| Test group | Number of animals | Dosage (mg/kg) | Mortality ratio (%) |
| 4 groups of configurations of 3 groups of positive controls of 2 groups of positive controls of 1 group of positive control of model control group Normal group positive control, 1 treatment group configuration, 2 treatment group configurations, 3 treatment group configurations, 4 treatment group configurations, 5 treatment group configurations, 6 treatment group configurations, 7 treatment group configurations, 8 treatment group configurations, 9 treatment groups | 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 | 60 100 100 200 200 200 200 200 200 200 200 200 200 | 80 0 50 ** 70 * 80 * 70 ** 30 ** 60 ** 40 ** 60 ** 60 ** 60 ** 30 ** 70 ** 40 * |
*P＜0.05, compare with model control group * * P＜0.01.
2, GalN/LPS is caused the influence of liver failure mice serum ALT and AST vigor
The serum transaminase level increases slightly behind the ip in mice GalN/LPS 5h, and the vigor that configuration 1,3,7,9 treatment groups can obviously suppress transaminase improves serological index, sees the following form
Table two, to LPS/Gal cause liver failure mice serum ALT and AST vigor influence (
)
| Test group | Number of animals | Dosage (mg/kg) | ALT(IU/L) | AST(IU/L) |
| 4 groups of configurations of 3 groups of positive controls of 2 groups of positive controls of 1 group of positive control of model control group Normal group positive control, 1 treatment group configuration, 2 treatment group configurations, 3 treatment group configurations, 4 treatment groups | 10 10 10 10 10 10 10 10 10 10 | 60 100 100 200 200 200 200 200 | 98.5±25.3 42.5±10.6 ** 63.1±24.7 ** 81.9±25.6 ** 93.2±23.4 ** 80.2±23.7 ** 63.4±12.3 ** 84.6±15.4 ** 67.7±17.1 ** 73.9±11.3 ** | 203.2±56.8 83.6±15.6 ** 139.1±33.2 ** 176.2±30.3 ** 179.3±34.5 ** 173.3±31.6 ** 142.0±34.2 ** 172.0±44.1 ** 157.2±36.8 ** 158.0±30.8 ** |
| Configuration 5 treatment group configurations 6 treatment group configurations 7 treatment group configurations 8 treatment group configurations 9 treatment groups | 10 10 10 10 10 | 200 200 200 200 200 | 94.6±25.2 ** 75.8±18.4 ** 64.7±13.2 ** 91.6±26.3 ** 70.6±17.6 ** | 192.0±44.7 ** 155.2±31.6 ** 142.0±28.3 ** 188.0±42.5 ** 140.2±30.5 ** |
*P＜0.05, compare with model control group * * P＜0.01.
3, to the influence of the depleted mouse T cell proliferation and differentiation of immunological liver
GalN/LPS injection back 6h, mouse CD4+, CD8+T lymphocyte obviously reduce, and TH1, TH2 lymphocyte slightly increase.Obviously increase the CD8+ lymphocyte with model control group comparison 1,3,7,9 type configuration treatment groups, and raise TH1, TH2 cytoactive, the depleted mouse state of an illness of immunological liver is alleviated, mortality ratio descends.Test-results sees the following form:
Table three, to the influence of the depleted mouse T cell proliferation and differentiation of immunological liver (
)
| Test group | Number of animals | Dosage (mg/kg) | CD4+(％) | CD8+(％) | TH1(％) | TH2(％) |
| 4 groups of configurations of 3 groups of positive controls of 2 groups of positive controls of 1 group of positive control of Normal group model control group positive control, 1 treatment group configuration, 2 treatment group configurations, 3 treatment group configurations, 4 treatment group configurations, 5 treatment group configurations, 6 treatment group configurations, 7 treatment group configurations, 8 treatment group configurations, 9 treatment groups | 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 | 60 100 100 200 200 200 200 200 200 200 200 200 200 | 57.0±14.7 **36.6±13.2 36.4±15.6 **42.2±16.3 **36.4±15.1 **41.4±16.2 **54.8±15.1 **38.1±10.0 *50.5±12.2 **31.4±12.3 **32.8±10.1 **40.0±11.0 **53.4±11.7 **35.6±9.4 **45.2±10.3 ** | 5.0±3.3 **2.0±1.0 3.3±1.5 **3.3±0.8 **2.9±0.4 **3.0±0.7 **12.6±4.1 **8.7±3.2 *11.9±3.1 **9.7±3.0 **7.7±2.7 **8.6±2.5 **12.1±3.0 **2.90±0.2 **11.6±3.3 ** | 0.30±0.19 ** 0.39±0.28 0.20±0.13 * 1.00±0.53 ** 0.40±0.24 ** 1.10±0.62 ** 3.51±1.47 ** 0.91±0.32 * 3.01±1.40 ** 2.91±0.82 ** 1.12±0.40 ** 2.01±1.02 ** 3.44±0.89 ** 0.96±0.10 ** 2.74±0.42 ** | 0.28±0.10 ** 0.34±0.27 0.28±0.08 * 1.28±0.08 ** 0.88±0.38 ** 1.18±0.89 ** 4.37±1.21 ** 1.40±1.08 * 4.40±2.02 ** 2.00±1.18 ** 2.00±0.09 ** 3.30±0.69 ** 4.42±1.08 ** 0.89±0.17 ** 3.30±0.69 ** |
*P＜0.05, compare with model control group * * P＜0.01.
4, GalN/LPS is caused the influence of liver failure murine liver tissue
Table four, GalN/LPS cause the number of animals of liver failure mouse liver tissue injuries at different levels
| Test group | Number of animals | Dosage (mg/kg) | 3 grades | 2 grades | 1 grade | 0 grade | Average progression |
| 3 groups of 2 groups of positive controls of 1 group of positive control of normal control group model control group positive control | 10 10 10 10 10 | 60 100 100 | 0 6 1 3 3 | 0 3 7 6 7 | 0 1 2 1 0 | 10 0 0 0 0 | 0 2.5 1.9 2.2 2.3 |
| 4 groups of configurations of positive control, 1 treatment group configuration, 2 treatment group configurations, 3 treatment group configurations, 4 treatment group configurations, 5 treatment group configurations, 6 treatment group configurations, 7 treatment group configurations, 8 treatment group configurations, 9 treatment groups | 10 10 10 10 10 10 10 10 10 10 | 200 200 200 200 200 200 200 200 200 200 | 3 1 2 1 3 3 2 1 2 1 | 6 1 2 2 4 3 3 3 5 4 | 1 8 6 7 3 4 5 6 3 5 | 0 0 0 0 0 0 0 0 0 0 | 2.2 1.3 1.6 1.4 2.1 1.9 1.7 1.5 1.9 1.6 |
Number of animals/every treated animal number of average progression=damage rank * appropriate level
Embodiment 9 N, N '-diacetyl Gelucystine two arginic acid salt configurational isomers are in the intravital tissue distribution of mouse
Get 12 of Kunming kind small white mouses, be divided into 3 groups at random, every group 4, tail vein injection gives the N of 80mg/kg respectively, N '-diacetyl-L-Gelucystine-L, L-arginic acid salt (configuration 1), N, N '-diacetyl-i-Gelucystine-L, L-arginic acid salt (configuration 3), N, N '-diacetyl-L-Gelucystine-L, D-arginic acid salt (configuration 7), N, N '-diacetyl-i-Gelucystine-L, D-arginic acid salt (configuration 9), and after administration 5,30, and 60 minutes execution animals are cored, liver, spleen, lung, kidney, brain, muscle, intestines, stomach, testis, tissues such as uterus are also weighed, add physiological saline 2.0mL respectively and make homogenate, centrifugation is then got supernatant liquor 10 μ L and is analyzed under selected chromatographic condition, calculates contained sample concentration with marker method.
Suitable chromatographic condition: LichrospherC
18Post (4.6mm 15cm, dp5 μ m), moving phase is methyl alcohol: 0.025mol/L potassium primary phosphate (PH3.0)=10: 90 (v/v).Detect wavelength 205nm.
Test-results shows, N, N '-diacetyl-L-Gelucystine-L, L-arginic acid salt (configuration 1), N, N '-diacetyl-i-Gelucystine-L, L-arginic acid salt (configuration 3) is higher than far away at its hetero-organization at the tissue content of spleen, points out these two kinds of configuration of compound to have the spleen target, and other configuration of compound do not have this target.Test-results sees the following form:
Table five N, N '-diacetyl Gelucystine two arginic acid salt configurational isomers are put forward intravital tissue distribution (n=4) mouse
| Average tissue distribution (60min, μ g/g) | Configuration 1 | Configuration 3 | Configuration 7 | Configuration 9 |
| The heart | 0 | 0 | 0 | 0 |
| Liver | 0 | 0 | 20.9±2.8 | 41.2±3.6 |
| Spleen | 110.6±20.2 | 102.8±18.1 | 30.5±5.6 | 22.8±3.4 |
| Lung | 0 | 0 | 0 | 0 |
| Intestines | 15.0±3.5 | 25.4±3.2 | 62.3±10.6 | 68.4±8.0 |
| Kidney | 3.6±1.1 | 3.2±1.0 | 11.6±1.8 | 15.3±2.0 |
| Stomach | 0 | 0 | 0 | 0 |
| Brain | 0 | 0 | 0 | 0 |
| Muscle | 0 | 0 | 0 | 0 |
| The uterus | 0 | 0 | 0 | 0 |
| Testis | 0 | 0 | 0 | 0 |
Embodiment 10 injection liquids
Prescription:
N, N '-diacetyl-L-Gelucystine-L, L-two arginic acid salt 10g
Sodium-chlor 9g
Water for injection adds to 1000ml
Sodium-chlor added in about 800ml water for injection to stir make dissolving, add N, N '-diacetyl-L-Gelucystine-L, L-two arginic acid salts are regulated pH to 7.0, add water to full dose, be heated to about 55 ℃, add needle-use activated carbon, charcoal is taken off in insulation absorption 30 minutes, filter ball, filter membrane, embedding, sterilization, packing, promptly.Can be sub-packed in the infusion bottle of 250ml, 500ml.
Embodiment 11 injectable sterile powders
With N, N '-diacetyl-L-Gelucystine-L after L-two arginine are dissolved in water for injection, add gac and filters, and spray-dried, aseptic subpackaged, sealing gets Injectable sterile packing product.Specification can be made into 25mg, 50mg, 100mg, 500mg, 1g.
Embodiment 12 capsules
Prescription:
N, N '-diacetyl-L-Gelucystine-L, L-two arginic acid salt 150g
N, N '-diacetyl-i-Gelucystine-L, L-two arginic acid salt 150g
Pregelatinized Starch 150g
Magnesium Stearate 1.5g
Make 1000 capsules
With N, N '-diacetyl-L-Gelucystine-L, L-two arginic acid salts, N, N '-diacetyl-i-Gelucystine-L, L-two arginic acid salt separated pulverizing sieve, and add pregelatinized Starch, Magnesium Stearate, and mixing incapsulates shell, promptly.
Embodiment 13 tablets
Prescription:
N, N '-diacetyl-L-Gelucystine-L, L-two arginic acid salt 500g
Lactose 150g
Sodium starch glycolate 100g
Microcrystalline Cellulose 80g
Micropowder silica gel 10g
Magnesium Stearate 5g
85% ethanol is an amount of
Make 1000
With N, N '-diacetyl-L-Gelucystine-L, the sodium starch glycolate mixing of L-two arginic acid salts, lactose, Microcrystalline Cellulose and half amount, use 85% alcohol granulation, behind oven dry, the whole grain, add residue sodium starch glycolate, micropowder silica gel, Magnesium Stearate mixing, compressing tablet, promptly.
Embodiment 14 tablets
Prescription:
N, N '-diacetyl-L-Gelucystine-L, L-two arginic acid salt 10g
Lactose 10g
Sodium starch glycolate 10g
Microcrystalline Cellulose 8g
Micropowder silica gel 12g
Magnesium Stearate 0.6g
85% ethanol is an amount of
Make 1000
With N, N '-diacetyl-L-Gelucystine-L, the sodium starch glycolate mixing of L-two arginic acid salts, lactose, Microcrystalline Cellulose and half amount, use 85% alcohol granulation, behind oven dry, the whole grain, add residue sodium starch glycolate, micropowder silica gel, Magnesium Stearate mixing, compressing tablet, promptly.
Claims (11)
1, N, N '-diacetyl Gelucystine two arginic acid salt configurational isomers is characterized in that Gelucystine is L type or meso-form in the molecule, and arginic configuration is to contain a L type arginine at least in the molecule simultaneously, and structure is as follows:
2, the described N of claim 1, the configurational isomer of N '-diacetyl Gelucystine two arginic acid salts, structure is as follows:
3, the described N of claim 1, the configurational isomer of N '-diacetyl Gelucystine two arginic acid salts, structure is as follows:
4, a kind of pharmaceutical composition is characterized in that containing the described N of one or more claims 1, and the configurational isomer of N '-diacetyl Gelucystine two arginic acid salts is its activeconstituents.
5, the described pharmaceutical composition of claim 4 is characterized in that containing the described N of one or more claims 2, and the configurational isomer of N '-diacetyl Gelucystine two arginic acid salts is its activeconstituents.
6, the described pharmaceutical composition of claim 5 is characterized in that containing the described N of claim 3, and the configurational isomer of N '-diacetyl Gelucystine two arginic acid salts is its activeconstituents.
7, the described pharmaceutical composition of claim 4 is characterized in that also containing in the composition pharmaceutical carrier.
8, the described pharmaceutical composition of claim 7, it is tablet, capsule, injection liquid or injection powder pin.
9, the preparation method of the arbitrary compound of claim 1-3; this method comprises that with L-Gelucystine, D-Gelucystine or mesomeric Gelucystine be starting raw material; under alkaline condition, use the acetic anhydride acetylize; obtain the di-acetyl Gelucystine; add optically pure L-arginine or D-arginine again; dispersing and dissolving, concentrating under reduced pressure, crystallization, washing, final drying finished product.
10, the preparation method of the arbitrary compound of claim 1-3, this method comprises that L-halfcystine, D-halfcystine or the mesomeric halfcystine with N-acetyl is starting raw material, under alkaline condition, pass through hydrogen peroxide, purifying obtains the di-acetyl Gelucystine, add optically pure L-arginine or D-arginine again, dispersing and dissolving, concentrating under reduced pressure, crystallization, washing, final drying finished product.
11, the arbitrary compound of claim 1-3 is being used to prepare the medicine with immunoregulatory activity, the application in the medicine of the medicine of antitumor drug, treatment immunodeficiency diseases, autoimmune disorder, treatment infectious diseases or the liver injury of treatment hepatopathy, liver failure.
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