WO2014027360A1 - Concentré de facteurs de croissance pour traiter des rides faciales - Google Patents

Concentré de facteurs de croissance pour traiter des rides faciales Download PDF

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Publication number
WO2014027360A1
WO2014027360A1 PCT/IN2013/000264 IN2013000264W WO2014027360A1 WO 2014027360 A1 WO2014027360 A1 WO 2014027360A1 IN 2013000264 W IN2013000264 W IN 2013000264W WO 2014027360 A1 WO2014027360 A1 WO 2014027360A1
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Prior art keywords
growth factor
concentrate
approximately
facial wrinkles
gfc
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PCT/IN2013/000264
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English (en)
Inventor
Satish Mahadeorao Totey
Kaushal Piyush SHAH
Lyle Carl FONSECA
Rachana Rajiv MANIYAR
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Kasiak Research Pvt. Ltd.
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Publication of WO2014027360A1 publication Critical patent/WO2014027360A1/fr

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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
    • A61K38/1858—Platelet-derived growth factor [PDGF]
    • A61K38/1866—Vascular endothelial growth factor [VEGF]
    • 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
    • A61K38/1808—Epidermal growth factor [EGF] urogastrone
    • 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
    • A61K38/1825—Fibroblast growth factor [FGF]
    • 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
    • A61K38/1841—Transforming growth factor [TGF]
    • 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
    • A61K38/1858—Platelet-derived growth factor [PDGF]

Definitions

  • a growth factor concentrate for treating facial wrinkles A growth factor concentrate for treating facial wrinkles
  • This invention relates to a growth factor concentrate for treating facial wrinkles.
  • nasolabial wrinkles commonly known as "smile lines” or “laugh lines,” are facial features. They are the two skin folds that run from each side of the nose to the corners of the mouth. They separate the cheeks from the upper lip. The term derives from Latin nasus for "nose” and labium for "lip".
  • nasolabial wrinkles develop as we age and can be one of the first visible signs of facial aging. That's why nasolabial wrinkles are also one of the first areas of the face targeted by anti-aging or facial rejuvenation procedures. With ageing the fold may become more pronounced and may become cosmetically undesirable to some. The most significant factors that contribute to the prominence of the fold are excess skin, skin thinning, excess cheek fat, and ptosis of cheek fat. Methods of addressing facial wrinkles range from excision and re-draping of excess skin to methods of skin resurfacing including dermabrasion, laser ablation, and chemical peels. Each of these modalities has limitations. Implantation of a filler substance into a deficient area is another cosmetic solution. Many materials have been tried for the correction of soft tissue defects and deficits. However, the use of most of these substances has met with difficulties, including impermanence, foreign body reaction, unnatural texture, and possible disease transmission. In addition, most of the products currently available remain relatively expensive.
  • Collagen currently is used widely for treating facial wrinkles.
  • Available collagen consists of purified bovine collagen comprising 95% type I collagen and 5% type III collagen.
  • glutaraldehyde for cross-linking such collagen slows resorption; however, the substance nonetheless eventually is degraded by the host.
  • collagen works well for the correction of fine lines, results for deeper furrows are less impressive.
  • the major drawback to the use of collagen is the short duration of the response. Within 3 to 6 months the collagen is resorbed completely, and the improvement is lost. Also, because the collagen is bovine-derived, pre- testing for an allergic response is required.
  • FDA Food and Drug Administration
  • the result is generally predictable because the substance is nonreactive, but the resulting feel of the tissue is somewhat unnatural.
  • the substance is rigid and is not available in an injectable form.
  • infection remains a potential problem because the substance is a foreign body. Because this substance does not become integrated into the host tissue, it is generally easy to remove.
  • PLLA injectable poly-L-lactic acid
  • an intra-dermally, sub-dermally or topically administrate growth factor concentrate derived from approximately 250 ⁇ ⁇ ⁇ 6 to 5000 x l O 6 human platelets per ml for treating facial wrinkles, the concentrate comprising approximately 900 to 2000 pg/ml of Epidermal growth factor (EGF), 30 to 300 pg/ml of Vascular Endothelial growth factor(VEGF), 20 to 100 pg/ml of Basic fibroblast growth factor (b-FGF), 40000 to 120000 pg/ml of Transforming growth factor- ⁇ (TGF- ⁇ ) and 200000 to 600000 pg/ml of Platelet Derived growth factor-AB (PDGF-AB).
  • EGF Epidermal growth factor
  • VEGF Vascular Endothelial growth factor
  • b-FGF Basic fibroblast growth factor
  • TGF- ⁇ Transforming growth factor- ⁇
  • PDGF-AB Platelet Derived growth factor-AB
  • a lyophilized, intra- dermally, sub-dermally or topically administrable growth factor concentrate derived from approximately 250 ⁇ 10 6 to 5000 x lO 6 human platelets per ml for treating facial wrinkles, the concentrate comprising approximately 900 to 2000 pg/ml of EGF, 30 to 300 pg/ml of VEGF, 20 to 100 pg/ml of b-FGF, 40000 to 120000 pg/ml of TGF- ⁇ and 200000 to 600000 pg/ml of PDGF-AB suspended in an isotonic medium along with excipients for lyophilisation.
  • a therapeutic composition for intra-dermal, sub-dermal or topical administration for treatment of facial wrinkles comprising the growth factor concentrate in combination with supplemental constituents including blood, saline, silver nanoparticles, hyaluronic acid, immuno-modulatory peptides, growth factors, hormones, antibiotics, monoclonal antibodies, recombinant receptors, carriers or combinations thereof.
  • a method of treatment of facial wrinkles comprising injecting a growth factor concentrate into the facial wrinkles, wherein the growth factor concentrate is derived from approximately 625 x lO 6 human platelets per ml, the concentrate comprising approximately 500 to 1000 pg/ml of EGF, 10 to 20 pg/ml of VEGF, 10 to 25 pg/ml of basic b-FGF, 20000 to 30000 pg/ml of TGF- ⁇ and 60000 to 150000 pg/ml of PDGF-AB.
  • FIG. 1A and FIG. I B respectively are photographic representations of a study subjects' nasolabial wrinkles before and at three months after treatment with the GFC as per Example 2.
  • FIG. 2 shows the Wrinkle Severity Rating Scale (WSRS) scores of study subjects, before treatment and at one month, two months and three months after treatment with the GFC as per Example 2.
  • the wrinkles are scored on a scale of 0-4 with Grade 0 being no nasolabial wrinkles and Grade 4 being deepest nasolabial wrinkles.
  • FIG. 3 shows the Mean WSRS scores of twenty study subjects, before treatment and at one month, two months and three months after treatment with the GFC as per Example 2.
  • the wrinkles are scored on a scale of 0-4 with Grade 0 being no nasolabial wrinkles and Grade 4 being deepest nasolabial wrinkles.
  • growth factor concentrate or "GFC” as used in the specification refers to a standardized concentration of growth factors prepared according to an embodiment of the invention, the growth factors being derived from the cryo-stimulation of platelets which have been counted, the platelets being sourced from human blood.
  • the GFC also contains cytokines, chemokines, adhesive proteins and other modulatory peptides.
  • multiple electrolyte isotonic solution as used in the specification in the context of the invention comprises sodium chloride, (NaCl); sodium gluconate (CeHnNaOy); sodium acetate trihydrate, (C 2 H3Na0 2 *3H 2 0); potassium chloride, (KC1); and magnesium chloride, (MgCl 2 *6H 2 0) and human serum albumin.
  • each 100 ml of multiple electrolyte isotonic solution contains 526 mg of sodium chloride, (NaCl); 502 mg of sodium gluconate (CeHnNaOy); 368 mg of sodium acetate trihydrate, (C 2 H 3 Na0 2 '3H 2 0); 37 mg of potassium chloride, (KC1); and 30 mg of magnesium chloride, (MgCl 2 » 6H 2 0) and 1 to 5% of human serum albumin.
  • the pH is adjusted with sodium hydroxide to 7.4 (or 6.5 to 8.0), sterile filtered and tested for endotoxin.
  • platelet free plasma as used in the specification in the context of the invention comprises the supernatant collected from plasma that has been centrifuged at about 17610 g and then sterile filtered.
  • platelet poor plasma or "PPP” as used in the specification in the context of the invention comprises supernatant collected from whole blood that has been centrifuged at about 2720 g.
  • the isotonic medium used in the growth factor concentrate is multiple electrolyte isotonic solution, plasma, platelet free plasma, platelet poor plasma or a combination thereof.
  • the concentrate is derived from approximately 625 x lO 6 human platelets per ml.
  • the growth factor concentrate according to an embodiment of the invention is used for treatment of facial wrinkles including but not limited to nasolabial wrinkles.
  • a therapeutic composition for intra-dermal, sub-dermal or topical administration comprising the growth factor concentrate in combination with supplemental constituents including blood, saline, silver nanoparticles, hyaluronic acid, immuno-modulatory peptides, growth factors, hormones, antibiotics, monoclonal antibodies, recombinant receptors, carriers or combinations thereof.
  • the composition can be in the form of a cream, gel, aqueous solution, spray-aerosol or transdermal patch.
  • a method of treatment of nasolabial wrinkles comprising injecting a growth factor concentrate into the nasolabial wrinkles, wherein the growth factor concentrate is derived from approximately 625 x l O 6 human platelets per ml, the concentrate comprising approximately 500 to 1000 pg/ml of EGF, 10 to 20 pg/ml of VEGF, 10 to 25 pg/ml of basic b-FGF, 20000 to 30000 pg/ml of TGF- ⁇ and 60000 to 150000 pg/ml of PDGF-AB.
  • the mode of injection of the growth factor concentrate is the fanning technique.
  • a method of preparing an intra-dermally, sub-dermally or topically administrable growth factor concentrate derived from human platelets comprising the following steps:
  • snap-freezing of the suspension is carried out at a temperature of -120°C to -200°C, thawing of the frozen suspension is carried out at 25°C to 37°C, and cellular debris are separated from the thawed suspension and the resultant suspension of growth factors is diluted with an isotonic medium before sterile-filtering.
  • a method of preparing an intra-dermally, sub-dermally or topically administrable growth factor concentrate derived from human platelets comprising the following steps:
  • a fixed number of platelets is suspended in a fixed volume of plasma to obtain the required concentration of growth factors in the growth factor concentrate, snap-freezing of the suspension is carried out at a temperature of -120°C to -200°C, thawing of the frozen suspension is carried out at 25°C to 37°C, and
  • cellular debris are separated from the thawed suspension and the resultant suspension of growth factors is diluted with an isotonic medium before sterile-filtering.
  • a method of preparing an intra-dermally, sub-dermally or topically administrable growth factor concentrate derived from human platelets comprising the following steps:
  • the snap-freezing is carried out at a temperature of -120°C to -200°C;
  • the thawing is carried out at 25°C to 37°C;
  • cellular debris are separated from the thawed suspension and the resultant suspension of growth factors is diluted with the isotonic medium before sterile-filtering and lyophilized with excipients after sterile-filtering,
  • the volume of plasma does not exceed 5 ml.
  • an intra-dermally, sub- dermally or topically administrable growth factor concentrate derived from a fixed number of human platelets, the concentrate comprising the required concentration of growth factors suspended in an isotonic medium.
  • a dosage of an intra- dermally, sub-dermally or topically administrable growth factor concentrate derived from approximately 250 ⁇ ⁇ 6 to 5000 * 10 6 human platelets per ml, the concentrate comprising approximately 900 to 2000 pg/ml of EOF, 30 to 300 pg/ml of VEGF, 20 to 100 pg/ml of b-FGF, 40000 to 120000 pg/ml of TGF- ⁇ and 200000 to 600000 pg/ml of PDGF-AB suspended in an isotonic medium. More preferably, the growth factor concentrate is derived from approximately 625 x 10 6 human platelets per ml.
  • Freshly collected platelets or fresh blood can be collected from donors or even blood banks for large scale manufacturing of GFC.
  • the blood is preferably transported to the central processing laboratory at 20-24°C in a transportation box.
  • Blood can alternatively be collected from donors requiring treatment with the GFC.
  • Blood may also be collected from other mammalian species such as horse, dog, cat, buffalo, cow, sheep, goat, rodents etc. from either jugular vein or the cephalic vein or femoral vein.
  • One part of the blood sample collected is preferably routinely processed for complete blood count (CBC), and rapid infectious disease marker testing for Human Immunodeficiency Virus (HIV- 1,2), Hepatitis B virus (HBV), Hepatitis C Virus (HCV), Venereal disease research laboratory (VDRL) tests.
  • the remaining part of the blood collected is preferably sent to a class B environment clean-room for further processing to yield the GFC.
  • the clean room temperature is preferably maintained at 22°C with a relative humidity of 55 %.
  • Stability of the platelets in the whole blood was checked for different temperatures and time points and it was found that platelets in whole blood are stable between 15 to 30°C until 24 hours for the purpose of GFC preparation i.e. the growth factor levels measured using ELISA after recovering platelets at different temperatures and time points remained stable upto 24 hours.
  • the blood sample to be used for obtaining the GFC is centrifuged at 109 g to 680 g, preferably at about 382 g for 15 minutes at 22°C for isolation of platelets. After centrifugation, three layers are observed: a top layer of yellow coloured platelet rich plasma (PRP), a middle layer of white blood corpuscles (WBC) and the bottom layer of red blood corpuscles (RBC).
  • PRP yellow coloured platelet rich plasma
  • WBC white blood corpuscles
  • RBC red blood corpuscles
  • the top layer is aspirated carefully to maximize platelet yield, while ensuring that no WBCs are picked up, and placed in another sterile centrifuge tube.
  • the platelet rich plasma collected is then centrifuged at 680 to 3442 g, preferably at about 2720 g for 10 minutes.
  • PRP platelet poor plasma
  • the entire PPP is collected in a sterile centrifuge tube and stored at room temperature for later use. Platelets of any desired concentrations could be prepared in this manner which is not possible in other known devices for obtaining PRP.
  • the platelet pellet is re-suspended in the appropriate amount of isotonic medium.
  • the isotonic medium is platelet free plasma, platelet poor plasma, multiple electrolyte isotonic solution or combinations thereof.
  • the concentrated platelet pellet can be re-suspended in 1 to 10 ml of multiple electrolyte isotonic solution or platelet free plasma which is tested for ABO compatibility before use.
  • the platelets are then counted and further isotonic medium can be added so that the platelet numbers are adjusted to the count of approximately 250 to 5000 ⁇ 10 6 platelets per ml; preferably approximately 625 ⁇ 10 6 platelets per ml.
  • This platelet suspension is then subjected to a physiological activation by freezing at - 196°C.
  • the centrifuge tube containing the concentrated platelet suspension is placed in liquid nitrogen for 120 seconds and then subjected to a rapid thawing. Rapid thawing is done at 37°C for 120 seconds.
  • freeze-thaw cycle is sufficient to physiologically activate the platelets and causes lysis of the platelet membranes.
  • This freeze-thaw cryo-stimulates the platelets to release growth factors.
  • the suspension can then be mixed with 4 to 14 ml of PPP removed at the earlier step or platelet free plasma or multiple electrolyte isotonic solution and subjected to high speed centrifugation at a speed of 17610 g for 30 minutes.
  • the last centrifugation is critical to remove all the plasma membranes or membrane antigens of platelets or debris so as to obtain an acellular solution. After high speed centrifugation supernatant is aspirated and transferred to another sterile tube.
  • the GFC is amenable to lyophilisation after mixing the GFC with 2 to 10% mannitol, sucrose and/or glycine added to the GFC as bulking agent/lyoprotectant for lyophilisation.
  • the lyophilised product is sealed with a flip off cap for clinical application and is stable for more than one year at 4°C.
  • Lyophilised GFC can be reconstituted with 5-15 ml of multiple electrolyte isotonic solution.
  • Sterile water for injection can also be used as a diluent for reconstitution, with 1% human serum albumin. Blood type matched plasma can also be used as a diluent.
  • the platelet pellet is suspended in 1ml of multiple electrolyte isotonic solution and after the freeze-thaw, the thawed solution is then mixed with 9 ml of plasma which had been removed from the upper layer at the end of the second centrifugation.
  • the multiple electrolyte isotonic solution is supplemented with pharmaceutically acceptable excipients.
  • the pharmaceutically acceptable additives are selected from a group comprising of Acid Citrate Dextrose-solution A (ACD-A), Ethylenediaminetetraacetic acid (EDTA), and Citrate phosphate dextrose adenine (CPDA),
  • GFC comprises a combination of Growth Factors such as Epidermal Growth Factor (EGF), Vascular Endothelial Growth Factor (VEGF), Transforming Growth Factor-beta (TGF- ⁇ ), basic fibroblast growth factor (bFGF), Insulin like growth factor- 1 (IGF-1), Hepatocyte Growth Factor (HGF), Platelet derived growth Factor- (PDGF-AA), (PDGF-AB), (PDGF-BB); Cytokines like RANTES, Interleukin-1 beta (IL- ⁇ ⁇ ), Macrophages inhibitory protein- 1 alpha ( ⁇ - ⁇ ), GRO- alpha, ENA-78, MCP-3, NCP, IGFBP-3; Basic Proteins like Platelet factor 4 (PF-4), Endostatin, PBP, Connective tissue activating peptide(CTAP), Neutrophil activating peptide (NAP); adhesive proteins such as ECGF, Plasminogen activator inhibitior-1, Laminin-8, Fibrinogen, Fibronectin, Thro
  • the GFC product was tested for endotoxin so as to confirm that the product was safe and free from bacterial contamination.
  • the GFC product was also tested for infectious diseases like HIV, HCV, HBV, Syphilis etc and was found to be disease-free and safe for clinical applications.
  • the snap-freezing is done in liquid nitrogen or in liquid helium.
  • the thawing can be done in a sterile water bath at 37°C.
  • the isotonic medium can be multiple electrolyte isotonic solution, plasma, platelet free plasma, platelet poor plasma or a combination thereof.
  • the multiple electrolyte isotonic solution can be supplemented with pharmaceutically acceptable excipients.
  • the excipients include mannitol, sucrose, glycine or combinations thereof.
  • the cellular debris are removed from the thawed suspension by centrifuging the thawed suspension at 1 1270 g to 17610 g for 25 to 35 minutes and isolating the supernatant.
  • the platelets of step (a) of the method can be obtained by plateletpheresis, or by centrifugation of whole blood.
  • the platelets of step (a) are preferably obtained by:
  • centrifuging at least 10 ml of anticoagulated human blood at 109 g to 680 g for 5 to 20 minutes;
  • step (b) isolating the plasma-free pellet of platelets obtained at the end of step (b).
  • the centrifugation in step (a) is carried out at 382 g for 15 minutes and the centrifugation in step (b) is carried out at 2720 g for 10 minutes.
  • a method of treating dermatological conditions comprising administering topically, intra-dermally or sub-dermally the growth factor concentrate prepared by any of the methods above.
  • the growth factor concentrate prepared by any of the methods above for treatment of human dermatological conditions.
  • the conditions can include naso-labial wrinkles, facial wrinkles and undesired age- related dermatological changes.
  • nasolabial wrinkles are treated with the growth factor concentrate derived from approximately 625 ⁇ 10 6 human platelets per ml.
  • a therapeutic composition for topical, sub-dermal or intra-dermal administration comprising the growth factor concentrate prepared by any of the methods above in combination with supplemental constituents including blood, saline, silver nanoparticles, hyaluronic acid, immuno-modulatory peptides, growth factors, hormones, antibiotics, monoclonal antibodies, recombinant receptors, carriers or combinations thereof.
  • the composition can be in the form of a cream, gel, aqueous solution, spray-aerosol or transdermal patch.
  • the GFC is administered by fanning technique of injection wherein a 28 - 30 gauge needle is inserted into the skin, 0.3 ml to 0.4 ml of GFC is injected, the needle is then retracted slightly and the needle re-inserted at a different angle and again 0.3 ml to 0.4 ml of GFC is injected. This is done repeatedly and results in a famshaped pattern of injections.
  • Example 1 In order that those skilled in the art will be better able to practice the present disclosure, the following examples are given by way of illustration and not by way of limitation.
  • vacutainers Human blood was withdrawn into vacutainers after getting informed consent from the patient. About 25-35 ml blood is collected in two types of vacutainers: 25 ml in vacutainers containing ACD-A for preparing GFC and 5-10 ml blood in EDTA tubes for infectious disease marker testing and complete blood parameter testing. Minimum 25 ml of blood was taken for 5 ml of GFC preparation. Blood was transported at 15 to 30°C preferably at 22°C within 4 hours of withdrawal. The first centrifugation was done at 382 g for 15 minutes since platelet recovery at 382 g for 15 minutes is optimum with a loss of only 8-10 % platelets. Platelet loss is significantly greater at lower or higher centrifugation speeds.
  • the solution was then snap-frozen in liquid nitrogen at -196°C for 2 minutes.
  • the frozen solution was then rapidly thawed in a water bath at 37°C for 2 minutes.
  • the thawed solution was then mixed with 4 ml of PPP. This solution was then transferred to another sterile tube and subjected to high speed centrifugation at 17610 g for 30 minutes.
  • the supernatant containing the growth factors was collected and sterile filtered through a 0.22micron filter.
  • the effective concentration of platelets used for preparation of the GFC is 625 l O 6 platelets per ml.
  • Inclusion Criteria Male and female subjects were recruited in the study. Inclusion and exclusion criteria were as follows: Inclusion Criteria:
  • Grade 0 being no nasolabial wrinkles and Grade 4 being deepest nasolabial wrinkles.
  • one 5 ml dose of GFC prepared as per Example 1 was drawn into a syringe with a 28 to 30G needle. Before injection, the face was cleaned and all the make-up was removed. The nasolabial wrinkle area was cleaned and disinfected thoroughly. A topical anaesthesia cream was applied to the injection area and kept for 20 to 30 minutes to act. The point of injection was marked with a sterile marker pen. The dose was given intra-dermally in the area of the naso-labial wrinkle by the fanning-technique.
  • FIG. 2 shows the Wrinkle Severity Rating Scale (WSRS) scores of twenty study subjects, before treatment and at one month, two months and three months after treatment with the GFC as per Example 2.
  • WSRS Wrinkle Severity Rating Scale
  • the wrinkles are scored on a scale of 0-4 with Grade 0 being no nasolabial wrinkles and Grade 4 being deepest nasolabial wrinkles. Results in FIG. 2 and FIG 3 show that the nasolabial wrinkles reduce significantly after treatment with the GFC prepared by Example 1.
  • the GFC of the present invention is derived from predetermined numbers of platelets yielding proportionate amounts of growth factors which in turn serves to provide consistent clinical results.
  • Fixed dosages of GFC can be administered to patients to treat facial wrinkles. Further, less than 10% platelets are lost by the method of the invention.
  • the platelets are physiologically activated without incorporating any additional materials or chemical substances, like calcium chloride or bovine thrombin, and hence it is safe.
  • the single freeze-thaw makes the process less time consuming and more suitable for large scale production and the growth factors are not denatured as in the case of multiple freeze-thaws.
  • the present invention provides GFCs that can have standardized concentrations which can be diluted as per the requirement to make it suitable for specific clinical indications. Another benefit of the present method is that multiple doses of GFC can be prepared from a single blood draw and hence this method is cost effective and not time consuming. Further, GFC does not show. any flocculation on long-term storage for upto six months at -20°C.
  • GFC is acellular and devoid of plasma membranes or other antigenic materials, it does not elicit any immune reactions or formation of allo-antibodies.
  • GFC can optionally be made plasma free so that it can be used as a therapeutic agent without any problem of ABO incompatible plasma that may cause immune reactions.
  • GFC is also amenable to lyophilisation so that the GFC can be stored at room temperature or in a 4°C refrigerator without any degradation for more than one year.
  • lyophilized GFC can be made into a cream, gel, aqueous solution, spray-aerosol or transdermal patch.
  • GFC is a natural i.e. non- recombinant product and the method provided by the present invention for production of GFC is economical.
  • GFC shows improved clinical outcomes due to the significantly higher level of growth factors in GFC as compared to multiple freeze-thawed HPL prepared by known methods.
  • GFC also serves as a personalised therapy for patients requiring specific concentrations of GFC to be administered as the GFC can be prepared in any desired concentration.
  • GFC contains therapeutic level of growth factors that is specifically suitable for treating facial wrinkles.
  • GFC derived from the platelets contains several important growth factors, cytokines, chemokine, anti-microbial agents and basic proteins and has very good therapeutic potential for various indications including facial wrinkles.
  • the dosage of GFC administered and method of administration according to an embodiment of the present invention result in significant reduction of facial wrinkles.
  • Growth factors such as Transforming Growth Factor- ⁇ (TGF- ⁇ ) help in proliferation of fibroblasts that in-turn helps in synthesis of collagen and elastin.
  • TGF- ⁇ Transforming Growth Factor- ⁇

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Abstract

L'invention concerne un concentré de facteurs de croissance pouvant être administré par voie intra-cutanée, sous-cutanée ou topique, obtenu à partir d'approximativement 250 x 106 à 5000 x 106 plaquettes humaines par ml pour traiter des rides faciales. Le concentré comprend approximativement 900 à 2000 pg/ml de facteur de croissance épidermique (EGF), 30 à 300 pg/ml de facteur de croissance endothélial vasculaire (VEGF), 20 à 100 pg/ml de facteur de croissance de fibroblaste de base (b-FGF), 40 000 à 12 0000 pg/ml de facteur de croissance transformant β (TGF-β) et 20 0000 à 60 0000 pg/ml de facteur de croissance dérivé de plaquette AB (PDGF-AB).
PCT/IN2013/000264 2012-08-17 2013-04-22 Concentré de facteurs de croissance pour traiter des rides faciales WO2014027360A1 (fr)

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IN2404/MUM/2012 2012-08-17

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103932911A (zh) * 2014-05-05 2014-07-23 北京志健金瑞生物医药科技有限公司 一种抗皱、美白冻干速溶化妆品及其制备方法
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