WO2017173609A1 - 一种抗红斑狼疮联合免疫用药物及其制备方法、用途 - Google Patents

一种抗红斑狼疮联合免疫用药物及其制备方法、用途 Download PDF

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WO2017173609A1
WO2017173609A1 PCT/CN2016/078649 CN2016078649W WO2017173609A1 WO 2017173609 A1 WO2017173609 A1 WO 2017173609A1 CN 2016078649 W CN2016078649 W CN 2016078649W WO 2017173609 A1 WO2017173609 A1 WO 2017173609A1
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extract
capsule
taking
walnut
american ginseng
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PCT/CN2016/078649
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English (en)
French (fr)
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鲁伟
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北京路德凯奇商贸有限责任公司
鲁伟
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Priority to PCT/CN2016/078649 priority Critical patent/WO2017173609A1/zh
Priority to EP16897544.9A priority patent/EP3441078A4/en
Publication of WO2017173609A1 publication Critical patent/WO2017173609A1/zh

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
    • A61K9/16—Agglomerates; Granulates; Microbeadlets ; Microspheres; Pellets; Solid products obtained by spray drying, spray freeze drying, spray congealing,(multiple) emulsion solvent evaporation or extraction
    • A61K9/1682—Processes
    • A61K9/1688—Processes resulting in pure drug agglomerate optionally containing up to 5% of excipient
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate

Definitions

  • the invention belongs to the field of medicines, and more particularly relates to a medicine for combined anti-erysue combined with immunization, a preparation method thereof and use thereof.
  • Lupus Erythematosus is an autoimmune connective tissue disease.
  • Lupus erythematosus is divided into discoid lupus erythematosus (DLE), subacute cutaneous lupus erythematosus (SCLE), systemic lupus erythematosus (SLE), deep lupus erythematosus (LEP), neonatal lupus erythematosus (NLE), drug erythema Subtypes such as lupus (DIL).
  • DLE discoid lupus erythematosus
  • SCLE subacute cutaneous lupus erythematosus
  • SLE systemic lupus erythematosus
  • LEP deep lupus erythematosus
  • NLE neonatal lupus erythematosus
  • DIL drug erythema Subtypes such as lupus (DIL).
  • the etiology of lupus erythematosus is not fully understood, but the medical community generally believes that the etiology of lupus erythematosus may be related to the immune function disorder caused by various factors such as heredity, sex hormones and environment.
  • the treatment of lupus erythematosus mainly includes Western medicine western medicine therapy and Chinese medicine method.
  • Western medicine has a certain effect on the treatment of lupus erythematosus, especially in the outbreak period or acute active period of the disease.
  • the use of large doses of hormones and cyclophosphamide drugs has a positive effect on the rapid control of the disease, but the long-term use of these drugs will cause side effects.
  • the object of the present invention is to provide a bacteriostatic and bactericidal activity, enhance the body immunity, and quickly and effectively treat the disease of lupus erythematosus; further enhance the body immunity, adjust the internal environment, and restore the body.
  • the present invention provides the following technical solutions:
  • Anti-lubris lupus combined immunization drug which is derived from walnut extract, Xiang Ai Taking the extract, American ginseng extract and Dendrobium extract as raw materials; the weight ratio of the walnut extract, the fragrant extract, the American ginseng extract and the Dendrobium extract is 1: (0.5-2): (1 ⁇ 5): (1 ⁇ 10).
  • the walnut is black walnut
  • the fragrant worm is Zhongya wormwood
  • the American ginseng is semi-wild American ginseng
  • the sarcophagus is wild sarcophagus.
  • the walnut extract of the present invention is prepared by endothelium extraction of Black Walnut (Juglans nigra L), which contains walnut, flavonoids, tannins, polysaccharides, gallic acid, volatile oil, plant viscose, white Protein, minerals, cellulose, linoleic acid, glycerides, linolenic acid, glyceryl oleate, riboflavin, carotene, salicin, casuarina, thorn fruit, research found that its active ingredients
  • the purpose of the present invention can be attained by different ratios, and the walnut extract prepared by the preparation method of the present invention has the best effect.
  • Xiang Ai also known as Ai Ye, Artemisia, Ai Ai or Jia Ai
  • the fragrant extract of the present invention is preferably prepared by whole grass extraction of North American woormwood (or Artemisia absinthium), the main component of which is volatile oil, and terpene enol-4 (Terpinenol-4) in the oil.
  • American ginseng also known as American ginseng, American ginseng, and American ginseng
  • Semi-wild American ginseng is one of the American ginseng varieties of American ginseng. Its main producing areas are 25 states including Wisconsin, New York, and Kentucky. It is cultivated by placing the seeds of American ginseng in a wild-like environment.
  • the American ginseng extract of the invention is prepared by the stepwise extraction of semi-wild American ginseng, and its main components are ginsenosides, volatile oils, fatty acids in oils and fats, organic acids, amino acids, trace elements, carbohydrates, pectin, sterols, ginsenosides.
  • Saponins Ro, Rb 1 , Rb 2 , Rb 3 , Rc, Rd, Re, Rf, Rg 1 , Rg 2 , Rg 3 , Rh 1 , R AO , American ginsenoside R 1 , Gynostemma sinensis, esculin XVII and pseudo ginseng saponins are mainly F 11 , ginseng soap F 2 , F 3 , etc.
  • the volatile oils are mainly ⁇ -farnesene, sesquiterpenoids, myrrhene, ⁇ -curcumene, etc.
  • the purpose of the present invention can be attained by different ratios, and the American ginseng extract prepared by the preparation method of the present invention has the best effect.
  • Dendrobium alias for spider plant, Duran, which is Dendrobium nobile Lindl., Dendrobium candidum Wall.ex Lindl. or Dendrobium fimbriatum Hook.Var. Stem of oculatum Hook and its approximate species. Its sweet, slightly cold, Yiwei Shengjin, nourishing yin and clearing heat.
  • the Dendrobium extract of the present invention is prepared by extracting the stem of wild Dendrobium, and contains various alkaloids, polysaccharides, amino acids, mucilage, starch, phenanthrene and bibenzyl.
  • the alkaloids include Dendrobine and Dendrobium.
  • Dendrine, Dendramine, Dendroxine, 6-Hydroxy-dendroxine, etc. have found that the ratio of active ingredients can reach the purpose of the present invention. And the effect of the Dendrobium extract prepared by the preparation method of the present invention is the best.
  • the weight ratio of the walnut extract, the fragrant extract, the American ginseng extract and the extract of Dendrobium extract of the anti-lubricated erythematosus combined with the present invention is 1: (0.5-2): (1 to 5): (1 ⁇ 10), its combination synergy:
  • the weight ratio of the walnut extract, the fragrant extract, the American ginseng extract and the sarcophagus extract is preferably 1:1:5:10.
  • the formulation of the drug is a capsule.
  • the administration of the capsules is advantageous for ensuring that the anti-lubricated erythematosus combined immunization drug of the present invention can be administered accurately and conveniently, and is beneficial for control.
  • the preparation of the walnut extract capsule, the fragrant extract capsule, the American ginseng extract capsule, and the sarcophagus extract capsule include:
  • step S2 taking the raw material softened by the step S1, supercritical fluid extraction, separating the extract and the residue;
  • step S3 taking the residue obtained in step S2, pulverizing and grinding;
  • step S4 taking the extract obtained in step S2, spray-drying, adding the pulverized and ground residue obtained in step S3 during the spray drying process to obtain a dried raw material extract;
  • step S5 taking the raw material extract obtained in the formula amount of step S4, and filling it into the capsule shell to obtain a capsule.
  • the invention adopts the vacuum cold dip softening method, rapidly inhales water by the action of negative pressure, accelerates the softening of the medicine, ensures softening of the medicine with water at normal temperature, shortens the infiltration time, reduces the loss of the active ingredient, damage and the mildew of the medicine.
  • the invention adopts supercritical fluid extraction technology to extract medicinal plants, and the fluid extracted by the supercritical fluid is carbon dioxide, and the extraction temperature is 30-40 ° C, which can avoid the volatilization of volatile oil and other components in the extraction process, and can maintain the medicine to the greatest extent.
  • the biologically active ingredients of the plant are used; at the same time, because the organic solvent is not applicable in the process, no solvent remains, which ensures the natural activity of the raw material, environmental protection, safety and no side effects.
  • the invention adopts spray drying to quickly dry the raw material extract of the invention, and the obtained product has fine and uniform particle size, good fluidity, increased specific surface area, favorable dissolution in the body, and favorable absorption by the human body.
  • the raw material extract is spray-dried to obtain fine particulate matter. Since it contains a viscous substance such as sugar, protein or starch, the present invention adds a residue of the raw material to the residue of the raw material in the process of spray drying of the raw material extract, and the residue of the raw material.
  • the spray-dried raw material extract has strong binding force with the residue of the raw material, which can prevent the occurrence of sticking; at the same time, the raw material medicine is fully utilized, and no other external auxiliary materials are added. Conducive to the preservation of its natural biological activity.
  • the capsule prepared by the above preparation has safe and no side effects, does not destroy the biological active component of the original plant during the extraction process, and can fully prepare the biological active component into an extract having a large specific surface area, which is beneficial to dissolution in the body and absorption by the human body; It can avoid the sticking phenomenon caused by the production process.
  • the temperature of the vacuum cold soaking method is 10-40 ° C, and the pressure is -0.1 to -0.09 MPa;
  • the fluid extracted by the supercritical fluid is carbon dioxide, and the extraction temperature is 30-40 ° C;
  • the spray drying temperature is 40-80 ° C, the drying time is 3 ⁇ 60S; the raw material extract has a particle size of 10 ⁇ 60 ⁇ m;
  • the capsule shell is a natural plant capsule shell.
  • the preparation of the walnut extract capsules includes:
  • the supercritical fluid extraction fluid is carbon dioxide, the extraction temperature is 30-40 ° C;
  • step S3 taking the walnut residue obtained in step S2, pulverizing and grinding;
  • step S4 taking the walnut extract obtained in step S2, spray-drying, adding the crushed and ground walnut residue obtained in step S3 during the spray drying process to obtain a dried walnut extract;
  • the spray drying temperature is 40-80 °C, drying time is 3 ⁇ 60S;
  • the walnut extract has a particle size of 10 ⁇ 60 ⁇ m;
  • the preparation of the fragrant extract capsule comprises:
  • the supercritical fluid extraction fluid is carbon dioxide, the extraction temperature is 30 ⁇ 40 ° C;
  • step S3 taking the fragrant residue obtained in step S2, pulverizing and grinding;
  • step S4 taking the fragrant extract obtained in step S2, spray-drying, adding the pulverized fragrant residue obtained in step S3 during the spray drying to obtain a dried fragrant extract;
  • the temperature of the spray drying is 40 ⁇ 80°C, drying time is 3 ⁇ 60S;
  • the fragrant extract has a particle size of 10 ⁇ 60 ⁇ m;
  • the preparation of the capsule of the American ginseng extract includes:
  • step S3 taking the residue of American ginseng obtained in step S2, pulverizing and grinding;
  • step S4 taking the American ginseng extract obtained in step S2, spray-drying, and adding the pulverized and burned American ginseng residue obtained in step S3 to obtain a dried American ginseng extract;
  • the spray drying temperature is 40-80.
  • °C, drying time is 3 ⁇ 60S;
  • the ginseng extract has a particle size of 10 ⁇ 60 ⁇ m;
  • step S5 taking 1 to 5 parts of the American ginseng extract obtained in step S4, filling into a natural plant capsule shell to obtain an American ginseng extract capsule;
  • the preparation of the Dendrobium extract capsule comprises:
  • step S3 taking the residue of the sarcophagus obtained in step S2, pulverizing and grinding;
  • step S4 taking the extract of Dendrobium extract obtained in step S2, spray-drying, adding the crushed and ground Dendrobium residue obtained in step S3 during the spray drying process to obtain a dried Dendrobium extract; the temperature of the spray drying is 40-80 °C, drying time is 3 ⁇ 60S; the particle extract of the Dendrobium extract is 10 ⁇ 60 ⁇ m;
  • step S5 taking 1 to 10 parts of the extract of Dendrobium obtained in step S4, filling into the shell of the natural plant capsule to obtain the extract of Dendrobium extract;
  • the walnut is black walnut
  • the fragrant worm is the Asian Artemisia scoparia
  • the American ginseng is the semi-wild American ginseng
  • the sarcophagus is the wild sarcophagus.
  • the walnut extract capsule, the fragrant extract capsule, the American ginseng extract capsule and the sarcophagus extract capsule are separately prepared, on the one hand, the mutual interference of the four plants in the treatment process can be avoided, It is beneficial to ensure the quality of the product; on the other hand, it can be administered flexibly according to the needs during treatment, and is suitable for different types of patients.
  • Another object of the present invention is to provide a method for preparing a medicament for preventing or treating lupus erythematosus by using the preparation method for anti-lubricated erythematosus combined with immunization.
  • the combination of the anti-lubricated erythematosus combined with immunization of the invention has the advantages of synergistic effect on the one hand, can inhibit bacteria and bacteria, enhance the body immunity, and quickly and effectively treat the disease of lupus erythematosus; on the other hand, it can further enhance the body immunity and regulate The body environment and repair the original damaged immune function, restore the normal balance of the body, can prevent the recurrence of lupus erythematosus.
  • the organic active ingredient of the medicinal plant can be extracted to the greatest extent without using organic solvents and destructive conditions, and the side effects can be safe and no effective side effects.
  • the active ingredient is made into a particulate material having a large specific surface area, which is beneficial to human body absorption.
  • the residue of the raw material is added in the process of spray drying of the raw material extract, which is beneficial to the common operation of the anti-stick wall to solve the raw material extract in the production.
  • the sticking phenomenon that occurs during the process can also make full use of the drug substance and maintain its naturalness.
  • the anti-lubricated erythematosus combined immunization drug of the invention adopts a capsule form, and the administration is precise and convenient, and is favorable for preservation and good stability.
  • the walnut is selected from the group consisting of Black Walnut (also known as Juglans nigra L).
  • Fragrant wormwood is selected from the group consisting of woormwood (or Artemisia absinthium).
  • American ginseng is selected from semi-wild American ginseng.
  • Dendrobium is selected from wild sarcophagus.
  • the endothelium of the walnuts that have been screened and cleaned will have a clean walnut endothelium at 10 to 40 ° C and -0.1 to -0.09 MPa. It is softened by cold dip under reduced pressure. After the endothelium of the walnut is softened, the supercritical fluid extraction is carried out at 30-40 °C using a carbon dioxide fluid, and the walnut extract and the walnut residue are separated. The walnut residue is ground and ground and passed through a 200-300 mesh sieve. The walnut extract is sprayed onto the sieved walnut residue by spray drying at a spray drying temperature of 40 to 80 ° C and a drying time of 3 to 60 s to obtain a walnut extract having a particle size of 10 to 60 ⁇ m. One part of the walnut extract was taken and filled into the shell of the natural plant capsule according to a conventional capsule filling method to obtain a walnut extract capsule.
  • the whole grass of the scented sapphire which has been screened and cleaned, is softened and immersed in a clean eucalyptus whole grass at a temperature of 10 to 40 ° C and -0.1 to -0.09 MPa.
  • the supercritical fluid extraction was carried out at 30 to 40 ° C using a carbon dioxide fluid, and the fragrant extract and the fragrant worm residue were separated.
  • the fragrant slag residue is ground and pulverized and passed through a 200-300 mesh sieve.
  • the fragrant extract was sprayed onto the sieved fragrant sage residue by spray drying at a temperature of 40 to 80 ° C and a drying time of 3 to 60 s to obtain a fragrant extract having a particle size of 10 to 60 ⁇ m.
  • the fragrant extract is extracted into a natural plant capsule shell according to a conventional capsule filling method to obtain a fragrant extract capsule.
  • the ginseng of the American ginseng was cleaned and softened under reduced pressure at 10 to 40 ° C and -0.1 to -0.09 MPa. After the parametric softening of American ginseng, the supercritical fluid extraction was carried out at 30-40 °C using a carbon dioxide fluid, and the American ginseng extract and the American ginseng residue were separated. Take the American ginseng residue and grind it and pass it through a 200-300 mesh sieve. The American ginseng extract is spray-dried onto the sieved American ginseng residue by spray drying at a temperature of 40 to 80 ° C and a drying time of 3 to 60 ⁇ to obtain an American ginseng extract having a particle size of 10 to 60 ⁇ m. Take 1 to 5 parts of American ginseng extract and fill it into the natural plant capsule shell according to the conventional capsule filling method to obtain American ginseng extract capsule.
  • the sarcophagus of the cleaned sarcophagus was removed and softened under reduced pressure at 10 to 40 ° C and -0.1 to -0.09 MPa. After the stem of Dendrobium was softened, supercritical fluid extraction was carried out at 30-40 °C using a carbon dioxide fluid to separate the Dendrobium extract and Dendrobium residue.
  • the sarcophagus residue was ground and ground and passed through a 200-300 mesh sieve.
  • the Dendrobium extract is sprayed onto the sieved Dendrobium residue by spray drying at a temperature of 40 to 80 ° C for a drying time of 3 to 60 ⁇ to obtain a Dendrobium extract having a particle size of 10 to 60 ⁇ m. Take 1 to 10 parts of Dendrobium extract, and fill it into the shell of natural plant capsule according to the conventional capsule filling method to obtain the extract of Dendrobium extract.
  • the walnut extract capsule, the fragrant extract capsule, the American ginseng extract capsule, and the sarcophagus extract capsule are simultaneously administered in a certain ratio.
  • Test materials walnut extract, fragrant extract, American ginseng extract, and sarcophagus extract were all prepared in the same manner as in Example 1.
  • the extracts of walnut, fragrant extract, American ginseng extract and Dendrobium extract were respectively taken and washed in a water bath at 70 ° C for 1 hour, immersed for 12 hours, and concentrated for testing.
  • mice weighing 25-30 g were randomly divided into 5 groups according to their body weight. They were randomly divided into 5 groups according to their body weight, with 10 rats in each group, half male and half female.
  • the cyclophosphamide (cp) at a dose of 50 mg/kg ⁇ bw was used as a positive control, and the distilled water was used as a negative control.
  • the doses of the anti-lubricated erythematosus combined immunization drugs of the present invention were 2.00, 4.00, 6.00 g/kg ⁇ bw, respectively, with distilled water. Match to the desired concentration.
  • mice weighing 30-35 g were randomly divided into 5 groups.
  • the cyclophosphamide at a dose of 40 mg/kg ⁇ bw was used as a positive control, and the distilled water was used as a negative control.
  • the dose of the anti-lubricated lupus erythematosus combined with the immunization of the present invention was 2.00, 4.00, 6.00 g/kg ⁇ bw, and was adjusted to the desired concentration with distilled water. .
  • the rats were intragastrically administered once a day for 5 days. The animals were sacrificed 30 days after the last gavage. The epididymis tablets were taken and stained with eosin. Five animals were counted in each group.
  • the anti-lubricated erythematosus combined with immunization drug of the present invention did not significantly change the incidence of sperm abnormality in the mice, and there was no significant difference between the respective dose groups and the negative control group, while the cyclophosphamide positive control group had a difference. Extremely significant (P ⁇ 0.01). Therefore, the anti-lubricated erythematosus combined with immunization drug of the present invention affects sperm deformity in mice.
  • the anti-lubricated erythematosus combined immunization drug of the present invention prepared in the second to seventh embodiments was also subjected to a similar toxicological test, and the results were the same as those of the anti-lubricated erythematosus combined immunization drug of the present invention prepared in Example 1.
  • Test substance walnut extract capsule, fragrant extract capsule, American ginseng extract capsule, and sarcophagus extract capsule were all prepared by the first embodiment.
  • the recommended amount for oral administration in humans is 3.4 g/day, calculated as 60 kg body weight per person (one for each variety), equivalent to 0.057 g/kg ⁇ bw.
  • the equivalent dose of the mouse is equivalent to 10 times the recommended amount of the human body, that is, 0.57 styling product per day/kg ⁇ bw (medium dose).
  • One dose group was set up and down: 1.70 g of styling product/kg ⁇ bw (high dose) and 0.057 g of styling product/kg ⁇ bw (low dose). The samples were prepared in sterilized water and the control group was replaced with sterilized water.
  • DTH delayed type hypersensitivity test
  • mice were intraperitoneally injected with 2% (v/v, prepared with physiological saline) of SRBC (0.2 mL/mouse), and the thickness of the left and right ankles was measured 4 days after sensitization. The same site was measured three times and averaged. Then, 20% (v/v, prepared with physiological saline) of SRBC (20 ⁇ L/mouse) was injected subcutaneously at the measurement site, and the thickness of the left and right crotch portions was measured 24 hours after the injection, and the same portion was measured three times, and the average value was taken.
  • the degree of DTH is expressed by the difference in the thickness of the ankle before and after the injection (swelling degree of the foot).
  • mice were orally administered with different doses of the anti-lubricated erythematosus combined with the immunizing drug of the present invention for 30 days.
  • the swelling degree of the ankle was compared between the low, middle and high dose groups and the control group.
  • P ⁇ 0.01 the anti-lubricated erythematosus combined with immunization drug of the present invention can significantly enhance the delayed type hypersensitivity in mice.
  • mice were immunized by intraperitoneal injection of 0.2 mL of 2% (v/v, prepared with physiological saline) SRBC (0.2 mL/mouse). After 5 days, the mice were sacrificed, the spleens were taken, gently shredded, and a cell suspension was prepared using Hank's solution, washed, centrifuged twice, and the cells were suspended in 5 mL of Hank's solution. After heating and dissolving the surface medium (agarose), mix with an equal amount of double Hank's solution, dispense small tubes, 0.5 mL per tube, and add 50 ⁇ L of 10% (v/v, prepared with SA solution) to the tube.
  • SRBC physiological saline
  • mice were orally administered with different doses of the anti-lubricated erythematosus combined with the immunizing drug of the present invention for 30 days. After statistical analysis, there was no significant difference between the low, medium and high dose groups and the control group (P>0.05), that is, the anti-lub would be combined with the immunization drug of the present invention. There was no effect on the number of cells produced.
  • mice The tail vein of the mice was injected with 4 times diluted Indian ink in physiological saline, and 0.1 mL was injected per 10 g of body weight, and the ink was immediately counted immediately after the injection. Two or ten minutes after the ink was injected, 20 ⁇ L of blood was taken from the iliac venous plexus, and added to 2 mL of 0.1% Na 2 CO 3 solution, and shaken. The optical density value (OD) was measured at a wavelength of 600 nm with a 721 spectrophotometer using a 0.1% Na 2 CO 3 solution as a blank control. The mice were sacrificed, liver and spleen were taken, weighed (weight m, liver weight m 1 , spleen weight m 2 ), and the phagocytic index a was calculated as follows:
  • Phagocytic index Control 25 6.01 ⁇ 0.52 Low dose 15 6.33 ⁇ 0.40 Medium dose 14 5.89 ⁇ 0.48 High dose 14 5.77 ⁇ 0.43
  • mice were orally administered with different doses of the anti-lubricated erythematosus combined with the immunizing drug of the present invention for 30 days.
  • the swelling degree of the ankle was compared between the low, middle and high dose groups and the control group.
  • P>0.05 the anti-lubricated erythematosus combined with immunization drug of the present invention had no effect on the mononuclear-macrophage carbon clearance function of mice.
  • oral administration of different doses of the anti-lubricated erythematosus combined with the immunizing agent of the present invention for 30 days can significantly enhance the delayed type hypersensitivity of the mouse, and the number of antibody-producing cells in the mouse is small.
  • the mononuclear-macrophage carbon clearance function of the rat had no effect.
  • the anti-lubricated erythematosus combined with immunization drug of the present invention has an effect of enhancing cellular immune function.
  • the anti-lubricated erythematosus combined immunization drug of the present invention prepared in the second to seventh embodiments was also subjected to a similar immunomodulatory test, and the results were the same as those of the anti-lubricated erythematosus combined immunization drug of the present invention prepared in Example 1. .
  • the anti-lubricated erythematosus combined immunization drug of the present invention prepared in Example 1 was tested for coliform bacteria, total number of colonies, pathogenic bacteria, mold, and yeast.
  • a set of samples just produced and a set of samples placed at 25 ° C / 60% RH for 3 years were tested separately, and both sets of samples were samples of the same production batch.
  • the anti-lubris combined immunotherapy drug was placed for 3 years after production and at 25 ° C / 60% RH, and its hygiene test passed.
  • the anti-lubricated erythematosus combined immunization drug of the present invention prepared in Example 2 to Example 7 was also subjected to a similar hygienic test, and the results were the same as those of the anti-lubricated erythematosus combined immunization drug of the present invention prepared in Example 1.
  • Clinical trials were performed on patients with lupus erythematosus.
  • the criteria for inclusion were: pathological or cytological confirmation.
  • 4 or more of the following 11 criteria can be diagnosed as systemic lupus erythematosus: (1) butterfly erythema on the cheek; (2) discoid erythema; (3) sunlight; (4) oral ulcer; (5) non-erosive; (6) pleurisy, pericarditis; (7) proteinuria more than 0.5 grams per day; (8) abnormal nervous system: convulsions or mental illness; (9) hematological abnormalities, hemolytic, or less white blood cells At 4 ⁇ 109 / liter, or lymphocytes less than 1.5 ⁇ 109 / liter, or platelets less than 100 ⁇ 109 / liter; (10) immunological abnormalities, lupus cell positive, anti-double-stranded dna antibody positive or anti-sm antibody positive , or serum Test false positive; (11) immunofluorescence anti-nucle
  • a total of 200 patients with systemic lupus erythematosus were enrolled, including 30 males and 170 females, aged 20-60 years, with an average age of 30 years.
  • Therapeutic methods The whole group of patients were orally administered with the anti-lubricated lupus erythematosus combined with the immunization drug of the present invention every day, one for each capsule, one at a time each night, and six months for one course of treatment.

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Abstract

一种***和增强免疫力的药物，由以下原料制成（按重量份计）：胡桃萃取物（1），香艾萃取物（0.5-2），西洋参萃取物（1-5），石斛萃取物（1-10）。所述的药物是通过超临界流体萃取和喷雾干燥等方法制备的。所述药物能增强免疫力，预防或***。

Description

一种抗红斑狼疮联合免疫用药物及其制备方法、用途 技术领域
本发明属于药物领域，更具体地说，它涉及一种抗红斑狼疮联合免疫用药物及其制备方法、用途。
背景技术
红斑狼疮(Lupus Erythematosus，简称LE)，为自身免疫性***病。红斑狼疮分为盘状红斑狼疮(DLE)、亚急性皮肤型红斑狼疮(SCLE)、***性红斑狼疮(SLE)、深在性红斑狼疮(LEP)、新生儿红斑狼疮(NLE)、药物性红斑狼疮(DIL)等亚型。目前对红斑狼疮的病因尚未完全了解，但医学界普遍认为红斑狼疮的病因可能与遗传、性激素、环境等多种因素作用造成机体免疫功能紊乱有关。现有技术中，***主要有西医西药疗法、中医疗法。西医西药***有一定的效果，特别是疾病的爆发期或急性活动期，应用大剂量的激素及环磷酰胺等药物对迅速控制病情有积极作用，但长期大量应用这些药物，会产生副作用，对机体造成损害，甚至危及生命，同时易反复发作。采用中医***，治疗周期较长，皮肤、内脏等多种组织和器官受损较广，影响患者治愈的信心，导致其治疗效果一般。
宋晓萍等在皮肤红斑狼疮的中西医治疗进展与思考(《海南医学》2010年第21卷第21期)中，提出了皮肤红斑狼疮的中西医治疗法。但西药和中药的配伍性还待研究，且中、西医结合疗法，主要是通过加强患者的免疫力，结合西药来达到治疗的目的，这种方法只能加强患者在治疗期间的免疫力，而无法修复其治疗前被破坏的免疫***，无法保证机体内环境的免疫力平衡性，这将导致疾病容易复发；两种治疗之间易互相干扰，容易发生副反应。
如何快速、有效、安全的***，并防止其反复发作，已成为行业内重点研究的方向之一。
发明内容
针对现有技术存在的不足，本发明的目的在于提供一种能抑菌杀菌，增强机体免疫力，快速有效的***这种疾病；能进一步增强机体免疫力，调节机体内环境，恢复机体的正常平衡，能防止红斑狼疮的复发的抗红斑狼疮联合免疫用药物。
为实现上述目的，本发明提供了如下技术方案：
一种抗红斑狼疮联合免疫用药物，所述抗红斑狼疮联合免疫用药物由胡桃萃取物、香艾萃 取物、西洋参萃取物、石斛萃取物为原料；所述胡桃萃取物、香艾萃取物、西洋参萃取物和石斛萃取物的重量配比为1∶(0.5～2)∶(1～5)∶(1～10)。
作为优选，所述胡桃为黑胡桃，香艾为中亚苦蒿，西洋参为半野生西洋参，石斛为野生石斛。
胡桃，为胡桃科植物胡桃Julans regia L.。本发明的胡桃萃取物是用北美洲黑胡桃(Black Walnut，Juglans nigra L)的内皮萃取制成，其含有胡桃醌、黄酮类物质、鞣质、多糖、没食子酸，挥发油、植物粘胶、白蛋白、矿物质、纤维素、亚油酸、甘油酯、亚麻酸、油酸甘油酯、核黄素、胡萝卜素、水杨梅丁素、木麻黄素、刺玫果素，研究发现其活性成分的比例不同均能达到本发明的目的，且通过本发明的制备方法制备的胡桃萃取物的效果最佳。
香艾，又名艾叶、艾蒿、蕲艾或家艾，为菊科植物艾Artemisia argyi Levl.et Vant.的干燥叶。其辛、苦，温；有小毒，能散寒止痛，温经止血。本发明的香艾萃取物优选用北美洲中亚苦蒿(woormwood，或Artemisia absinthium)的全草萃取制成，其主要成分为挥发油，油中有萜品烯醇-4(Terpinenol-4)、β-石竹烯(β-Caryophyllene)、蒿醇(Artemisia alcohol)、芳樟醇(Linalool)、樟脑(Camphorae)、龙脑(冰片，Borneol)、桉油素(Cineol，Eucalyptol)及反式-葛缕醇、α-萜品烯醇等，又含多糖类物质、鞣酸、侧柏酮、水芹烯、β-谷甾醇、5，7-二羟基-6，3′4′-三甲基黄酮，研究发现其活性成分的比例不同均能达到本发明的目的，且通过本发明的制备方法制备的香艾萃取物的效果最佳。
西洋参，又名花旗参、洋参、美国参，其为五加科人参属西洋参Panax quinquefolium Linn的根，其为中国东北盛产的人参Panx ginseng同科同属。其性寒，有养阴补气、降压和解热作用。半野生西洋参为西洋参的美国花旗参的品种之一，其主要产地为威斯康辛州、纽约州、肯塔基州等25个州，其为将西洋参的种子放在类似野生的环境培育得到。本发明的西洋参萃取物是用半野生西洋参的参体萃取制成，其主要成分为人参皂苷、挥发油、油脂中脂肪酸、有机酸、氨基酸、微量元素、碳水化合物、果胶、甾醇，人参皂苷主要有人参皂苷Ro、Rb1、Rb2、Rb3、Rc、Rd、Re、Rf、Rg1、Rg2、Rg3、Rh1、RAO、西洋参皂苷R1、绞股蓝皂苷XI、七叶胆苷XVII、假人参皂苷主要为F11、人参皂F2、F3等，挥发油主要有β-金合欢烯、倍半萜类化合物、没药烯、α-姜黄烯等，研究发现其活性成分的比例不同均能达到本发明的目的，且通过本发明的制备方法制备的西洋参萃取物的效果最佳。
石斛，别名为吊兰、杜兰，其为兰科植物金钗石斛Dendrobium nobile Lindl.、铁皮石斛Dendrobium candidum Wall.ex Lindl.或马鞭石斛Dendrobium fimbriatum Hook.Var. oculatum Hook及其近似种的茎。其甘，微寒，益胃生津，滋阴清热。本发明的石斛萃取物是用野生石斛的茎萃取制成，其含多种生物碱、多糖、氨基酸、粘液质、淀粉、菲类及联苄类等，生物碱包括石斛碱(Dendrobine)、石斛副碱(Dendrine)、石斛胺碱(Dendramine)、石斛星碱(Dendroxine)、6-羟基石斛星碱(6-Hydroxy-dendroxine)等，研究发现其活性成分的比例不同均能达到本发明的目的，且通过本发明的制备方法制备的石斛萃取物的效果最佳。
研究发现，本发明的抗红斑狼疮联合免疫用药物，其胡桃萃取物、香艾萃取物、西洋参萃取物和石斛萃取物的重量配比为1∶(0.5～2)∶(1～5)∶(1～10)，其组合增效：
(1)对红斑狼疮患者起到缓解和改善症状的左右，如缓解疼痛、祛除瘙痒、增加食欲、镇痛补血、生血、保肝、保持血糖稳定；
(2)能抑菌杀菌、抗病毒、排毒、抗炎，具有较强的抗氧化性和羟自由基清除能力，增强红斑狼疮患者对抗其他新疾病的能力；
(3)其他组分结合其富含的多糖使其具有增强机体免疫力的作用，对机体体液免疫功能、细胞免疫功能的促进和诱生多种细胞因子，能快速有效的***；
(4)促进血液活力、调节中枢神经、保护心血管***，增强机体排毒功能，同时能增强机体免疫力和精力，提供机体营养所需，增强细胞免疫活性，调节机体内环境并修复原被破坏的免疫机能，恢复正常的生理平衡，防止红斑狼疮的复发。
研究发现，优选所述胡桃萃取物、香艾萃取物、西洋参萃取物和石斛萃取物的重量配比为1∶1∶5∶10。
作为优选，所述药物的制剂为胶囊剂。
以胶囊剂给药，有利于保证本发明的抗红斑狼疮联合免疫用药物能准确、方便给药，有利于控制。
本发明的目的还在于提供一种所述的抗红斑狼疮联合免疫用药物的制备方法，所述方法包括胡桃萃取物胶囊的制备、香艾萃取物胶囊的制备、西洋参萃取物胶囊的制备、石斛萃取物胶囊的制备；所述胡桃萃取物胶囊、香艾萃取物胶囊的、西洋参萃取物胶囊、石斛萃取物胶囊的制备均包括：
S1，取原料进行筛选和清洗，将干净的原料通过减压冷浸法软化；
S2，取经S1步软化的原料，超临界流体萃取，分离得到萃取液与残渣；
S3，取S2步得到的残渣，将其粉碎研磨；
S4，取S2步得到的萃取液，将其喷雾干燥，喷雾干燥的过程中加入S3步得到的粉碎研磨后的残渣，得到干燥的原料萃取物；
S5，取配方量的S4步得到的原料萃取物，填充至胶囊壳内，得到胶囊。
本发明采用减压冷浸软化法，借负压的作用将水迅速吸入，加速药材的软化，确保在常温下用水软化药材，缩短浸润时间，减少有效成分的流失、破坏和药材的霉变。
本发明采用超临界流体萃取技术萃取药用植物，其超临界流体萃取的流体为二氧化碳，其萃取温度为30～40℃，能避免挥发油等成分在萃取过程中的挥发，能最大程度的保持药用植物的生物活性成分；同时因为过程中不适用有机溶剂，无溶剂残留，保证了原料的纯天然活性，环保，安全无副作用。
本发明采用喷雾干燥将本发明的原料萃取液快速干燥，得到的产品粒度细小、均匀，流动性好，增加了其比表面积，有利于其在机体内的溶出，有利于人体吸收。原料萃取液经喷雾干燥后得到细小的颗粒物质，由于其含有糖类、蛋白质、淀粉等黏性较大的物质，本发明在原料萃取液喷雾干燥的过程中加入其原料的残渣，原料的残渣能吸附经喷雾干燥的原料萃取液，且经喷雾干燥的原料萃取液与原料的残渣的结合力较强，能防止其出现粘壁的情况；同时充分利用原料药，未添加其他外加辅料，有利于保存其纯天然生物活性。
通过上述制备得到的胶囊，安全无副作用，提取过程无破坏原植物生物活性成分，还能充分将其生物活性成分制成比表面积大的萃取物，有利于体内溶出和人体吸收；同时使用该方法，能避免生产过程中产生的粘壁现象等。
研究发现，优选S1中，所述减压冷浸法的温度为10～40℃，压强为-0.1～-0.09MPa；
S2中，所述超临界流体萃取的流体为二氧化碳，萃取温度为30～40℃；
S4中，所述喷雾干燥的温度为40～80℃，干燥时间为3～60S；所述原料萃取物的颗粒度为10～60μm；
S5中，所述胶囊壳为天然植物胶囊壳。
本发明的目的还在于提供一种所述的抗红斑狼疮联合免疫用药物的制备方法，所述方法包括胡桃萃取物胶囊的制备、香艾萃取物胶囊的制备、西洋参萃取物胶囊和石斛萃取物胶囊的制备；
所述胡桃萃取物胶囊的制备包括：
S1，取胡桃筛选、清洗，将干净的胡桃的内皮通过减压冷浸法软化；所述减压冷浸法的温度为10～40℃，压强为-0.1～-0.09MPa；
S2，取经S1步软化的胡桃，超临界流体萃取，分离得到胡桃萃取液与胡桃残渣；所述超临界流体萃取的流体为二氧化碳，萃取温度为30～40℃；
S3，取S2步得到的胡桃残渣，将其粉碎研磨；
S4，取S2步得到的胡桃萃取液，将其喷雾干燥，喷雾干燥的过程中加入S3步得到的粉碎研磨后的胡桃残渣，得到干燥的胡桃萃取物；所述喷雾干燥的温度为40～80℃，干燥时间为3～60S；所述胡桃萃取物的颗粒度为10～60μm；
S5，取1份S4步得到的胡桃萃取物，填充至天然植物胶囊壳内，得到胡桃萃取物胶囊；所述香艾萃取物胶囊的制备包括：
S1，取香艾筛选、清洗，将干净的香艾的全草通过减压冷浸法软化；所述减压冷浸法的温度为10～40℃，压强为-0.1～-0.09MPa；
S2，取经S1步软化的香艾，超临界流体萃取，分离得到香艾萃取液与香艾残渣；所述超临界流体萃取的流体为二氧化碳，萃取温度为30～40℃；
S3，取S2步得到的香艾残渣，将其粉碎研磨；
S4，取S2步得到的香艾萃取液，将其喷雾干燥，喷雾干燥的过程中加入S3步得到的粉碎研磨后的香艾残渣，得到干燥的香艾萃取物；所述喷雾干燥的温度为40～80℃，干燥时间为3～60S；所述香艾萃取物的颗粒度为10～60μm；
S5，取54份S4步得到的香艾萃取物，填充至天然植物胶囊壳内，得到香艾萃取物胶囊；所述西洋参萃取物胶囊的制备包括：
S1，取西洋参筛选、清洗，将干净的西洋参的参体通过减压冷浸法软化；所述减压冷浸法的温度为10～40℃，压强为-0.1～-0.09MPa；
S2，取经S1步软化的西洋参，超临界流体萃取，分离得到西洋参萃取液与西洋参残渣；所述超临界流体萃取的流体为二氧化碳，萃取温度为30～40℃；
S3，取S2步得到的西洋参残渣，将其粉碎研磨；
S4，取S2步得到的西洋参萃取液，将其喷雾干燥，喷雾干燥的过程中加入S3步得到的粉碎研磨后的西洋参残渣，得到干燥的西洋参萃取物；所述喷雾干燥的温度为40～80℃，干燥时间为3～60S；所述西洋参萃取物的颗粒度为10～60μm；
S5，取1～5份S4步得到的西洋参萃取物，填充至天然植物胶囊壳内，得到西洋参萃取物胶囊；
所述石斛萃取物胶囊的制备包括：
S1，取石斛筛选、清洗，将干净的石斛茎叶通过减压冷浸法软化；所述减压冷浸法的温度为10～40℃，压强为-0.1～-0.09MPa；
S2，取经S1步软化的石斛，超临界流体萃取，分离得到石斛萃取液与石斛残渣；所述超临界流体萃取的流体为二氧化碳，萃取温度为30～40℃；
S3，取S2步得到的石斛残渣，将其粉碎研磨；
S4，取S2步得到的石斛萃取液，将其喷雾干燥，喷雾干燥的过程中加入S3步得到的粉碎研磨后的石斛残渣，得到干燥的石斛萃取物；所述喷雾干燥的温度为40～80℃，干燥时间为3～60S；所述石斛萃取物的颗粒度为10～60μm；
S5，取1～10份S4步得到的石斛萃取物，填充至天然植物胶囊壳内，得到石斛萃取物胶囊；
所述胡桃为黑胡桃，香艾为中亚苦蒿，西洋参为半野生西洋参，石斛为野生石斛。
本发明的抗红斑狼疮联合免疫用药物中，胡桃萃取物胶囊、香艾萃取物胶囊、西洋参萃取物胶囊和石斛萃取物胶囊分开制备，一方面可以避免四种植物在处理过程中的相互干扰，有利于保证产品质量；另一方面在治疗时，可以根据需求而灵活给药，适于不同类型的病人。
本发明的目的还在于提供一种所述的抗红斑狼疮联合免疫用药物用以制备预防或***的药物的用途。
本发明的目的还在于提供一种所述的制备方法制备得到的抗红斑狼疮联合免疫用药物用以制备预防或***的药物的用途。
通过采用上述技术方案，具有以下有益效果：
1.本发明的抗红斑狼疮联合免疫用药物组合增效，一方面能抑菌杀菌，增强机体免疫力，快速有效的***这种疾病；另一方面其能进一步增强机体免疫力，调节机体内环境并修复原被破坏的免疫机能，恢复机体的正常平衡，能防止红斑狼疮的复发。
2.本发明的抗红斑狼疮联合免疫用药物的制备过程中，未使用有机溶剂和破坏性条件，能最大程度的提取药用植物的生物活性成分，安全无副作用，同时还能将其有效生物活性成分制成比表面积大的颗粒物质，有利于人体吸收。
3.本发明的抗红斑狼疮联合免疫用药物的制备过程中，在原料萃取液喷雾干燥的过程中加入其原料的残渣，有利于和常规防止粘壁的操作共同作用来解决原料萃取物在生产的过程中出现的粘壁现象，还能充分利用原料药，保持纯天然性。
4.本发明的抗红斑狼疮联合免疫用药物采用胶囊形式，给药精确且方便，有利于保存，稳定性好。
具体实施方式
实施例中，胡桃选自北美洲黑胡桃(Black Walnut，又名Juglans nigra L)。香艾选自中亚苦蒿(woormwood，或Artemisia absinthium)。西洋参选自半野生西洋参。石斛选自野生石斛。
取经筛选、清洗的胡桃的内皮，将干净的胡桃的内皮在10～40℃和-0.1～-0.09MPa 下减压冷浸软化。胡桃的内皮软化后，在30～40℃下利用二氧化碳流体进行超临界流体萃取，分离得到胡桃萃取液与胡桃残渣。取胡桃残渣粉碎研磨，过200-300目筛。利用喷雾干燥将胡桃萃取液喷涂到经过筛的胡桃残渣上，喷雾干燥的温度为40～80℃，干燥时间为3～60S，得到颗粒度为10～60μm的胡桃萃取物。取1份胡桃萃取物，按常规的胶囊填充方法填充至天然植物胶囊壳内，得到胡桃萃取物胶囊。
取经筛选、清洗的香艾的全草，将干净的香艾的全草在10～40℃和-0.1～-0.09MPa下减压冷浸软化。香艾软化后，在30～40℃下利用二氧化碳流体进行超临界流体萃取，分离得到香艾萃取液与香艾残渣。取香艾残渣粉碎研磨，过200-300目筛。利用喷雾干燥将香艾萃取液喷涂到经过筛的香艾残渣上，喷雾干燥的温度为40～80℃，干燥时间为3～60S，得到颗粒度为10～60μm的香艾萃取物。取香艾萃取物，按常规的胶囊填充方法填充至天然植物胶囊壳内，得到香艾萃取物胶囊。
取经筛选、清洗的西洋参，将干净的西洋参的参体在10～40℃和-0.1～-0.09MPa下减压冷浸软化。西洋参的参体软化后，在30～40℃下利用二氧化碳流体进行超临界流体萃取，分离得到西洋参萃取液与西洋参残渣。取西洋参残渣粉碎研磨，过200-300目筛。利用喷雾干燥将西洋参萃取液喷涂到经过筛的西洋参残渣上，喷雾干燥的温度为40～80℃，干燥时间为3～60S，得到颗粒度为10～60μm的西洋参萃取物。取1～5份西洋参萃取物，按常规的胶囊填充方法填充至天然植物胶囊壳内，得到西洋参萃取物胶囊。
取经筛选、清洗的石斛，将干净的石斛的茎在10～40℃和-0.1～-0.09MPa下减压冷浸软化。石斛的茎软化后，在30～40℃下利用二氧化碳流体进行超临界流体萃取，分离得到石斛萃取液与石斛残渣。取石斛残渣粉碎研磨，过200-300目筛。利用喷雾干燥将石斛萃取液喷涂到经过筛的石斛残渣上，喷雾干燥的温度为40～80℃，干燥时间为3～60S，得到颗粒度为10～60μm的石斛萃取物。取1～10份石斛萃取物，按常规的胶囊填充方法填充至天然植物胶囊壳内，得到石斛萃取物胶囊。
使用本发明的抗红斑狼疮联合免疫用药物时，将胡桃萃取物胶囊、香艾萃取物胶囊、西洋参萃取物胶囊和石斛萃取物胶囊按一定比例同时给药。
表1本发明的抗红斑狼疮联合免疫用药物
Figure PCTCN2016078649-appb-000001
对比例一毒理学试验
受试物：胡桃萃取物、香艾萃取物、西洋参萃取物、石斛萃取物均由实施例一制备。分别取胡桃萃取物、香艾萃取物、西洋参萃取物、石斛萃取物，分别经70℃水浴1小时、浸泡12小时、浓缩后，供试验用。
2、试验动物：
选用中国药品生物制品检定所实验动物中心提供的昆明种健康小白鼠。
3、小鼠骨髓嗜多染红细胞微核试验
采用间隔24hr两次经口灌胃法进行试验。用体重25-30克小白鼠50只，按体重随机分为5组，按体重随机分为5组，每组10只，雌雄各半。以50mg/kg·bw剂量的环磷酰胺(cp)为阳性对照，蒸馏水为阴性对照，本发明的抗红斑狼疮联合免疫用药物的剂量分别为2.00、4.00、6.00g/kg·bw，用蒸馏水配至所需浓度。末次给本发明的抗红斑狼疮联合免疫用药物后6hr，颈椎脱臼处死动物，取胸骨骨髓用小牛血清稀释涂片，甲醇固定，Giemsa染色。 在光学显微镜下，每只动物计数1000个嗜多染红细胞(PRC)，微核的发生率以含微核的PRC千分率计，并进行统计处理。本发明的抗红斑狼疮联合免疫用药物对小鼠骨髓微核发生率的影响的结果统计见表2。
表2小鼠骨髓嗜多染红细胞微核试验的结果统计
Figure PCTCN2016078649-appb-000002
*与阴性对照组比较差异有极显著性(P＜0.01)。
由表2可见，小本发明的抗红斑狼疮联合免疫用药物各剂量组微核率，各剂量组与阴性对照组比较差异无显著性，而环磷酰胺组与阴性对照组比较差异有极显著性(P＜0.01)。未见本发明的抗红斑狼疮联合免疫用药物对小鼠骨髓嗜多染红细胞微核率产生影响。
4、小鼠***畸形试验
用体重30-35克的性成熟雄性小鼠50只，随机分为5组。以40mg/kg·bw剂量的环磷酰胺为阳性对照，蒸馏水为阴性对照，本发明的抗红斑狼疮联合免疫用药物剂量为2.00、4.00、6.00g/kg·bw，用蒸馏水配至所需浓度。每日灌胃一次，连续5天，末次灌胃后30天处死动物，取附睾制片，伊红染色，每组计数5只动物，每只动物计数1000个结构完整的***，计算畸变***发生率(以百分率计)，并进行统计处理。本发明的抗红斑狼疮联合免疫用药物对小鼠***畸形发生率的影响的结果统计见表3。
表3小鼠***畸形试验的结果统计
剂量(g/kg.bw) 动物数(只) 受检***数(个) ***畸形数(个) ***畸形率(％)
0.00 5 5000 81 1.62
2.00 5 5000 94 1.88
4.00 5 5000 76 1.52
6.00 5 5000 87 1.74
40mg/kg.bw(cp) 5 5000 480 9.6*
*与阴性对照组比较差异有极显著性(P＜0.01)
由表3可见，本发明的抗红斑狼疮联合免疫用药物对小鼠***畸形发生率未产生明显改变，各剂量组与阴性对照组比较差异无显著性，而环磷酰胺阳性对照组比较差异有极显著性(P＜0.01)。因此，本发明的抗红斑狼疮联合免疫用药物对小鼠***畸形产生影响。
对实施例二至实施例七制备的本发明的抗红斑狼疮联合免疫用药物也进行相似的毒理学试验，其结果和实施例一制备的本发明的抗红斑狼疮联合免疫用药物的结果相同。
对比例二免疫调节作用试验
1、受试物：胡桃萃取物胶囊、香艾萃取物胶囊、西洋参萃取物胶囊、石斛萃取物胶囊均由实施例一制备。人体口服推荐量为3.4g/天，以每人60kg体重计算(每个品种各一粒)，相当于0.057g/kg·bw。小鼠的等效剂量相当于人体推荐量的10倍，即每日0.57定型产品/kg·bw(中剂量)。上、下各设一个剂量组：1.70g定型产品/kg·bw(高剂量)和0.057g定型产品/kg·bw(低剂量)。样品以已消毒的水配制，对照组以已消毒的水代替。连续灌胃30天后测各项免疫指标。因为胡桃萃取物胶囊、香艾萃取物胶囊、西洋参萃取物胶囊、石斛萃取物胶囊均不溶于水，用10倍于推荐量的热水(70-80℃)分3次，每次1小时浸泡，然后在旋转蒸发仪(50-60℃)蒸发浓缩成30倍推荐剂量浓度，再按比例稀释进行灌胃。
2、试验动物：
选用中国药品生物制品检定所实验动物中心提供的昆明种健康雄性小鼠75只，体重18-22g，分为4组，剂量组每组15只，对照组30只，进行迟发型***反应和抗体生成细胞检测。
选用中国药品生物制品检定所实验动物中心提供的昆明种健康雌性小鼠75只，体重18-22g，分为4组，剂量组每组15只，对照组30只，进行碳粒廓清试验。
3、迟发型***反应检测(DTH)(足跖增厚法)
小鼠经腹腔注射2％(v/v，用生理盐水配制)的SRBC(0.2mL/每鼠)，致敏后4天，测量左右足跖部厚度，同一部位测量三次，取平均值。然后在测量部位皮下注射20％(v/v，用生理盐水配制)的SRBC(20μL/每鼠)，于注射后24小时测量左右跖部厚度，同一部位测量三次，取平均值。以注射前后足跖厚度的差值(足跖肿胀度)来表示DTH的程度。本发明的抗红斑狼疮联合免疫用药物对小鼠迟发型***反应(DTH)的影响的结果统计见表4。
表4迟发型***反应检测(DTH)的结果统计
组别 动物数(只) 足跖肿胀度(mm)
对照 30 0.573±0.109
低剂量 15 0.663±0.106
中剂量 15 0.712±0.105
高剂量 15 0.700±0.102
由表4可见，经口给予小鼠不同剂量的本发明的抗红斑狼疮联合免疫用药物30天，经统计学处理，其足跖肿胀度在低、中、高剂量组与对照组间比较均有极显著性差异(P＜0.01)，即本发明的抗红斑狼疮联合免疫用药物能明显增强小鼠的迟发型***反应。
4、抗体生成细胞检测(Jerne改良玻片法)
小鼠经腹腔注射0.2mL 2％(v/v，用生理盐水配制)的SRBC(0.2mL/每鼠)进行免疫。5天后，将小鼠处死，取脾，轻轻撕碎，用Hank′s液制成细胞悬液，洗涤、离心2次，将细胞悬浮在5mL Hank′s液上。将表层培养基(琼脂糖)加热溶解后，与等量双倍Hank′s液混合，分装小试管、每管0.5mL，再向管内加50μL 10％(v/v，用SA液配制)的SRBC、脾细胞悬液20μL，迅速混匀后，倾倒于己刷琼脂糖薄层的玻片上，待琼脂凝固后，将玻片水平扣放在片架上，放二氧化碳培养箱中温育1.5hr，然后用SA缓冲液稀释的补体(1∶10)加到玻片架凹槽内，继续温育1.5hr后，计数溶血空斑数。本发明的抗红斑狼疮联合免疫用药物对抗体生成细胞数的影响的结果统计见表5。
表5抗体生成细胞检测的结果统计
组别 动物数(只) 溶血空斑数(×103/全脾)
对照 22 115.9±50.5
低剂量 11 108.2±43.6
中剂量 13 141.6±51.3
高剂量 12 129.6±101.3
由表5可见，经口给予小鼠不同剂量的本发明的抗红斑狼疮联合免疫用药物30天， 经统计学处理，其足跖肿胀度在低、中、高剂量组与对照组间比较均无极显著性差异(P＞0.05)，即本发明的抗红斑狼疮联合免疫用药物对小鼠的抗体生成细胞数无影响。
5、小鼠碳廓清实验测试
小鼠尾静脉注射以生理盐水稀释4倍的印度墨汁，每10g体重注射0.1mL，墨汁注入后立即计时。注入墨汁后2、10min，分别从内眦静脉丛取血20μL，加到2mL 0.1％Na2CO3溶液中，摇匀。以0.1％Na2CO3溶液作空白对照，用721分光光度计在600nm波长处测光密度值(OD)。将小鼠处死，取肝、脾，称重(体重为m，肝重为m1，脾重为m2)，按下式计算吞噬指数a：
Figure PCTCN2016078649-appb-000003
Figure PCTCN2016078649-appb-000004
本发明的抗红斑狼疮联合免疫用药物对对小鼠单核-巨噬细胞吞噬功能的影响的结果统计见表6。
表6小鼠碳廓清实验测试的结果统计
组别 动物数(只) 吞噬指数
对照 25 6.01±0.52
低剂量 15 6.33±0.40
中剂量 14 5.89±0.48
高剂量 14 5.77±0.43
由表6可见，经口给予小鼠不同剂量的本发明的抗红斑狼疮联合免疫用药物30天，经统计学处理，其足跖肿胀度在低、中、高剂量组与对照组间比较均无极显著性差异(P＞0.05)，即本发明的抗红斑狼疮联合免疫用药物对小鼠单核-巨噬细胞碳廓清功能无影响。
由表4至表6可见，经口给予小鼠不同剂量的本发明的抗红斑狼疮联合免疫用药物30天，能明显增强小鼠的迟发型***反应，对小鼠的抗体生成细胞数和小鼠单核-巨噬细胞碳廓清功能无影响。由此可见，本发明的抗红斑狼疮联合免疫用药物具有增强细胞免疫功能的作用。
对实施例二至实施例七制备的本发明的抗红斑狼疮联合免疫用药物也进行相似的免疫调节作用试验，其结果和实施例一制备的本发明的抗红斑狼疮联合免疫用药物的结果相同。
对比例三、制剂的卫生学检验
参考GB4789-94和GB4789.15-94，对实施例一制备的本发明的抗红斑狼疮联合免疫用药物进行大肠菌群、菌落总数、致病菌、霉菌、酵母计数的检测。分别对刚生产的一组样品和在25℃/60％RH下放置3年后的一组样品进行检测，且两组样品均为同一个生产批次的样品。每组样品均平行取10个样品。其卫生学检验结果详见表7。
表7卫生学检验结果统计
Figure PCTCN2016078649-appb-000005
由表7可见，抗红斑狼疮联合免疫用药物在生产后及在25℃/60％RH下放置3年，其卫生学检验合格。
对实施例二至实施例七制备的本发明的抗红斑狼疮联合免疫用药物也进行相似的卫生学检验，其结果和实施例一制备的本发明的抗红斑狼疮联合免疫用药物的结果相同。
对比例四、临床试验
取红斑狼疮患者病例进行临床试验，病例入选标准为：均经病理学或细胞学证实诊断。其中，以下11项标准中符合4项及以上者，即可诊断为***性红斑狼疮：(1)面颊部蝶形红斑；(2)盘状红斑；(3)日光；(4)口腔溃疡；(5)非侵蚀性；(6)胸膜炎、心包炎；(7)蛋白尿每日0.5克以上；(8)神经***异常：抽搐或精神病；(9)血液学异常，溶血性，或白细胞少于4×109/升，或淋巴细胞少于1.5×109/升，或血小板少于100×109/升；(10)免疫学异常，狼疮细胞阳性，抗双链dna抗体阳性或抗sm抗体阳性，或血清 试验假阳性；(11)免疫荧光抗核抗体阳性。其检查手段包括血常规检查(包括血色素、红细胞、白细胞、血小板)、尿常规检查和血液生化检查。
共取***性红斑狼疮患者200例，其中男30例，女170例，年龄20-60岁，平均年龄30岁。
治疗方法：全组病例每天均口服实施例一制备的本发明的抗红斑狼疮联合免疫用药物，每次每种胶囊各一个，每日晚各一次，6个月为一个疗程。
治疗后，在2个月内完全治愈的有25例，在2-4个月内完全治愈的有44例，在4-6个月内完全治愈的有61例。服完一个疗程后，已治愈的130例中，有106例在2年内均未发生复发的情况，而其余24例复发时症状较之前较轻。在这24例复发时，继续服用本发明的抗红斑狼疮联合免疫用药物一个疗程，其均在2个月内完全治愈。
毒副反应观察：全组病例在治疗期间均为观察到心、肝、肾功能等的不良反应，外周血象未见异常。未观察到皮疹等过敏现象发生。
以上所述仅是本发明的优选实施方式，本发明的保护范围并不仅局限于上述实施例，凡属于本发明思路下的技术方案均属于本发明的保护范围。应当指出，对于本技术领域的普通技术人员来说，在不脱离本发明原理前提下的若干改进和润饰，这些改进和润饰也应视为本发明的保护范围。

Claims (9)

  1. 一种抗红斑狼疮联合免疫用药物，其特征在于，所述抗红斑狼疮联合免疫用药物由胡桃萃取物、香艾萃取物、西洋参萃取物、石斛萃取物为原料；所述胡桃萃取物、香艾萃取物、西洋参萃取物和石斛萃取物的重量配比为1∶(0.5～2)∶(1～5)∶(1～10)。
  2. 根据权利要求1所述的抗红斑狼疮联合免疫用药物，其特征在于，所述胡桃为黑胡桃，香艾为中亚苦蒿，西洋参为半野生西洋参，石斛为野生石斛。
  3. 根据权利要求1所述的抗红斑狼疮联合免疫用药物，其特征在于，所述胡桃萃取物、香艾萃取物、西洋参萃取物和石斛萃取物的重量配比为1∶1∶5∶10。
  4. 根据权利要求1或2或3所述的抗红斑狼疮联合免疫用药物，其特征在于，所述药物的制剂为胶囊剂。
  5. 一种如权利要求4所述的抗红斑狼疮联合免疫用药物的制备方法，其特征在于，所述方法包括胡桃萃取物胶囊的制备、香艾萃取物胶囊的制备、西洋参萃取物胶囊的制备、石斛萃取物胶囊的制备；所述胡桃萃取物胶囊、香艾萃取物胶囊、西洋参萃取物胶囊、石斛萃取物胶囊的制备均包括：
    S1，取原料进行筛选和清洗，将干净的原料通过减压冷浸法软化；
    S2，取经S1步软化的原料，超临界流体萃取，分离得到萃取液与残渣；
    S3，取S2步得到的残渣，将其粉碎研磨；
    S4，取S2步得到的萃取液，将其喷雾干燥，喷雾干燥的过程中加入S3步得到的粉碎研磨后的残渣，得到干燥的原料萃取物；
    S5，取配方量的S4步得到的原料萃取物，填充至胶囊壳内，得到胶囊。
  6. 根据权利要求5所述的抗红斑狼疮联合免疫用药物的制备方法，其特征在于，
    S1中，所述减压冷浸法的温度为10～40℃，压强为-0.1～-0.09MPa；
    S2中，所述超临界流体萃取的流体为二氧化碳，萃取温度为30～40℃；
    S4中，所述喷雾干燥的温度为40～80℃，干燥时间为3～60S；所述原料萃取物的颗粒度为10～60μm；
    S5中，所述胶囊壳为天然植物胶囊壳。
  7. 一种如权利要求2所述的抗红斑狼疮联合免疫用药物的制备方法，其特征在于，所述方法包括胡桃萃取物胶囊的制备、香艾萃取物胶囊的制备、西洋参萃取物胶囊和石斛萃取物胶囊的制备；
    所述胡桃萃取物胶囊的制备包括：
    S1，取胡桃筛选、清洗，将干净的胡桃的内皮通过减压冷浸法软化；所述减压冷浸法的温度为10～40℃，压强为-0.1～-0.09MPa；
    S2，取经S1步软化的胡桃，超临界流体萃取，分离得到胡桃萃取液与胡桃残渣；所述超临界流体萃取的流体为二氧化碳，萃取温度为30～40℃；
    S3，取S2步得到的胡桃残渣，将其粉碎研磨；
    S4，取S2步得到的胡桃萃取液，将其喷雾干燥，喷雾干燥的过程中加入S3步得到的粉碎研磨后的胡桃残渣，得到干燥的胡桃萃取物；所述喷雾干燥的温度为40～80℃，干燥时间为3～60S；所述胡桃萃取物的颗粒度为10～60μm；
    S5，取1份S4步得到的胡桃萃取物，填充至天然植物胶囊壳内，得到胡桃萃取物胶囊；
    所述香艾萃取物胶囊的制备包括：
    S1，取香艾筛选、清洗，将干净的香艾的全草通过减压冷浸法软化；所述减压冷浸法的温度为10～40℃，压强为-0.1～-0.09MPa；
    S2，取经S1步软化的香艾，超临界流体萃取，分离得到香艾萃取液与香艾残渣；所述超临界流体萃取的流体为二氧化碳，萃取温度为30～40℃；
    S3，取S2步得到的香艾残渣，将其粉碎研磨；
    S4，取S2步得到的香艾萃取液，将其喷雾干燥，喷雾干燥的过程中加入S3步得到的粉碎研磨后的香艾残渣，得到干燥的香艾萃取物；所述喷雾干燥的温度为40～80℃，干燥时间为3～60S；所述香艾萃取物的颗粒度为10～60μm；
    S5，取54份S4步得到的香艾萃取物，填充至天然植物胶囊壳内，得到香艾萃取物胶囊；
    所述西洋参萃取物胶囊的制备包括：
    S1，取西洋参筛选、清洗，将干净的西洋参的参体通过减压冷浸法软化；所述减压冷浸法的温度为10～40℃，压强为-0.1～-0.09MPa；
    S2，取经S1步软化的西洋参，超临界流体萃取，分离得到西洋参萃取液与西洋参残渣；所述超临界流体萃取的流体为二氧化碳，萃取温度为30～40℃；
    S3，取S2步得到的西洋参残渣，将其粉碎研磨；
    S4，取S2步得到的西洋参萃取液，将其喷雾干燥，喷雾干燥的过程中加入S3 步得到的粉碎研磨后的西洋参残渣，得到干燥的西洋参萃取物；所述喷雾干燥的温度为40～80℃，干燥时间为3～60S；所述西洋参萃取物的颗粒度为10～60μm；
    S5，取1～5份S4步得到的西洋参萃取物，填充至天然植物胶囊壳内，得到西洋参萃取物胶囊；
    所述石斛萃取物胶囊的制备包括：
    S1，取石斛筛选、清洗，将干净的石斛茎叶通过减压冷浸法软化；所述减压冷浸法的温度为10～40℃，压强为-0.1～-0.09MPa；
    S2，取经S1步软化的石斛，超临界流体萃取，分离得到石斛萃取液与石斛残渣；所述超临界流体萃取的流体为二氧化碳，萃取温度为30～40℃；
    S3，取S2步得到的石斛残渣，将其粉碎研磨；
    S4，取S2步得到的石斛萃取液，将其喷雾干燥，喷雾干燥的过程中加入S3步得到的粉碎研磨后的石斛残渣，得到干燥的石斛萃取物；所述喷雾干燥的温度为40～80℃，干燥时间为3～60S；所述石斛萃取物的颗粒度为10～60μm；
    S5，取1～10份S4步得到的石斛萃取物，填充至天然植物胶囊壳内，得到石斛萃取物胶囊；
    所述胡桃为黑胡桃，香艾为中亚苦蒿，西洋参为半野生西洋参，石斛为野生石斛。
  8. 一种如权利要求1所述的抗红斑狼疮联合免疫用药物用以制备预防或***的药物的用途。
  9. 一种如权利要求5-7任一项所述的制备方法制备得到的抗红斑狼疮联合免疫用药物用以制备预防或***的药物的用途。
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Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20030068358A1 (en) * 2001-10-01 2003-04-10 Bogar Ag Preparation for oral administration
CN105169105A (zh) * 2015-10-13 2015-12-23 宁波御坊堂生物科技有限公司 具有抗肿瘤、滋阴养胃、增强免疫力功能的中药制剂及其制备方法
CN105770474A (zh) * 2016-04-07 2016-07-20 北京路德凯奇商贸有限责任公司 一种抗红斑狼疮联合免疫用药物及其制备方法、用途

Family Cites Families (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101757408A (zh) * 2009-12-16 2010-06-30 孙绍彬 一种***的药物及其制备方法
CN104322831A (zh) * 2014-10-20 2015-02-04 安徽益方堂生物工程有限公司 一种玫瑰菊花茶及其制备方法
CN105213850A (zh) * 2015-10-30 2016-01-06 姜秀芹 一种养阴润燥的营养品

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20030068358A1 (en) * 2001-10-01 2003-04-10 Bogar Ag Preparation for oral administration
CN105169105A (zh) * 2015-10-13 2015-12-23 宁波御坊堂生物科技有限公司 具有抗肿瘤、滋阴养胃、增强免疫力功能的中药制剂及其制备方法
CN105770474A (zh) * 2016-04-07 2016-07-20 北京路德凯奇商贸有限责任公司 一种抗红斑狼疮联合免疫用药物及其制备方法、用途

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
LI, QING: "Research and Application of Artemisia Annua", CHINA MEDICAL HERALD, vol. 5, no. 36, 31 December 2008 (2008-12-31), pages 25 - 26, XP009512791, ISSN: 1673-7210 *
LI, WEIYANG ET AL.: "Syndrome Differentiation Depending Treatment of Chinese Medicine of Systemic Lupus Erythematosus", SHAANXI JOURNAL OF TRADITIONAL CHINESE MEDICINE, vol. 23, no. 11, 30 November 2002 (2002-11-30), pages 1056 and 1057, XP009511814, ISSN: 1002-168X *
See also references of EP3441078A4 *
YANG, MINGJING ET AL.: "Effects of Dendrobium Candicum Capsules On Immune Responses In Mouse", JIANGSU JOURNAL OF PREVENTIVE MEDICINE, vol. 19, no. 04, 31 December 2008 (2008-12-31), pages 11 - 13, XP009511823 *

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