CN110974942A - Pharmaceutical composition, pharmaceutical bag, human-derived composite growth factor composite preparation, and preparation method and application thereof - Google Patents

Pharmaceutical composition, pharmaceutical bag, human-derived composite growth factor composite preparation, and preparation method and application thereof Download PDF

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CN110974942A
CN110974942A CN201911353687.6A CN201911353687A CN110974942A CN 110974942 A CN110974942 A CN 110974942A CN 201911353687 A CN201911353687 A CN 201911353687A CN 110974942 A CN110974942 A CN 110974942A
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growth factor
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vitamin
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CN110974942B (en
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张智勇
杨逸禧
周永林
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Guangdong Ruicheng Medical Technology Co ltd
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Third Affiliated Hospital of Guangzhou Medical University
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/18—Growth factors; Growth regulators
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    • A61K31/33—Heterocyclic compounds
    • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/35—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom
    • A61K31/352—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom condensed with carbocyclic rings, e.g. methantheline 
    • A61K31/353—3,4-Dihydrobenzopyrans, e.g. chroman, catechin
    • A61K31/355—Tocopherols, e.g. vitamin E
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
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    • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/365—Lactones
    • A61K31/375—Ascorbic acid, i.e. vitamin C; Salts thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/40—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil
    • A61K31/403—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil condensed with carbocyclic rings, e.g. carbazole
    • A61K31/404—Indoles, e.g. pindolol
    • A61K31/4045—Indole-alkylamines; Amides thereof, e.g. serotonin, melatonin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A61P17/02—Drugs for dermatological disorders for treating wounds, ulcers, burns, scars, keloids, or the like
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
    • A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

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Abstract

The invention discloses a pharmaceutical composition, a pharmaceutical bag, a human-derived composite growth factor composite preparation, a preparation method and an application thereof. The raw materials of the pharmaceutical composition comprise human-derived composite growth factors, vitamin C, vitamin E and melatonin after solvent removal, wherein the mass ratio of the vitamin C to the vitamin E to the melatonin is (1.76-8.8): (118.2-590.94):(2.32-11.61). The synergistic effect is generated by combining the human-derived composite growth factor, the vitamin C, the vitamin E and the melatonin, so that the inflammation level is reduced, the fibroblast proliferation and recovery are promoted in a diabetes environment, the formation of new blood vessels is promoted, and the composition can repair the diabetic wound.

Description

Pharmaceutical composition, pharmaceutical bag, human-derived composite growth factor composite preparation, and preparation method and application thereof
Technical Field
The invention relates to the technical field of pharmacy, in particular to a pharmaceutical composition, a drug bag, a human-derived composite growth factor composite preparation, a preparation method and an application thereof.
Background
The human-derived composite growth factor is an extract which is self-derived and rich in various growth factors, the sources of the extract comprise bone marrow, fat, blood platelets and the like, and the human-derived composite growth factor is currently applied to bone tissue repair. Platelet-derived human-derived composite growth factor-Platelet Rich Plasma (PRP) is widely used. Platelets, one of the major components in mammalian blood, are small, biologically active cytoplasm that is cleaved from the cytoplasm of the mature bone marrow megakaryocytes. Platelet-rich plasma (PRP) is a Platelet concentrate extracted from autologous blood by centrifugation, and has high concentrations of platelets, leukocytes, and fibrin as main components. Platelet Rich Plasma (PRP) therapy is an autologous regenerative repair therapy, mainly used for tissue wound repair. Such as fracture, bone defect, osteomyelitis, joint cartilage tendon ligament injury, and wound surface with difficulty in healing, such as pressure sore, burn, scald, and skin soft tissue defect. The technology adopts the self-derived platelet-rich plasma (PRP) for treatment, thus fundamentally avoiding the concern that the immunological rejection, the spread of diseases and the possibility of changing the human genetic structure of a heterogeneous recombinant gene product caused by an exogenous growth factor. The platelet-rich plasma (PRP) treatment technology has been developed in Europe, America and other countries for decades, and multi-center clinical observation is developed in China, which proves that the PRP treatment technology is safe and effective and can solve the problem of clinical tissue repair. However, regardless of the method used and despite technological improvements, Platelet-rich plasma (PRP) currently has a number of disadvantages when applied, in particular: 1. the growth factor content is low. 2. Blood must be collected during each treatment, and the patient can not use the blood by himself when leaving the hospital. 3. The storage time is short. The above disadvantages are due to the following reasons: 1. PRP is liquid, and the storage time is short; 2. PRP requires activation to release the most growth factors. Therefore, the preparation method of PRP is upgraded to form human-derived composite growth factor (ARF). However, the formed human-derived compound growth factor still has poor treatment effect on diabetic wounds.
In view of this, the invention is particularly proposed.
Disclosure of Invention
The invention aims to provide a pharmaceutical composition, a pharmaceutical bag, a human-derived composite growth factor composite preparation, a preparation method and application thereof.
The invention is realized by the following steps:
in a first aspect, the embodiments of the present invention provide a pharmaceutical composition, which comprises raw materials of human-derived complex growth factor, vitamin C, vitamin E and melatonin after solvent removal, wherein the mass ratio of the vitamin C to the vitamin E to the melatonin is (1.76-8.8): (118.2-590.94):(2.32-11.61).
In an alternative embodiment, the mass ratio of vitamin C, vitamin E and melatonin is (1.76-5.28): (236.38-472.75):(4.65-9.3).
In an alternative embodiment, the human-derived composite growth factor after removal of the solvent is a lyophilized human-derived composite growth factor;
preferably, the human-derived complex growth factor source includes at least one of blood, bone marrow, cord blood, and fat.
In a second aspect, embodiments of the present invention provide a pharmaceutical pack comprising a first component and a second component, wherein when the human-derived composite growth factor is a water-soluble human-derived composite growth factor, the first component comprises vitamin C and the human-derived composite growth factor after removal of a solvent, and the second component comprises vitamin E and melatonin;
when the human-derived composite growth factor is soluble in an organic solvent, the first component comprises vitamin C, and the second component comprises the human-derived composite growth factor after the solvent is removed, vitamin E and melatonin;
the mass ratio of the vitamin C, the vitamin E and the melatonin is (1.76-8.8): (118.2-590.94) (2.32-11.61), preferably (1.76-5.28): (236.38-472.75): (4.65-9.3); the first component and the second component are mixed while using the pharmaceutical pack.
In a third aspect, the present invention provides a human-derived complex growth factor complex formulation, which comprises the pharmaceutical composition according to any one of the preceding embodiments or the pharmaceutical pack according to the preceding embodiments.
In alternative embodiments, the concentration of vitamin C in the human-derived complex growth factor complex formulation is 10-50 μmol/L, preferably 10-30 μmol/L, more preferably 20 μmol/L;
preferably, the concentration of the vitamin E in the human-derived composite growth factor composite preparation is 0.25-1.25mmol/L, preferably 0.5-1mmol/L, and more preferably 1 mmol/L;
preferably, the concentration of the melatonin in the human-derived composite growth factor composite preparation is 10-50 mu mol/L, preferably 20-40 mu mol/L, and more preferably 20 mu mol/L;
the concentration of the human-derived composite growth factor in the human-derived composite growth factor preparation is consistent with that of the human-derived composite growth factor before the solvent is removed.
In a fourth aspect, the present invention provides a method for preparing a human-derived complex growth factor complex preparation according to the foregoing embodiments, including preparing the human-derived complex growth factor complex preparation from the pharmaceutical composition according to any one of the foregoing embodiments or the pharmaceutical package according to the foregoing embodiments.
In an alternative embodiment, when the pharmaceutical composition is prepared as a human-derived complex growth factor complex preparation, the method comprises mixing the pharmaceutical composition with a complex solvent to form the human-derived complex growth factor complex preparation.
In an alternative embodiment, when the human-derived composite growth factor is a water-soluble human-derived composite growth factor, the composite solvent is a mixed solution formed by mixing a first solvent capable of dissolving the human-derived composite growth factor and vitamin C, respectively, and a second solvent capable of dissolving vitamin E and melatonin, respectively, and the first solvent and the second solvent are mutually soluble;
when the human-derived composite growth factor is a human-derived composite growth factor capable of being dissolved in an organic solvent, the composite solvent is a mixed solution formed by mixing a first solvent capable of dissolving the biotin C and a second solvent capable of respectively dissolving the vitamin E, the human-derived composite growth factor and the melatonin, and the first solvent and the second solvent are mutually soluble;
preferably, the first solvent is a polar solvent, preferably water;
preferably, the second solvent is an alcoholic solvent, preferably a monohydric alcohol solvent, more preferably ethanol;
preferably, the volume ratio of the first solvent to the second solvent is 4-10:1, preferably 8: 1.
In an alternative embodiment, preparing the pharmaceutical pack as a complex human-derived growth factor complex formulation comprises dissolving the first component to form a first solution;
dissolving the second component to form a second solution;
then mixing the first solution and the second solution to form the human-derived composite growth factor composite preparation;
preferably, the preparation of the first solution comprises: mixing the first component with a first solvent such that the first component is dissolved;
preferably, the first solvent is a polar solvent, more preferably water;
the preparation of the second solution comprises: mixing a second component with a second solvent such that the second component is dissolved and the second solvent is miscible with the first solvent;
preferably, the second solvent is an alcoholic solvent, preferably a monohydric alcohol solvent, more preferably ethanol;
preferably, the volume ratio of the first solvent to the second solvent is 4-10:1, preferably 8: 1.
In a fifth aspect, the embodiments of the present invention provide a pharmaceutical composition according to any one of the preceding embodiments, a pharmaceutical pack according to any one of the preceding embodiments, a human-derived composite growth factor complex preparation according to any one of the preceding embodiments, and an application of the human-derived composite growth factor complex preparation prepared by the preparation method according to any one of the preceding embodiments in preparing a medicament for treating a diabetic wound.
The invention has the following beneficial effects: the invention combines human-derived composite growth factors, vitamin C, vitamin E and melatonin and limits the proportion of the growth factors, the vitamin C, the vitamin E and the melatonin, so that the substances generate synergistic action to reduce the inflammation level, promote the proliferation and recovery of fibroblasts in a diabetes environment, promote the formation of new blood vessels and further repair the wound surface of diabetes by the composition.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings needed to be used in the embodiments will be briefly described below, it should be understood that the following drawings only illustrate some embodiments of the present invention and therefore should not be considered as limiting the scope, and for those skilled in the art, other related drawings can be obtained according to the drawings without inventive efforts.
FIG. 1 is a graph showing the results of detection provided in Experimental example 1 of the present invention;
FIG. 2 is a diagram showing the results of the test provided in Experimental example 2 of the present invention.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products available commercially.
The features and properties of the present invention are described in further detail below with reference to examples.
The embodiment of the invention provides a pharmaceutical composition, which comprises the following raw materials of human-derived composite growth factors, vitamin C, vitamin E and melatonin after solvent removal, wherein the mass ratio of the vitamin C to the vitamin E to the melatonin is (1.76-8.8): (118.2-590.94) (2.32-11.61), preferably (1.76-5.28): (236.38-472.75):(4.65-9.3). The vitamin C, the vitamin E, the human-derived composite growth factor and the melatonin are combined, so that the inflammation level can be reduced, the fibroblast proliferation and recovery are promoted in a diabetes environment, the formation of new blood vessels is promoted, and the repair of the medicine to the diabetic wound surface is promoted. And the specific mass ratio among the limited vitamin C, the limited vitamin E and the limited melatonin further ensures the combined effect and the repair effect on the diabetic wound. Meanwhile, the solvent of the human-derived composite growth factor is removed, so that the pharmaceutical composition is more favorably stored, the pharmaceutical composition is prevented from going bad, and the treatment effect of the pharmaceutical composition is ensured.
Specifically, vitamin C and vitamin E are both strong antioxidants, cells in an oxidation environment can be protected more effectively by combining the vitamin C and the vitamin E, melatonin is a free radical scavenger, the vitamin C and the melatonin can generate a synergistic effect, free radicals in a body can be effectively scavenged, damage caused by the free radicals is reduced, inflammation is further inhibited after the vitamin C, the vitamin E and the melatonin are combined, and tissue regeneration is promoted.
Further, the human-derived composite growth factor after the solvent is removed is a lyophilized human-derived composite growth factor; preferably, the human-derived complex growth factor source comprises at least one of blood, cord blood, bone marrow, and fat. Namely, the platelet-rich plasma, fat or bone marrow is subjected to freeze-drying treatment, the solvent in the liquid is removed, and the solute is remained, so as to obtain the freeze-dried human-derived composite growth factor. The human-derived composite growth factor without the solvent is not only favorable for preservation, but also ensures the high content of the growth factor after redissolution, and is more favorable for repairing the wound surface.
It should be noted that the method for removing the solvent is not limited to lyophilization, and any treatment method that removes the solvent without destroying the activity of the human-derived complex growth factor after solvent removal-reconstitution in the prior art may be employed.
The pharmaceutical composition is formed by directly mixing the raw materials.
The embodiment of the invention also provides a medicine package which comprises a first component and a second component, wherein the first component and the second component are used as the same component with the raw materials with the same solubility according to the solubility of the growth factors, and the rest other raw materials are used as the other component, so that the subsequent redissolution and the storage of the medicine are facilitated. For example, when the human-derived composite growth factor is a water-soluble human-derived composite growth factor, the first component includes vitamin C and the human-derived composite growth factor after removal of the solvent, and the second component includes vitamin E and melatonin; when the human-derived composite growth factor is soluble in an organic solvent, the first component comprises vitamin C, and the second component comprises the human-derived composite growth factor after the solvent is removed, vitamin E and melatonin; even the human-derived composite growth factor, the vitamin C, the vitamin E and the melatonin which are subjected to solvent removal can be taken as single components, and the components are mixed when being redissolved for use.
The mass ratio of the human-derived composite growth factor to the vitamin C to the vitamin E to the melatonin is (1.7612-8.8): (118.2-590.94) (2.32-11.61), preferably (1.76-5.28): (236.38-472.75): (4.65-9.3); more preferably 3.52: 472.75: 4.65; the first component and the second component are mixed while using the pharmaceutical pack.
The embodiment of the invention also provides a human-derived composite growth factor compound preparation, which comprises the pharmaceutical composition of any one of the previous embodiments or the pharmaceutical package of the previous embodiments.
Further, the concentration of the vitamin C in the human-derived composite growth factor composite preparation is 10-50 mu mol/L, preferably 10-30 mu mol/L, and more preferably 20 mu mol/L;
preferably, the concentration of the vitamin E in the human-derived composite growth factor composite preparation is 0.25-1.25mmol/L, preferably 0.5-1mmol/L, and more preferably 1 mmol/L;
preferably, the concentration of the melatonin in the human-derived composite growth factor composite preparation is 10-50 mu mol/L, preferably 20-40 mu mol/L, more preferably 20 mu mol/L,
the concentration of the human-derived composite growth factor in the human-derived composite growth factor composite preparation is consistent with that of the human-derived composite growth factor before the solvent is removed, so that the human-derived composite growth factor can be ensured to exert the efficacy.
The human-derived composite growth factor preparation is a gel or a suspension or other dosage forms formed after the pharmaceutical composition or the drug bag is redissolved, and the volume, the concentration and the like of the human-derived composite growth factor preparation after redissolution are the same as those of the human-derived composite growth factor before freeze-drying or before solvent removal, namely are the same as those of the human-derived composite growth factor without freeze-drying or solvent removal, so that the quantity of the human-derived composite growth factor in the human-derived composite growth factor preparation can be ensured to be unchanged, and then the sufficient growth factor is ensured.
The concentration of vitamin C, vitamin E and melatonin in the human-derived composite growth factor preparation is limited, so that the synergistic effect of the vitamin C, the vitamin E and the melatonin in the human-derived composite growth factor preparation can be ensured, the inflammation is reduced, the formation of new blood vessels is accelerated, and the repair of diabetic wounds is facilitated.
The embodiment of the invention also provides a preparation method of the human-derived composite growth factor preparation, which comprises the step of preparing the pharmaceutical composition or the medicine into the human-derived composite growth factor preparation.
The difference between the human-derived composite growth factor preparation prepared by mixing the pharmaceutical composition and the solvent directly, and the human-derived composite growth factor preparation prepared by using the pharmaceutical pack, wherein the first component and the second component are dissolved respectively and then mixed, but the type and the amount of the specific solvent formed finally are the same.
Specifically, the following:
when the pharmaceutical composition is prepared into the human-derived composite growth factor composite preparation, the pharmaceutical composition and a composite solvent are mixed to form the human-derived composite growth factor composite preparation.
In an alternative embodiment, when the human-derived composite growth factor is a water-soluble human-derived composite growth factor, the composite solvent is a mixed solution formed by mixing a first solvent capable of dissolving the human-derived composite growth factor and vitamin C, respectively, and a second solvent capable of dissolving vitamin E and melatonin, respectively, and the first solvent and the second solvent are mutually soluble;
when the human-derived composite growth factor is a human-derived composite growth factor capable of being dissolved in an organic solvent, the composite solvent is a mixed solution formed by mixing a first solvent capable of dissolving the biotin C and a second solvent capable of respectively dissolving the vitamin E, the human-derived composite growth factor and the melatonin, and the first solvent and the second solvent are mutually soluble;
preferably, the first solvent is a polar solvent, preferably water;
preferably, the second solvent is an alcoholic solvent, preferably a monohydric alcohol solvent, more preferably ethanol;
preferably, the volume ratio of the first solvent to the second solvent is 4-10:1, preferably 8: 1.
Firstly, preparing a composite solvent, namely mixing a first solvent and a second solvent to form a mixed solvent capable of simultaneously dissolving the human-derived composite growth factor, the vitamin C, the vitamin E and the melatonin, then mixing the composite solvent and the pharmaceutical composition to dissolve the pharmaceutical composition, and obtaining the human-derived composite growth factor preparation with corresponding concentration. The volume ratio of the first solvent to the second solvent is controlled, so that the composite solvent is more favorable for dissolving each substance, and the treatment effect of the composite growth factor preparation derived from people is further ensured.
Or when the drug bag is prepared into a human-derived composite growth factor composite preparation, the method comprises the steps of dissolving the first component to form a first solution;
dissolving the second component to form a second solution;
then mixing the first solution and the second solution to form the human-derived composite growth factor composite preparation;
preferably, the preparation of the first solution comprises: mixing the first component with a first solvent such that the first component is dissolved;
preferably, the first solvent is a polar solvent, more preferably water;
the preparation of the second solution comprises: mixing a second component with a second solvent such that the second component is dissolved and the second solvent is miscible with the first solvent;
preferably, the second solvent is an alcoholic solvent, preferably a monohydric alcohol solvent, more preferably ethanol;
preferably, the volume ratio of the first solvent to the second solvent is 4-10:1, preferably 8: 1.
When the medicine bag is used, the first component and the second component are dissolved respectively, and then the dissolved solutions are mixed, so that the human-derived composite growth factor preparation can be obtained. The preparation method can further ensure the treatment effect of the human-derived composite growth factor composite preparation on the diabetic wound.
The first solvent may be selected to dissolve the first component, and the second solvent may be selected to dissolve the second component and be miscible with the first solvent, regardless of the presence or absence of the human-derived complex growth factor in the first component.
Example 1
The embodiment provides a pharmaceutical composition, which comprises 0.3 g of human-derived composite growth factor powder after solvent removal (in the application, the human-derived composite growth factor powder after solvent removal is platelet-enriched plasma freeze-dried powder), 3.52 micrograms of vitamin C, 472.75 micrograms of vitamin E and 4.65 micrograms of melatonin.
This example provides a human-derived composite growth factor preparation, wherein the concentration of vitamin C in the human-derived composite growth factor preparation is 20 μmol/L, the concentration of vitamin E is 1mmol/L, the concentration of melatonin is 20 μmol/L, and the number of platelets is 1 × 1010And (2) per liter.
The embodiment provides a preparation method of a human-derived composite growth factor composite preparation, which comprises the following steps:
mixing water and ethanol according to a volume ratio of 8:1 to prepare a composite solvent;
the composite solvent is then mixed with the pharmaceutical composition and stirred to dissolve the pharmaceutical composition.
Example 2
The embodiment provides a pharmaceutical composition, which comprises 0.3 g of human-derived composite growth factor (platelet-rich plasma freeze-dried powder), 8.8 micrograms of vitamin C, 472.75 micrograms of vitamin E and 2.32 micrograms of melatonin.
This example provides a human-derived composite growth factor preparation, wherein the concentration of vitamin C in the human-derived composite growth factor preparation is 50 μmol/L, the concentration of vitamin E is 1.25mmol/L, the concentration of melatonin is 10 μmol/L, and the number of platelets is 1 × 1010And (2) per liter.
The embodiment provides a preparation method of a human-derived composite growth factor composite preparation, which comprises the following steps:
mixing water and ethanol according to a volume ratio of 8:1 to prepare a composite solvent;
the composite solvent is then mixed with the pharmaceutical composition and stirred to dissolve the pharmaceutical composition.
Example 3
The embodiment provides a pharmaceutical composition, which comprises 0.3 g of human-derived composite growth factor (platelet-rich plasma freeze-dried powder), 1.76 micrograms of vitamin C, 118.2 micrograms of vitamin E and 11.61 micrograms of melatonin.
This example provides a human-derived composite growth factor preparation, wherein the concentration of vitamin C in the human-derived composite growth factor preparation is 10 μmol/L, the concentration of vitamin E is 0.25mmol/L, the concentration of melatonin is 50 μmol/L, and the number of platelets is 1.6 × 1010And (2) per liter.
The embodiment provides a preparation method of a human-derived composite growth factor composite preparation, which comprises the following steps:
mixing water and ethanol according to a volume ratio of 8:1 to prepare a composite solvent;
the composite solvent is then mixed with the pharmaceutical composition and stirred to dissolve the pharmaceutical composition.
Example 4
The embodiment provides a medicine package which comprises a first component and a second component, wherein the first component comprises 0.3 g of human-derived composite growth factor (platelet-rich plasma freeze-dried powder) and 3.52 micrograms of vitamin C after solvent removal, and the second component comprises 472.75 micrograms of vitamin E and 4.65 micrograms of melatonin.
This example provides a human-derived composite growth factor preparation, wherein the concentration of vitamin C in the human-derived composite growth factor preparation is 20 μmol/L, the concentration of vitamin E is 1mmol/L, the concentration of melatonin is 20 μmol/L, and the number of platelets is 0.8 × 103And (2) per liter.
The embodiment provides a preparation method of a human-derived composite growth factor composite preparation, which comprises the following steps:
mixing and stirring water and the first component to dissolve the first component to form a first solution;
mixing and stirring ethanol and a second component to dissolve the second component to form a second solution, wherein the volume ratio of the water to the ethanol is 8: 1;
and mixing the first solution and the second solution to form the human-derived composite growth factor compound preparation.
Example 5
The embodiment provides a medicine package which comprises a first component and a second component, wherein the first component comprises 0.3 g of human-derived composite growth factor (platelet-rich plasma freeze-dried powder) and 5.28 micrograms of vitamin C after solvent removal, and the second component comprises 236.38 micrograms of vitamin E and 9.3 micrograms of melatonin.
This example provides a human-derived composite growth factor preparation, wherein the concentration of vitamin C in the human-derived composite growth factor preparation is 30 μmol/L, the concentration of vitamin E is 0.5mmol/L, the concentration of melatonin is 40 μmol/L, and the number of platelets is 2.0 × 1010And (2) per liter.
The embodiment provides a preparation method of a human-derived composite growth factor composite preparation, which comprises the following steps:
mixing and stirring water and the first component to dissolve the first component to form a first solution;
mixing and stirring ethanol and a second component to dissolve the second component to form a second solution, wherein the volume ratio of the water to the ethanol is 4: 1;
and mixing the first solution and the second solution to form the human-derived composite growth factor compound preparation.
Example 6
The embodiment provides a medicine package which comprises a first component and a second component, wherein the first component comprises 0.3 g of human-derived composite growth factor (platelet-rich plasma freeze-dried powder) and 7.32 micrograms of vitamin C after solvent removal, and the second component comprises 590.94 micrograms of vitamin E and 8.4 micrograms of melatonin.
This example provides a human-derived composite growth factor preparation, wherein the concentration of vitamin C in the human-derived composite growth factor preparation is 25 μmol/L, the concentration of vitamin E is 0.75mmol/L, the concentration of melatonin is 35 μmol/L, and the number of platelets is 0.6 × 1010And (2) per liter.
The embodiment provides a preparation method of a human-derived composite growth factor composite preparation, which comprises the following steps:
mixing and stirring water and the first component to dissolve the first component to form a first solution;
mixing and stirring ethanol and a second component to dissolve the second component to form a second solution, wherein the volume ratio of the water to the ethanol is 10: 1;
and mixing the first solution and the second solution to form the human-derived composite growth factor compound preparation.
Experimental example 1
The operation method comprises the following steps:
(1) fibroblasts (fibroplasts) were plated in 96-well plates, approximately 6-full, overnight, and the cells were divided into 7 groups (Control, AGE group, VC + E group, melatonin group, VC + melatonin group, VE + melatonin group, and example 1 group), and each group had 5 wells.
Note: AGEs are advanced glycation end products (AGEs) that can mimic the diabetic environment in cell culture.
(2) The next day, the medium in the wells was aspirated and different drugs (concentration values of drug in medium) were added.
Control group: adding new cell culture medium;
AGE group: new cell culture medium, AGEs (concentration 400 μ M);
VitC + E group: new cell culture media, AGEs (400. mu.M concentration), VC (50. mu. mol/L concentration and VE (1.25 mmol/L concentration);
VC + melatonin group novel cell culture Medium, AGEs (concentration 400. mu.M), VC (concentration 50. mu. mol/L), melatonin (concentration 50. mu. mol/L)
VE + melatonin group: new cell culture media, AGEs (400. mu.M concentration), VE (1.25 mmol/L concentration), melatonin (50. mu. mol/L concentration)
Melatonin group: new cell culture medium, AGEs (concentration 400 μ M), melatonin (concentration 50 μmol/L);
example 1 group: novel cell culture media, AGEs (400. mu.M concentration), Vit C (20. mu. mol/L concentration), Vit (1 mmol/L concentration), melatonin (20. mu. mol/L concentration);
(3) after 24h, the cell proliferation is detected by using a CCK8 method, and the detection result is shown in figure 1.
The concentrations in the experimental examples represent the concentrations of the respective substances in the medium.
As can be seen from fig. 1, the cellular proliferation of the VC + E group was statistically different from that of the AGE group in the melatonin group, and the cellular proliferation of the example 1 group was also statistically different from that of the VC + E group in the melatonin group. It can therefore be shown that when VitC, E are combined with melatonin, fibroblast proliferation recovery is better in a diabetic setting than when vitamin or melatonin alone is used.
Experimental example 2
The operation method comprises the following steps:
(1) matrigel was spread in 24-well plates, endothelial cells were spread overnight, about 6-full, overnight, cells were divided into 7 groups (Control, AGE group, VC + E group, melatonin group, VC + melatonin group, VE + melatonin group and example 1 group), 3 wells per group.
Note: AGEs are advanced glycation end products (AGEs) that can mimic the diabetic environment in cell culture.
(2) The next day, the medium in the wells was aspirated and different drugs (concentration values of drug in medium) were added.
Control group: adding new cell culture medium;
AGE group: new cell culture medium, AGEs (concentration 400 μ M);
VitC + E group: new cell culture media, AGEs (400. mu.M concentration), VC (50. mu. mol/L concentration and VE (1.25 mmol/L concentration);
VC + melatonin group novel cell culture Medium, AGEs (concentration 400. mu.M), VC (concentration 50. mu. mol/L), melatonin (concentration 50. mu. mol/L)
VE + melatonin group: new cell culture media, AGEs (400. mu.M concentration), VE (1.25 mmol/L concentration), melatonin (50. mu. mol/L concentration)
Melatonin group: new cell culture medium, AGEs (concentration 400 μ M), melatonin (concentration 50 μmol/L);
example 1 group: novel cell culture media, AGEs (400. mu.M concentration), Vit C (20. mu. mol/L concentration), Vit (1 mmol/L concentration), melatonin (20. mu. mol/L concentration);
(3) after 24h, the vascularization condition of the cells is detected by using a microscope statistical method, the total area of the blood vessels is counted by using ImageJ software, and the detection result is shown in figure 2. As can be seen from fig. 2, when the human-derived composite growth factor is used in combination with Vit C, Vit E, and melatonin, the endothelial cell vascularization effect is optimal in the diabetic environment, so that sufficient neovascularization can be ensured to promote the healing of the wound surface when the diabetic wound heals early.
The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention, and various modifications and changes may be made by those skilled in the art. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims (10)

1. The pharmaceutical composition is characterized in that raw materials of the pharmaceutical composition comprise human-derived compound growth factors, vitamin C, vitamin E and melatonin after solvent removal, wherein the mass ratio of the vitamin C to the vitamin E to the melatonin is (1.76-8.8): (118.2-590.94):(2.32-11.61).
2. The pharmaceutical composition according to claim 1, wherein the mass ratio of vitamin C, vitamin E and melatonin is (1.76-5.28): (236.38-472.75):(4.65-9.3).
3. The pharmaceutical composition of claim 1, wherein the human-derived composite growth factor after solvent removal is a lyophilized human-derived composite growth factor;
preferably, the source of the human-derived complex growth factor includes at least one of blood, bone marrow, umbilical cord blood, fat, and the like.
4. A pharmaceutical pack characterized by comprising a first component and a second component, wherein when the human-derived composite growth factor is a water-soluble human-derived composite growth factor, the first component comprises vitamin C and the human-derived composite growth factor after removal of a solvent, and the second component comprises vitamin E and melatonin;
when the human-derived composite growth factor is soluble in an organic solvent, the first component comprises vitamin C, and the second component comprises the human-derived composite growth factor after the solvent is removed, vitamin E and melatonin;
the mass ratio of the vitamin C, the vitamin E and the melatonin in the first component and the second component is (1.7612-8.8): (118.2-590.94) (2.32-11.61), preferably (1.76-5.28): (236.38-472.75): (4.65-9.3); the first component and the second component are mixed while using the pharmaceutical pack.
5. A complex formulation of human-derived complex growth factors, comprising the pharmaceutical composition of any one of claims 1 to 3 or the pharmaceutical pack of claim 4.
6. The complex human-derived growth factor preparation according to claim 5, wherein the concentration of vitamin C in the complex human-derived growth factor preparation is 10 to 50 μmol/L, preferably 10 to 30 μmol/L;
preferably, the concentration of the vitamin E in the human-derived composite growth factor composite preparation is 0.25-1.25mmol/L, preferably 0.5-1 mmol/L;
preferably, the concentration of the melatonin in the human-derived composite growth factor composite preparation is 10-50 mu mol/L, preferably 20-40 mu mol/L,
the concentration of the human-derived composite growth factor in the human-derived composite growth factor composite preparation is consistent with that of the human-derived composite growth factor before the solvent is removed;
preferably, when the human-derived composite growth factor is platelets, the amount of the human-derived composite growth factor composite preparation is 0.6-2.0x1010Per liter, preferably 0.8-1.6x1010And (2) per liter.
7. The method for preparing the human-derived complex growth factor complex preparation according to claim 5 or 6, comprising preparing the pharmaceutical composition according to any one of claims 1 to 3 or the pharmaceutical composition according to claim 4 into the human-derived complex growth factor complex preparation.
8. The preparation method according to claim 7, wherein the preparation of the pharmaceutical composition as the human-derived complex growth factor complex preparation comprises mixing the pharmaceutical composition with a complex solvent to form the human-derived complex growth factor complex preparation;
preferably, when the human-derived composite growth factor is a water-soluble human-derived composite growth factor, the composite solvent is a mixed solution formed by mixing a first solvent capable of dissolving the human-derived composite growth factor and vitamin C respectively and a second solvent capable of dissolving vitamin E and melatonin respectively, and the first solvent and the second solvent are mutually soluble;
when the human-derived composite growth factor is a human-derived composite growth factor capable of being dissolved in an organic solvent, the composite solvent is a mixed solution formed by mixing a first solvent capable of dissolving the biotin C and a second solvent capable of respectively dissolving the vitamin E, the human-derived composite growth factor and the melatonin, and the first solvent and the second solvent are mutually soluble;
preferably, the first solvent is a polar solvent, preferably water;
preferably, the second solvent is an alcoholic solvent, preferably a monohydric alcohol solvent, more preferably ethanol;
preferably, the volume ratio of the first solvent to the second solvent is 4-10:1, preferably 8: 1.
9. The method of claim 7, wherein the pharmaceutical pack is prepared as a complex human-derived growth factor complex formulation by dissolving the first component to form a first solution;
dissolving the second component to form a second solution;
then mixing the first solution and the second solution to form the human-derived composite growth factor composite preparation;
preferably, the preparation of the first solution comprises: mixing the first component with a first solvent such that the first component is dissolved;
preferably, the first solvent is a polar solvent, more preferably water;
the preparation of the second solution comprises: mixing a second component with a second solvent such that the second component is dissolved and the second solvent is miscible with the first solvent;
preferably, the second solvent is an alcoholic solvent, preferably a monohydric alcohol solvent, more preferably ethanol;
preferably, the volume ratio of the first solvent to the second solvent is 4-10:1, preferably 8: 1.
10. Use of the pharmaceutical composition according to any one of claims 1 to 3, the pharmaceutical pack according to claim 4, the human-derived complex growth factor complex preparation according to claim 5 or 6, and the human-derived complex growth factor complex preparation prepared by the preparation method according to any one of claims 7 to 9 in the preparation of a medicament for treating diabetic wounds.
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Citations (1)

* Cited by examiner, † Cited by third party
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CN102988964A (en) * 2012-11-02 2013-03-27 ***广州总医院 Compound growth factor as well as preparation method and application thereof

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102988964A (en) * 2012-11-02 2013-03-27 ***广州总医院 Compound growth factor as well as preparation method and application thereof

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Title
贾志刚: "抗氧化剂与创伤修复的关系", 《医学综述》 *

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