CN114831939A - Triamcinolone acetonide acetate composition spray and preparation method thereof - Google Patents

Triamcinolone acetonide acetate composition spray and preparation method thereof Download PDF

Info

Publication number
CN114831939A
CN114831939A CN202110131270.6A CN202110131270A CN114831939A CN 114831939 A CN114831939 A CN 114831939A CN 202110131270 A CN202110131270 A CN 202110131270A CN 114831939 A CN114831939 A CN 114831939A
Authority
CN
China
Prior art keywords
spray
dspe
peg2000
triamcinolone acetonide
sst
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Granted
Application number
CN202110131270.6A
Other languages
Chinese (zh)
Other versions
CN114831939B (en
Inventor
刘铠豪
刘庭福
姚志勇
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
NANJING XINGYIN PHARMACEUTICAL GROUP CO Ltd
Original Assignee
NANJING XINGYIN PHARMACEUTICAL GROUP CO Ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by NANJING XINGYIN PHARMACEUTICAL GROUP CO Ltd filed Critical NANJING XINGYIN PHARMACEUTICAL GROUP CO Ltd
Priority to CN202110131270.6A priority Critical patent/CN114831939B/en
Publication of CN114831939A publication Critical patent/CN114831939A/en
Application granted granted Critical
Publication of CN114831939B publication Critical patent/CN114831939B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • 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
    • A61K31/56—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids
    • A61K31/58—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids containing heterocyclic rings, e.g. danazol, stanozolol, pancuronium or digitogenin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/04—Peptides having up to 20 amino acids in a fully defined sequence; Derivatives thereof
    • A61K38/12—Cyclic peptides, e.g. bacitracins; Polymyxins; Gramicidins S, C; Tyrocidins A, B or C
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
    • A61K47/24—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing atoms other than carbon, hydrogen, oxygen, halogen, nitrogen or sulfur, e.g. cyclomethicone or phospholipids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
    • A61K47/28—Steroids, e.g. cholesterol, bile acids or glycyrrhetinic acid
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/56—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule
    • A61K47/59—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyureas or polyurethanes
    • A61K47/60—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyureas or polyurethanes the organic macromolecular compound being a polyoxyalkylene oligomer, polymer or dendrimer, e.g. PEG, PPG, PEO or polyglycerol
    • 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
    • A61K9/0012—Galenical forms characterised by the site of application
    • A61K9/0014—Skin, i.e. galenical aspects of topical compositions
    • 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
    • A61K9/10—Dispersions; Emulsions
    • A61K9/12—Aerosols; Foams
    • 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
    • A61K9/10—Dispersions; Emulsions
    • A61K9/127—Liposomes
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P19/00—Drugs for skeletal disorders
    • A61P19/02—Drugs for skeletal disorders for joint disorders, e.g. arthritis, arthrosis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P5/00—Drugs for disorders of the endocrine system
    • A61P5/38—Drugs for disorders of the endocrine system of the suprarenal hormones
    • A61P5/44—Glucocorticosteroids; Drugs increasing or potentiating the activity of glucocorticosteroids

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Medicinal Chemistry (AREA)
  • Epidemiology (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Dispersion Chemistry (AREA)
  • Oil, Petroleum & Natural Gas (AREA)
  • Immunology (AREA)
  • Organic Chemistry (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Dermatology (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Orthopedic Medicine & Surgery (AREA)
  • Rheumatology (AREA)
  • Physical Education & Sports Medicine (AREA)
  • Molecular Biology (AREA)
  • Biophysics (AREA)
  • Diabetes (AREA)
  • Endocrinology (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

The invention discloses a triamcinolone acetonide acetate composition spray and a preparation method thereof, belonging to the field of pharmaceutical preparations. A triamcinolone acetonide acetate composition spray comprises triamcinolone acetonide acetate, somatostatin modifier, DSPE-PEG2000, phospholipid, cholesterol, liposome membrane regulator and pharmaceutically acceptable carrier; wherein the spray comprises a carrier, the carrier comprises DSPE-PEG2000, phospholipid, cholesterol and liposome membrane regulator, the particle size range is 40-100 nm, and at least 80% of the carrier particle size is uniform. The invention combines triamcinolone acetonide acetate and somatostatin for administration, relieves the arthralgia and swelling from different mechanisms, has synergistic effect, and has quick response and lasting treatment effect. In addition, the prepared transfersome has small and uniform particle size and good transdermal effect.

Description

Triamcinolone acetonide acetate composition spray and preparation method thereof
Technical Field
The invention relates to the field of pharmaceutical preparations, in particular to a triamcinolone acetonide acetate composition spray and a preparation method thereof.
Background
Triamcinolone acetonide (triamcinolone acetonide), also called triamcinolone acetonide, triamcinolone acetonide and the like, is a middle-acting glucocorticoid, has similar action to triamcinolone and hydrocortisone, has the effects of resisting inflammation, resisting pruritus, contracting blood vessels and the like, has weak retention of sodium and water, and has stronger and lasting anti-inflammatory action and 20-30 times of efficacy compared with cortisone. Triamcinolone acetonide acetate is generally clinically divided into injection and ointment, and is suitable for various skin diseases, allergic rhinitis, arthralgia, bronchial asthma, scapulohumeral periarthritis, tenosynovitis, synovitis, acute sprain, rheumatoid arthritis and the like.
Somatostatin (SST), a cyclic peptide of 14 amino acids produced by the central nervous system, having the amino acid sequence: H-Ala 1 -Gly 2 -Cys 3 -Lys 4 -Asn 5 -Phe 6 -Phe 7 -Trp 8 -Lys 9 -Thr 10 -Phe 11 -Thr 12 -Ser 13 -Cys 14 -OH, a disulfide bond between Cys at positions 3 and 14. Somatostatin is a neurohormone with wider action, has the main effects of inhibiting the basal secretion of pituitary Growth Hormone (GH) and inhibiting GH secretory reaction caused by adenohypophysis to various stimuli, and is widely applied to severe upper gastrointestinal hemorrhage, acute pancreatitis and postoperative complications of pancreatic surgery. In 1988, Matucci and Marabini reported the treatment of rheumatoid arthritis by intraarticular injection of SST (Matucci-cerinic M, Marabini S. somatotitantegramtent for pain in rheumoid arthritis: adorable blue shows plant in knee infusion. Med Sci Res, 1988,16: 233-. Thereafter, SST injection has also been used to treat psoriatic arthritis, scapulohumeral periarthritis, osteoarthritis, and the like.
Arthritis (arthritis) generally refers to the occurrence of joints and their surrounding tissues in the human body, and there are several tens of inflammatory diseases caused by inflammation, infection, degeneration, trauma or other factors. Arthritis patients in China are more than 1 hundred million, and the number of the arthritis patients is increasing continuously. The clinical manifestations are red, swelling, heat, pain, dysfunction and joint deformity of joints, and severe patients cause joint disability and affect the life quality of patients. Clinically, 10 mg/mL or 40 mg/mL triamcinolone acetonide is usually injected into a bone joint to treat acute gouty arthritis or rheumatoid arthritis, and after repeated multiple injections of the triamcinolone acetonide closed needle, the following complications can occur: 1. degeneration or dissolution of the tendon, which results after multiple injections, is prone to spontaneous rupture and may also lead to atrophy of the tendon. 2. Degeneration of cartilage, such as thinning, abrasion and increased degeneration of cartilage, is easily caused. 3. Repeated injections into relatively thin areas of the skin result in a reduction in the skin's pigment levels and local skin whitening. 4. Repeated injections or injections tend to cause a decrease in local resistance and to produce bacterial infections.
Triamcinolone acetonide and SST are required to be injected for administration when treating arthritis, so that the toxic and side effects of the medicament are increased, patients are very painful, the medicament is inconvenient to use, and other complications are easy to generate after multiple injections. Therefore, the development of a safe and effective transdermal preparation with convenient application is significant.
The stratum corneum has a natural barrier to most molecules, especially macromolecules such as polypeptides. Transfersomes (TF), also known as deformed liposomes, flexible nanoliposomes or flexible liposomes, are self-aggregating vesicles, the main components of which are phospholipids and surfactants (such as sodium cholate, sodium deoxycholate and the like), and through the high deformability of the transfersomes, water and gradient existing in the skin are used as driving forces under the condition of non-closed administration, so that the self-aggregating vesicles can efficiently pass through the pores with the diameter being several times smaller than that of the self-aggregating vesicles. Water-soluble and fat-soluble macromolecules and micromolecules can smoothly pass through the skin by taking a carrier as a carrier, can target deep skin tissues to reach the internal circulation to exert the drug effect, and are widely used as the carriers for transdermal absorption of macromolecular drugs and water-soluble and fat-soluble drugs.
The invention aims to solve the problems, and provides a triamcinolone acetonide acetate composition spray which consists of triamcinolone acetonide acetate, a somatostatin modifier, DSPE-PEG2000, phospholipid, cholesterol, a liposome membrane regulator and a pharmaceutically acceptable carrier;
wherein the spray comprises a carrier, the carrier comprises DSPE-PEG2000, phospholipid, cholesterol and liposome membrane regulator, the particle size range is 40-100 nm, and at least 80% of the carrier particle size is uniform.
In particular, the pharmaceutically acceptable carrier is conventional. In some examples of the invention, the pharmaceutically acceptable carrier includes a solvent, a phosphate buffer, and the like.
Specifically, in some examples of the invention, the solvent is a conventional short chain alcohol, including ethanol, propylene glycol.
Preferably, the somatostatin modifier in the spray is DSPE-PEG2000-SST, and the synthesis process is as follows: stirring SST and DSPE-PEG2000-BTC (molar ratio is 1: 2) in 50mL DMSO at 4 ℃, adjusting the pH to 7.0-7.5 by triethylamine, dialyzing for 24h after the reaction is finished, and freeze-drying to obtain the DSPE-PEG 2000-SST. The PEG modification of the SST can improve the in vivo bioactivity of the SST, prolong the administration time, improve the stability of a carrier and enhance the encapsulation efficiency.
Preferably, the dosage of triamcinolone acetonide acetate in the spray is 5-500mg/mL, the dosage of somatostatin modifier is 0.2-20 mg/mL, the dosage of DSPE-PEG2000 is 0.04-0.6 g/mL, the dosage of phospholipid is 0.03-0.5 g/mL, the dosage of cholesterol is 0.005-0.2 g/mL, and the dosage of liposome membrane regulator is 0.004-0.15 g/mL.
Preferably, the phospholipid in the spray is selected from soybean lecithin and egg yolk lecithin.
Preferably, the liposome membrane regulator in the spray is selected from one or more of sodium cholate, sodium deoxycholate and Triton-100.
More preferably, the liposome membrane regulator in the spray is sodium cholate.
Preferably, the carrier particle size in the spray is in the range of 40-100 nm, and at least 90% of the particles are uniform. The carrier has small and uniform particle size, and is helpful for reducing the metabolism of the drug in vivo and improving the bioavailability.
The invention also provides a preparation method of the triamcinolone acetonide composition spray, which comprises the following steps:
(1) SST and DSPE-PEG2000 are prepared into DSPE-PEG2000-SST by adopting a chemical coupling method;
(2) dissolving the DSPE-PEG2000-SST obtained in the step (1), the DSPE-PEG2000, phospholipid, cholesterol and liposome membrane regulator in an organic solvent, and rotationally evaporating the organic solvent to form a uniform film A;
(3) dissolving triamcinolone acetonide acetate in water to obtain a solution B;
(4) adding the solution B into the film A, carrying out ultrasonic atomization at the frequency of 1.2-2.3 MHz and the temperature of 15-50 ℃ by an ultrasonic atomizer, introducing into a phosphate buffer solution which is stirring, filtering, and sterilizing to obtain the spray;
wherein, in the step (2), the dosage of the DSPE-PEG2000-SST is 0.2-20 mg/mL, the dosage of the DSPE-PEG2000 is 0.04-0.6 g/mL, the dosage of the phospholipid is 0.03-0.5 g/mL, the dosage of the cholesterol is 0.005-0.2 g/mL, and the dosage of the liposome membrane regulator is 0.004-0.15 g/mL;
the dosage of the triamcinolone acetonide acetate in the step (3) is 5-500 mg/mL.
Preferably, the frequency of ultrasonic atomization in the step (4) in the preparation method is 1.5-1.7MHz, and the temperature of ultrasonic atomization is 20-40 ℃. Ultrasonic atomization is helpful for the uniform particle size of the carrier, and when the ultrasonic intensity is too high, the prepared carrier is partially destroyed, and the encapsulation efficiency is reduced. When the ultrasonic intensity is too small, the carrier cannot be sufficiently dispersed in the buffer solution, resulting in an increase in particle size. The encapsulation efficiency is reduced due to overhigh ultrasonic atomization temperature; the temperature of ultrasonic atomization is too low, which is not favorable for the dispersion of the carrier in the buffer solution.
The mechanism of action of SST in the treatment of arthritis: kolasinski considers that somatostatin dilates blood vessels by acting directly on vascular smooth muscle and causing mast cells to release histamine and enhance local vascular permeability indirectly, and treats arthritic diseases such as rheumatoid Arthritis (Kolasinski SL, Haines KA, Siegel EL, et al. neuropeps and inflammation: a monomeric analogon a selective anti-inflammatory of neutral activity by sub-stance P. Arthritis Rheum, 1992,35: 369-372). Matucci suggests that the effect of somatostatin in treating arthritis may also be related to inhibition of substance P production and inhibition of SP release from involved nerve endings (delta fiber and C fiber), and has the functions of inhibiting prostaglandin release from joint synovial cells and inhibiting activation and chemotaxis of macrophages (Matucci-Cerinic M. sensory neuronopiledes and rhematic disease. Rheum Dis Clin N Am, 1993,19: 975-976). Somatostatin acts on different kinds of cells and tissues in the joint, the main site of action is the sensory nerve endings, while Fioravant speculates that another site of action of Somatostatin is the synovial cells, inhibiting the proliferation of synovial cells, as well as inhibiting the proliferation of lymphocytes and the chemotaxis of monocytes (mainly lymphocytes) (Fioravanti A, Govoni M, La Montagna G, et al. somatotatin 14 and joint inflammation: evidence for intracellular differentiation of committed administration in renal tissue, Drugs Expt Clin Res, 1995,21: 97-103). The mechanism of treating arthritis by triamcinolone acetonide is as follows: triamcinolone acetonide can effectively inhibit the degradation process of osteoarthritis, and can inhibit the activity of metalloprotease to improve the symptoms of osteoarthritis. The combined administration of SST and triamcinolone acetonide can treat arthritis from different mechanisms, and has synergistic effect.
Marco et al reported that somatostatin treatment was statistically significant and dose-dependent in knee swelling, a shorter effect than triamcinolone acetonide. However, triamcinolone acetonide in successive doses decreased in both magnitude and duration of anti-inflammatory activity, while the effect of Somatostatin remained unchanged with each successive treatment (Marco Matucci-Cerinic MD, France sco Borrelli MS, et al. In addition, SST has mild effect, needs to be injected for a plurality of times at intervals and then slowly accumulated to take effect, and has high price. The SST and triamcinolone acetonide combined drug can improve the defect of slow accumulation and effect of SST and solve the problem of the reduction of the duration time of the anti-inflammatory activity of triamcinolone acetonide. The PEG modification of the SST can improve the in vivo biological activity of the SST, prolong the administration time and solve the problem of rapid in vivo metabolism of the polypeptide.
The invention creatively combines triamcinolone acetonide and somatostatin for administration, provides a triamcinolone acetonide composition spray, can relieve arthralgia and swelling from different mechanisms, has synergistic effect, and has quick response and lasting treatment effect. In addition, the invention also provides a preparation method of the triamcinolone acetonide composition spray, and the prepared transfersome has small and uniform particle size and good transdermal effect.
Detailed Description
The following examples are further illustrative of the present invention and are in no way intended to limit the scope of the invention. The present invention is further illustrated in detail below with reference to examples, but it should be understood by those skilled in the art that the present invention is not limited to these examples and the preparation method used. Also, equivalent substitutions, combinations, improvements or modifications of the invention may be made by those skilled in the art based on the description of the invention, but these are included in the scope of the invention.
Example 1
Composition (A) Mass/volume
Triamcinolone acetonide acetate 50mg
DSPE-PEG2000-SST 200 mg
DSPE-PEG2000 0.6 kg
Egg yolk lecithin 0.5 kg
Cholesterol 0.2 kg
Deoxycholic acid sodium salt 0.15 kg
10 mmol of phosphate buffer (pH6.5) to 1L
(1) Stirring SST and DSPE-PEG2000-BTC (the molar ratio is 1: 2) in 50 mLDMSO at the temperature of 4 ℃, adjusting the pH to 7.0-7.5 by triethylamine, dialyzing for 24h after the reaction is finished, and freeze-drying to obtain DSPE-PEG 2000-SST;
(2) weighing the DSPE-PEG2000-SST prepared in the step (1), dissolving the DSPE-PEG2000, yolk lecithin, cholesterol and deoxysodium cholate in 500 mL of propylene glycol, and rotationally evaporating the propylene glycol to form a uniform film A;
(3) dissolving triamcinolone acetonide acetate with 50mL of ultrapure water to obtain a solution B;
(4) adding the solution B into the film A, carrying out ultrasonic atomization at the frequency of 1.7MHz and the temperature of 25 ℃ by an ultrasonic atomizer, introducing into a phosphate buffer solution which is stirring, stirring for 1h, carrying out rough filtration on a liquid medicine by a bag filter, and carrying out sterilization by a 0.20-micron microporous filter membrane to obtain the spray.
The average particle size of the encapsulated drug carrier transfersomes in the spray is measured to be 50 nm, the spray is spherical, the uniformity is 95 percent, the encapsulation rate is 92.5 percent, the spray is stored for 180 days at the low temperature of 2-8 ℃, and the solution is uniform, stable and free from layering and deterioration.
Example 2
Composition (I) Mass/volume
Triamcinolone acetonide acetate 2000mg
DSPE-PEG2000-SST 100 mg
DSPE-PEG2000 0.04 kg
Soybean lecithin 0.25 kg
Cholesterol 0.1 kg
Cholesterol acid sodium salt 0.02 kg
10 mmol of phosphate buffer solution (pH7.5) was added 1L
(1) Stirring SST and DSPE-PEG2000-BTC (the molar ratio is 1: 2) in 50mL DMSO at 4 ℃, adjusting the pH to 7.0-7.5 by triethylamine, dialyzing for 24h after the reaction is finished, and freeze-drying to obtain DSPE-PEG 2000-SST;
(2) weighing the DSPE-PEG2000-SST prepared in the step (1), dissolving the DSPE-PEG2000, the soybean lecithin, the cholesterol and the sodium cholate in 500 mL of ethanol, and performing rotary evaporation on the ethanol to form a uniform film A;
(3) dissolving triamcinolone acetonide acetate with the weight of 50mL of ultrapure water to obtain a solution B;
(4) adding the solution B into the film A, carrying out ultrasonic atomization at the frequency of 2.3 MHz and the temperature of 15 ℃ by an ultrasonic atomizer, introducing into a phosphate buffer solution which is stirring, stirring for 1h, carrying out rough filtration on a liquid medicine by a bag filter, and carrying out sterilization by a 0.20-micron microporous filter membrane to obtain the spray.
The average grain diameter of the encapsulated drug carrier transfersomes in the spray is measured to be 40 nm, the spray is spherical, the uniformity is 92 percent, the encapsulation rate is 94.5 percent, the spray is stored for 180 days at the low temperature of 2-8 ℃, and the solution is uniform, stable and free from layering and deterioration.
Example 3
Composition (I) Mass/volume
Triamcinolone acetonide acetate 5000mg
DSPE-PEG2000-SST 2 mg
DSPE-PEG2000 0.3 kg
Soybean lecithin 0.03 kg
Cholesterol 0.005 kg
Triton-100 0.004 kg
10 mmol of phosphate buffer (pH7.0) to 1L
(1) Stirring SST and DSPE-PEG2000-BTC (the molar ratio is 1: 2) in 50mL DMSO at 4 ℃, adjusting the pH to 7.0-7.5 by triethylamine, dialyzing for 24h after the reaction is finished, and freeze-drying to obtain DSPE-PEG 2000-SST;
(2) weighing the DSPE-PEG2000-SST prepared in the step (1) according to the weight, dissolving the DSPE-PEG2000, the soybean lecithin, the cholesterol and the Triton-100 in 500 mL of ethanol according to the weight, and performing rotary evaporation on the ethanol to form a uniform film A;
(3) dissolving triamcinolone acetonide acetate with the weight of 50mL of ultrapure water to obtain a solution B;
(4) adding the solution B into the film A, carrying out ultrasonic atomization at the frequency of 1.2 MHz and the temperature of 50 ℃ by an ultrasonic atomizer, introducing into a phosphate buffer solution which is stirring, stirring for 1h, carrying out rough filtration on a liquid medicine by a bag filter, and carrying out sterilization by a 0.20-micron microporous filter membrane to obtain the spray.
The average particle size of the encapsulated drug carrier transfersomes in the spray is determined to be 100 nm, the spray is spherical, the uniformity is 92%, the encapsulation rate is 93.5%, the spray is stored for 180 days at the low temperature of 2-8 ℃, and the solution is uniform, stable and free from layering and deterioration.
Example 4
Composition (I) Mass/volume
Triamcinolone acetonide acetate 500mg
DSPE-PEG2000-SST 150 mg
DSPE-PEG2000 0.4 kg
Egg yolk lecithin 0.1 kg
Cholesterol 0.12 kg
Cholesterol acid sodium salt 0.09 kg
10 mmol of phosphate buffer (pH7.0) to 1L
(1) Stirring SST and DSPE-PEG2000-BTC (the molar ratio is 1: 2) in 50mL DMSO at 4 ℃, adjusting the pH to 7.0-7.5 by triethylamine, dialyzing for 24h after the reaction is finished, and freeze-drying to obtain DSPE-PEG 2000-SST;
(2) weighing the DSPE-PEG2000-SST prepared in the step (1), dissolving the DSPE-PEG2000, yolk lecithin, cholesterol and sodium cholate in 500 mL of propylene glycol, and rotationally evaporating the propylene glycol to form a uniform film A;
(3) dissolving triamcinolone acetonide acetate with the weight of 50mL of ultrapure water to obtain a solution B;
(4) adding the solution B into the film A, carrying out ultrasonic atomization at the frequency of 1.6 MHz and the temperature of 20 ℃ by an ultrasonic atomizer, introducing into a phosphate buffer solution which is stirring, stirring for 1h, carrying out rough filtration on a liquid medicine by a bag filter, and carrying out sterilization by a 0.20-micron microporous filter membrane to obtain the spray.
The average particle size of the encapsulated drug carrier transfersomes in the spray is measured to be 70 nm, the spray is spherical, the uniformity is 99 percent, the encapsulation rate is 94.5 percent, the spray is stored for 180 days at the low temperature of 2-8 ℃, and the solution is uniform, stable and free from layering and deterioration.
Example 5
Composition (I) Mass/volume
Triamcinolone acetonide acetate 3250mg
DSPE-PEG2000-SST 50 mg
DSPE-PEG2000 0.1 kg
Egg yolk lecithin 0.15 kg
Cholesterol 0.05 kg
Deoxycholic acid sodium salt, sodium cholate 0.08 kg
10 mmol of phosphate buffer (pH7.0) to 1L
(1) Stirring SST and DSPE-PEG2000-BTC (the molar ratio is 1: 2) in 50mL DMSO at 4 ℃, adjusting the pH to 7.0-7.5 by triethylamine, dialyzing for 24h after the reaction is finished, and freeze-drying to obtain DSPE-PEG 2000-SST;
(2) weighing the DSPE-PEG2000-SST prepared in the step (1), dissolving the DSPE-PEG2000, yolk lecithin, cholesterol, deoxysodium cholate and sodium cholate in 500 mL of ethanol, and performing rotary evaporation on the ethanol to form a uniform film A;
(3) dissolving triamcinolone acetonide acetate with the weight of 50mL of ultrapure water to obtain a solution B;
(4) adding the solution B into the film A, carrying out ultrasonic atomization at the frequency of 1.5 MHz and the temperature of 40 ℃ by an ultrasonic atomizer, introducing into a phosphate buffer solution which is stirring, stirring for 1h, carrying out rough filtration on a liquid medicine by a bag filter, and carrying out sterilization by a 0.20-micron microporous filter membrane to obtain the spray.
The average grain diameter of the carrier transfersomes of the encapsulated drug in the spray is determined to be 80 nm, the spray is spherical, the uniformity is 99 percent, the encapsulation rate is 94.5 percent, the spray is stored for 180 days at the low temperature of 2-8 ℃, and the solution is uniform, stable and free from layering and deterioration.
Example 6 in vitro permeability test
Adopting an in vitro skin method: the rat skin was naturally fixed on a modified Franz diffusion cell (diffusion area S ═ 2.5 cm) 2 ) The skin surface was facing the dosing chamber and the inner layer of the skin was in intimate contact with a receiving fluid (i.e. saline, 6 mL receiving reservoir volume). And uniformly spraying the sample on the surface of the rat skin. Placing the diffusion cell in a constant temperature magnetic stirrer, controlling the temperature at 37 + -1 deg.C and the rotation speed at 30 r.min -1 . Stirring for 12h, taking 10mL of receiving solution, filtering the receiving solution through a 0.45-micrometer microporous filter membrane, injecting by HPLC, and determining the content of somatostatin and triamcinolone acetonide in the receiving solution. The transdermal rate is calculated according to the formula, and the transdermal absorption rate is equal to transdermal quantity/administration quantity multiplied by 100%. The in vitro skin penetration rates of examples 1-5 were determined to be 45.2%, 47.2%, 43.4%, 50.8%, 51.2%, respectively.
EXAMPLE 7 Effect of treating rheumatoid arthritis
(1) Animal model and group preparation
The cleaning grade female and male Wistar rats are 110, 4-6 weeks old and have the weight (180 +/-10) g. Adopting a type II collagen induction rat arthritis model, dissolving chicken type II collagen in 0.01mol/L acetic acid solution with the concentration of 5g/L, mixing with equal volume of Freund's complete adjuvant to prepare an emulsion, adding BCG vaccine, and carrying out intradermal injection immunization on the tail and the back of 100 experimental rats; 14d hind paw subcutaneous booster immunization 1 time. Randomly grouping after the model is completed: model group, no drug administration; somatostatin group administered: injecting somatostatin injection in joint 1 time per week; triamcinolone acetonide administration group: the triamcinolone acetonide injection is injected in the joints for 1 time per week; spray administration group: the triamcinolone acetonide composition spray is sprayed to joints twice a day, and the dosage is 1 mL each time. After 5 weeks of treatment, anesthetizing the rat, drawing blood, separating serum, preserving biochemical detection at-70 ℃ for later use, inactivating a part of the blood at 56 ℃ for 30min, and filtering and sterilizing to obtain serum containing medicine for later use; meanwhile, the joint cavity is opened, the synovial tissue is taken out and fixed by 70% alcohol, and a tissue section is reserved.
(2) Detection item and detection method
a. The joint swelling index was evaluated by a 0-4 grade joint score. 0 minute: no arthritis; 1 minute: mild swelling of the toe joints; and 2, dividing: swelling of the little toe joint and the plantar aspect; and 3, dividing: paw swelling below the ankle joint; and 4, dividing: swelling of all joints including the ankle; observation point 1 was before dosing, and 1 time per week and 5 times thereafter.
b. The biochemical assays used IL-1 β (TPI Inc IR 043) and TNF- α (TPI Inc IR 0820) Elisa kits and a Wellscan MK type II enzyme target.
c. Measurement of synovial cell proliferation with serum containing drug A model group of rat primary synovial cell suspension (2X 10) was prepared using culture solution containing 10% calf serum-DMEM 6 /mL), added to a 96-well plate at 100. mu.L per well, and incubated at 37 ℃ with 5% CO 2 Culturing for 24h in an incubator, and culturing for 24h by changing a DMEM culture solution without serum after adherence, so that the cells are relatively synchronized. Serum-free culture solution was aspirated, 100 μ L each of 10% serum-containing DMEM was added, culture was continued for 68 hours, 20 μ L of MTT (5 mg/L) solution was added to each well, culture was continued for 4 hours, the supernatant was discarded, 200 μ L of DMSO was added, OD (λ ═ 490 nm) (optical density, i.e., OD value) per well was measured with a microplate reader after shaking for 3 minutes, and the results were expressed as the mean value of 6 wells per group using normal rat serum as a control group.
(3) Results
a. Index of joint swelling
Table 1 shows the results of the joint swelling index measurement, and it can be seen from the table that the somatostatin-administered group: when the dosage is lower, the effect of relieving the joint swelling is weak and slow, the effect of relieving the joint swelling is enhanced along with the increase of the dosage, and the action time of continuous treatment is prolonged. Triamcinolone acetonide administration group: when the dosage is lower, the effect of relieving the joint swelling is weak, the effect of relieving the joint swelling is enhanced along with the increase of the dosage, but the effect of continuous treatment effect is reduced, such as: the dose of 45.5 mg or 70 mg was 1 week after administration to relieve joint swelling, but the sustained therapeutic effect was reduced after 4 weeks after administration. Spray administration group: cumulative weekly dose of the spray prepared in example 1: 2.8 mg of somatostatin and 0.7 mg of triamcinolone acetonide; cumulative weekly dose of the spray prepared in example 3: somatostatin 0.028 mg, triamcinolone acetonide 70 mg; cumulative weekly dose of the spray prepared in example 5: somatostatin 0.7 mg, triamcinolone acetonide 45.5 mg. The spray prepared in the examples 1, 3 and 5 has quick response and lasting treatment effect on relieving the joint swelling. The results show that the triamcinolone acetonide and somatostatin combined drug has a synergistic effect on relieving joint pain and swelling, and has the advantages of quick response and lasting continuous treatment effect.
TABLE 1 determination of joint swelling index (x. + -.s)
Figure DEST_PATH_IMAGE002
b. Serum TNF-alpha, IL-1 beta
The secretion of TNF-alpha and IL-1 beta by inflammation is a centrally located pro-inflammatory cytokine in the pathogenesis of rheumatoid inflammation. Table 2 shows the results of the measurement of serum TNF- α and IL-1 β, and it can be seen from the table that the TNF- α levels of the somatostatin administration group, the triamcinolone acetonide administration group and the spray administration group were all decreased to different degrees compared to the model group, and the decrease degree of the spray administration group was the highest; in addition, IL-1. beta. levels were also reduced in the somatostatin-administered group and the spray-administered group. The results show that the combined use of the triamcinolone acetonide and the somatostatin has synergistic effect on relieving the joint pain and the swelling, has lasting treatment effect, and is superior to the single injection of the triamcinolone acetonide or the somatostatin.
TABLE 2 serum TNF-. alpha.IL-1. beta. assay (x. + -. s)
Figure DEST_PATH_IMAGE004
c. Histology of synovial membrane of rat knee joint
Normal rat synovium and articular surface have no pathological changes, the synovium lining layer is composed of 1-2 layers of synovium cells and is partially discontinuous, the lower layer of the synovium lining layer is adipose cell nucleus blood vessels, and inflammatory cell infiltration, fibroblasts and collagen fibers are not seen. The synovial tissue in the knee joint of the model group animal swells, the upper layer of the synovial membrane thickens, synovial cells increase and increase in number, the synovial membrane has obvious inflammation, blood vessel congestion in the connective tissue in the synovial tissue, blood capillaries obviously increase, fibrous tissue is loose, more chronic inflammatory cells are infiltrated, and the articular cartilage can be seen to have flaky necrotic foci with different sizes and irregular edges. Compared with the model group, the pathological changes of the somatostatin administration group, the triamcinolone acetonide administration group and the spraying agent administration group are reduced to different degrees, mainly expressed in that the number of inflammatory cells in joint soft tissues is reduced; angiogenesis, articular cartilage destruction, and edge deformity are rare. Table 3 shows the effect of the drug-containing serum on synovial cell proliferation, and the OD values of the spray administration group and the blank group are the closest, which indicates that the combined use of triamcinolone acetonide and somatostatin has synergistic effect in relieving joint pain and swelling, and the sustained therapeutic effect is lasting, which is superior to the single injection of triamcinolone acetonide or somatostatin.
TABLE 3 Effect of drug-containing sera on synovial cell proliferation (x. + -. s)
Figure DEST_PATH_IMAGE006

Claims (9)

1. A triamcinolone acetonide acetate composition spray is characterized by consisting of triamcinolone acetonide acetate, somatostatin modifiers, DSPE-PEG2000, phospholipid, cholesterol, a liposome membrane regulator and a pharmaceutically acceptable carrier;
wherein the spray comprises a carrier, the carrier comprises DSPE-PEG2000, phospholipid, cholesterol and liposome membrane regulator, the particle size range is 40-100 nm, and at least 80% of the carrier particle size is uniform.
2. The spray according to claim 1, wherein the somatostatin modification in the spray is DSPE-PEG 2000-SST.
3. The spray according to claim 1, wherein the amount of triamcinolone acetonide acetate in the spray is 5-500mg/mL, the amount of somatostatin modification is 0.2-20 mg/mL, the amount of DSPE-PEG2000 is 0.04-0.6 g/mL, the amount of phospholipid is 0.03-0.5 g/mL, the amount of cholesterol is 0.005-0.2 g/mL, and the amount of liposome membrane regulator is 0.004-0.15 g/mL.
4. The spray according to claim 1, wherein the phospholipid in the spray is selected from the group consisting of soy lecithin and egg yolk lecithin.
5. The spray according to claim 1, wherein the liposome membrane regulator in the spray is selected from one or more of sodium cholate, sodium deoxycholate, and Triton-100.
6. The spray according to claim 5, wherein the liposome membrane regulator in the spray is sodium cholate.
7. The spray according to claim 1, wherein the carrier in the spray has a particle size in the range of 50-80 nm, and at least 90% of the particles are uniform.
8. A process for preparing the spray of claim 1, comprising the steps of:
(1) SST and DSPE-PEG2000 are prepared into DSPE-PEG2000-SST by adopting a chemical coupling method;
(2) dissolving the DSPE-PEG2000-SST obtained in the step (1), the DSPE-PEG2000, phospholipid, cholesterol and liposome membrane regulator in an organic solvent, and rotationally evaporating the organic solvent to form a uniform film A;
(3) dissolving triamcinolone acetonide acetate in water to obtain a solution B;
(4) adding the solution B into the film A, performing ultrasonic atomization at the frequency of 1.2-2.3 MHz and the temperature of 15-50 ℃ by an ultrasonic atomizer, introducing into a phosphate buffer solution under stirring, filtering, and sterilizing to obtain the spray;
wherein, in the step (2), the dosage of the DSPE-PEG2000-SST is 0.2-20 mg/mL, the dosage of the DSPE-PEG2000 is 0.04-0.6 g/mL, the dosage of the phospholipid is 0.03-0.5 g/mL, the dosage of the cholesterol is 0.005-0.2 g/mL, and the dosage of the liposome membrane regulator is 0.004-0.15 g/mL;
the dosage of the triamcinolone acetonide acetate in the step (3) is 5-500 mg/mL.
9. The method of claim 8, wherein the ultrasonic atomization is carried out at a frequency of 1.5 to 1.7MHz and at a temperature of 20 to 40 ℃.
CN202110131270.6A 2021-01-30 2021-01-30 Triamcinolone acetonide acetate composition spray and preparation method thereof Active CN114831939B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN202110131270.6A CN114831939B (en) 2021-01-30 2021-01-30 Triamcinolone acetonide acetate composition spray and preparation method thereof

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN202110131270.6A CN114831939B (en) 2021-01-30 2021-01-30 Triamcinolone acetonide acetate composition spray and preparation method thereof

Publications (2)

Publication Number Publication Date
CN114831939A true CN114831939A (en) 2022-08-02
CN114831939B CN114831939B (en) 2024-06-07

Family

ID=82561384

Family Applications (1)

Application Number Title Priority Date Filing Date
CN202110131270.6A Active CN114831939B (en) 2021-01-30 2021-01-30 Triamcinolone acetonide acetate composition spray and preparation method thereof

Country Status (1)

Country Link
CN (1) CN114831939B (en)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN117357480A (en) * 2023-12-08 2024-01-09 北京中科利华医药研究院有限公司 Composition for needleless percutaneous injection

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1327382A (en) * 1998-12-23 2001-12-19 伊迪亚股份公司 Improved formulation for topical non-invasive application in vivo
CN1543932A (en) * 2003-11-25 2004-11-10 华中科技大学 Application of solid liposome nanoparticle for preparing pharmaceutical used for ion leading-in trandermal therapy
CN101439182A (en) * 2008-12-18 2009-05-27 北京大学 Somatostatin receptor mediated tumor-targeted medicament composition
CN104173285A (en) * 2013-05-23 2014-12-03 上海市第七人民医院 Preparation method for triamcinolone acetonide acetate ultradeformable nanometer lipid vesicle spraying agent and application to target hypertrophic scars
CN109789215A (en) * 2016-09-27 2019-05-21 卡姆拉斯公司 Preparation containing Somat receptor stimulating agent

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1327382A (en) * 1998-12-23 2001-12-19 伊迪亚股份公司 Improved formulation for topical non-invasive application in vivo
CN1543932A (en) * 2003-11-25 2004-11-10 华中科技大学 Application of solid liposome nanoparticle for preparing pharmaceutical used for ion leading-in trandermal therapy
CN101439182A (en) * 2008-12-18 2009-05-27 北京大学 Somatostatin receptor mediated tumor-targeted medicament composition
CN104173285A (en) * 2013-05-23 2014-12-03 上海市第七人民医院 Preparation method for triamcinolone acetonide acetate ultradeformable nanometer lipid vesicle spraying agent and application to target hypertrophic scars
CN109789215A (en) * 2016-09-27 2019-05-21 卡姆拉斯公司 Preparation containing Somat receptor stimulating agent

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
GERBEN A. KONING等: "Targeting of Angiogenic Endothelial Cells at Sites of Inflammation by Dexamethasone Phosphate–Containing RGD Peptide Liposomes Inhibits Experimental Arthritis", 《ARTHRITIS & RHEUMATISM》 *
GREGOR CEVC等: "Biological activity and characteristics of triamcinolone-acetonide formulated with the self-regulating drug carriers, Transfersomes", 《BIOCHIMICA ET BIOPHYSICA ACTA》 *
MARC0 MATUCCI-CERINIC等: "SOMATOSTATIN-INDUCED MODULATION OF INFLAMMATION IN EXPERIMENTAL ARTHRITIS", 《ARTHRITIS & RHEUMATISM》 *

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN117357480A (en) * 2023-12-08 2024-01-09 北京中科利华医药研究院有限公司 Composition for needleless percutaneous injection
CN117357480B (en) * 2023-12-08 2024-03-29 北京中科利华医药研究院有限公司 Composition for needleless percutaneous injection

Also Published As

Publication number Publication date
CN114831939B (en) 2024-06-07

Similar Documents

Publication Publication Date Title
CN1273187C (en) Insulin preparations, which donot contain any zinc or only small quantity of zinc of improved stability
JP3919212B2 (en) Wound healing treatment for fibrosis
JP2007515439A (en) Medical lipolysis of fat accumulation
CN1997383B (en) Aequorin-containing compositions and methods of using same
CN114831939B (en) Triamcinolone acetonide acetate composition spray and preparation method thereof
KR102490400B1 (en) Composition for treating or prventing osteoarthritis
CA2100720C (en) Medicinal composition comprising tcf-ii
JPH01151514A (en) Treating and preventive agent for neuropathy
US8703704B2 (en) Method for growing, restoring and regenerating hair
US20080318863A1 (en) Ischemia therapeutic agent
US20100297100A1 (en) Composition for Treating Retinopathy or Glaucoma Comprising Thrombin Derived Peptides
CN102274182B (en) Multivesicular liposome containing exenatide, preparation method and application thereof
CN116211827B (en) Teriparatide solid lipid nanoparticle and preparation method and application thereof
WO2015073958A1 (en) Ephedra alata extracts and methods of use thereof
TWI395592B (en) Method for controlled release of parathyroid hormone from encapsulated poly (lactic-co-glycolic acid) microspheres
JP3652865B2 (en) Mixture of lactic acid condensate and composition containing the same
CN111671718B (en) Flurbiprofen axetil injection and preparation method thereof
CN115252621B (en) Application of small molecular compound in preparation of medicine for treating osteoarthritis
CN110464713B (en) Preparation method of skin penetration patch for resisting rheumatoid arthritis by using Qisonol
Jing et al. Preparation of Baicalein Long-Circulating Liposome and Its Effect on Bone Implantation in Diabetic Rats
KR20240086947A (en) Peptide for cartilage regeneration and uses thereof
CN117899006A (en) Composite hydrogel loaded with disulfiram nano vesicles and application of composite hydrogel in preparation of diabetic skin wound healing drugs
RU2277413C2 (en) Method for treating secondary dystrophia corneae
EP0672420A1 (en) Wound remedy
KR20240086946A (en) Peptide for cartilage regeneration and uses thereof

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
GR01 Patent grant