EP4419211A1 - Production method of mammalian macrophage cell derived biotechnological product - Google Patents

Production method of mammalian macrophage cell derived biotechnological product

Info

Publication number
EP4419211A1
EP4419211A1 EP23825053.4A EP23825053A EP4419211A1 EP 4419211 A1 EP4419211 A1 EP 4419211A1 EP 23825053 A EP23825053 A EP 23825053A EP 4419211 A1 EP4419211 A1 EP 4419211A1
Authority
EP
European Patent Office
Prior art keywords
media
product
process step
skin
biotechnological
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.)
Pending
Application number
EP23825053.4A
Other languages
German (de)
French (fr)
Other versions
EP4419211A4 (en
Inventor
designation of the inventor has not yet been filed The
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.)
Furkan Ayaz
Original Assignee
Furkan Ayaz
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 Furkan Ayaz filed Critical Furkan Ayaz
Priority claimed from PCT/TR2023/050023 external-priority patent/WO2024144493A1/en
Publication of EP4419211A1 publication Critical patent/EP4419211A1/en
Publication of EP4419211A4 publication Critical patent/EP4419211A4/en
Pending legal-status Critical Current

Links

Classifications

    • 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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
    • A61K8/98—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution of animal origin
    • A61K8/981—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution of animal origin of mammals or bird
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00—Preparations for care of the skin
    • A61Q19/08—Anti-ageing preparations

Definitions

  • the invention is related to a method for the production of a natural , organic, biocompatible and highly ef fective value-added product derived from mammalian cells that can be used in antiaging, wound healing, skin rej uvenating, beauti fying or skin restoring creams , gels , sprays or serum products in the cosmetic sector and without the addition of any chemical agents or preservatives other than biocompatible solutions .
  • Hyaluronic acid, collagen, or di f ferent vitamins obtained by either chemical agents or extraction methods are used in cream, gel , serum, or spray formulations known in the art .
  • Chemical active ingredients cause problems in terms of biocompatibility, while other extraction-based biological products have deficiencies in terms of ef fectiveness .
  • the aim of the ongoing studies is to develop products that are non-allergenic, non-toxic, more biocompatible , highly biologically ef ficient , minimi ze chemical usage , and have a minimal impact on the environment .
  • Products derived from conditioned medium grown in various cell culture media and in which various growth factors are involved, and their use are known in the art .
  • the application titled " Skin Cream” filed as US2011294731A1 on 12 . 02 . 2010 may be referred to as an example of the current state of the art .
  • Said document is a patent file relating to a skin conditioner comprising a compos ition comprising anti-aging cosmetics for topical application, and more particularly, a cell culture medium conditioned by cells grown in two-dimensional culture .
  • the skin cream of the said patent comprises a conditioned medium produced by incubating a medium with substantially homogeneous eukaryotic cells , at least one growth factor or extract or concentrate is present in said conditioned medium, thereby providing treatment and/or prevention of skin defect .
  • the mentioned document is a patent file on a cosmetic composition containing cell culture medium, epidermal growth factor and bovine serum albumin to improve the condition of the skin .
  • the said document relates in particular to a cosmetic composition for improving the condition of the skin containing a cell culture medium, an epidermal growth factor and bovine serum albumin as active ingredients , to a method for improving the condition of the skin using the said product , and to a method for preventing or treating skin diseases .
  • the cosmetic composition can exhibit wrinkle-reducing and wound-healing ef fects with a small amount of EGF and therefore can be useful in improving the condition of the skin or in preventing or treating skin diseases .
  • the following files may also be referred to as an example of the background of the invention : the document titled "Medicinal Composition Comprising Ips Cell Culture Supernatant And Method For Producing Same , Cosmetic And Method For Producing Same , Antiaging Compos ition, Method For Inhibiting Disease Onset , Method For Treating Disease , Method For Treating Tissue Abnormality And Beauti fication Method” filed on 12 . 01 . 2016 with the application number WO2016111377A1 and "Cosmetic And Facial Regeneration Composition Derived From Potentiated Adipose Derived Cells And Supernatants Thereof” filed on 14 . 12 . 2016 with the application number US2017165194A1 .
  • a natural , organic , biocompatible and highly ef fective biotechnological product which can be used in antiaging, wound healing, skin rej uvenating or beauti fying creams , gels , sprays or serum products and production method thereof have been developed without the addition of chemical agents or preservatives .
  • the invention is related to a method for trans forming the media obtained when growing mammalian macrophage cells into a biotechnological product for the production of a natural , organic, biocompatible and highly ef fective value-added product without the addition of any chemical agents or preservatives other than biocompatible solutions .
  • the biotechnological product obtained by the production method of the invention is also within the protection scope of the invention .
  • An obj ect of the invention is to provide a production method for a biotechnological product that allows for the elimination of the disadvantages of products that can be produced by adding various chemicals to the prior art .
  • the invention in its most general form, is a method of manufacturing a biotechnological product for dermocosmetic purposes , comprising the following process steps : a ) preparation of the media, b ) growing mammalian cells until confluency, c ) collection of confluent media at separate time points , d) obtaining the final product by mixing the collected media with a material containing at least one of the group of cream, gel , spray, serum base in crude and/or lyophili zed form .
  • 10% fetal bovine serum is used to form broths in the process step a .
  • an RPMI medium containing 1 % penicillin- streptomycin is used in the process step a .
  • mammalian cells are grown at a concentration of lx! 0 6 /mL in the process step b .
  • mammalian cells are grown until 90% confluent in the process step b .
  • the media are collected in the process step c at hours 24 , 48 and/or 72 .
  • the media collected in the process step d are mixed with a material containing at least one of the group of cream, gel , spray, serum base in 5% , 10% and/or 20% by volume .
  • the method according to an embodiment of the invention comprises the following process steps ; a ) preparation of the media using RPMI media containing 10% FBS , 1 % Pen Strep, b ) growing mammalian cells until 90% confluency at a concentration of lxl O 6 /mL, c ) when 90% confluent , grown separately in 24 , 48 and 72 hour periods and the media in each time period are collected, d) using the collected media samples for obtaining the final product by mixing them with water-based cream, gel , spray and serum bases in 5 , 10 and 20% volumes in lyophili zed or crude formulations of cream, gel , spray and serum formulations .
  • media are firstly derived from mammalian macrophage cells in the method for producing the product of the invention, which media are formed using an RPMI medium (RPMI 1640 ) containing 10% Fetal Bovine Serum ( FBS ) and 1 % Penicillin-streptomycin .
  • RPMI 1640 RPMI 1640
  • FBS Fetal Bovine Serum
  • Penicillin-streptomycin RPMI 1640
  • mammalian cells in these media are grown until 90% confluent at a concentration of lxl O 6 /mL .
  • the media are collected in slices of 24 , 48 , and 72 hours when 90% confluent .
  • the collected media samples are used for obtaining the final product by mixing with waterbased creams , gels , sprays and serum bases in 5 , 10 and 20% by volume in the formulations of creams , gels , sprays and serum bases in their lyophili zed or raw form .
  • the final product is provided in the form of a water-based cream, gel , spray, or serum base .
  • the crude or lyophili zed media are combined with biocompatible solutions to produce a dermocosmetic product to obtain the final product .
  • the product obtained by the method of the invention includes , inter alia, growth signals produced by macrophages taken from mammalian cells , anti-apoptotic signals and signals to support the survival of cells .
  • Growth signals produced by macrophages from mammalian cells , anti-apoptotic signals growth factors , signals to support cell survival , and molecules enable other cells in the environment ( as detected in epithelium and fibroblast cells ) to grow, proli ferate , and repair .
  • the mixture obtained from the cells is used as a biotechnological product in cream, gel , spray and serum formulations for wound repair, antiaging and beauty purposes .
  • the product obtained by the method of the invention comprises each of the following in concentrations ranging from 100 pg/mL to lOOOpg/mL at natural production rates: platelet-derived growth factor (PDGF) , macrophage-derived growth factor (MDGF) , bone morphogenetic proteins (BMP 2, 6, 7) family members, transforming growth factor p family members, endothelial growth factor (EGF) family members, insulin-like growth factor (IGF) family members, members of the vascular endothelial growth factor (VEGF) family, fibroblast growth factor (EGF) family members .
  • PDGF platelet-derived growth factor
  • MDGF macrophage-derived growth factor
  • BMP 2, 6, 7 bone morphogenetic proteins
  • EGF endothelial growth factor
  • IGF insulin-like growth factor
  • VEGF vascular endothelial growth factor
  • EGF vascular endothelial growth factor
  • biocompatible solutions preferably include, but are not limited to, Hank's balanced salt solution (HBSS) , phosphate buffered salt solution (PBS) .
  • HBSS Hank's balanced salt solution
  • PBS phosphate buffered salt solution
  • the product obtained by the method according to an embodiment of the invention does not contain materials with low efficiency such as collagen or urea. Thus, the toxic or allergic effects of the product are reduced.
  • mammalian macrophage cell-based biotechnological products (or active ingredients) of the invention are biocompatible and have high biological functions, their effectiveness is also high.
  • the method of the invention provides a product with a long shelf life is obtained without using a protective product.
  • the product obtained by the method according to the invention is produced in the laboratory through mammalian cells and therefore, unlike the methods that cause a decrease in the population of plants or living things, the method of the invention offers an environmentally friendly production.
  • the production method of the invention is illustrated by nonlimiting examples , which are provided only for a better understanding of the subj ect matter, and it is clear that modi fications can be made by those skilled in the art without departing from the scope of the claims .

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • Animal Behavior & Ethology (AREA)
  • Dermatology (AREA)
  • Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Gerontology & Geriatric Medicine (AREA)
  • Zoology (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Birds (AREA)
  • General Chemical & Material Sciences (AREA)
  • Epidemiology (AREA)
  • Medicinal Chemistry (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Organic Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)

Abstract

The invention relates to a method for producing biotechnological products derived within mammalian cells that can be used in anti-aging, wound healing, skin rejuvenating, beautifying or skin restoring creams, gels, sprays or serum products falling under the cosmetic sector. More particularly, the invention is related to a method for trans forming the media obtained when growing mammalian macrophage cells into a biotechnological product for the production of a natural, organic, biocompatible and highly effective value-added product without the addition of any chemical agents or preservatives other than biocompatible solutions.

Description

PRODUCTION METHOD OF MAMMALIAN MACROPHAGE CELL DERIVED BIOTECHNOLOGICAL PRODUCT
Field of the Invention
The invention is related to a method for the production of a natural , organic, biocompatible and highly ef fective value-added product derived from mammalian cells that can be used in antiaging, wound healing, skin rej uvenating, beauti fying or skin restoring creams , gels , sprays or serum products in the cosmetic sector and without the addition of any chemical agents or preservatives other than biocompatible solutions .
Background of the Invention
It is known that as a result of the loss of collagen and elastic proteins in the dermal layers and the deterioration of elasticity and skin thickness over time , fine lines and wrinkles occur on the skin and this is seen as one of the signs of aging in humans . In the art , surgical treatments such as faceli ft , laser surgery and skin peeling and interventions with inj ection treatments ( e . g . , BOTOX®) are known to prevent and/ or delay skin disorders due to said aging .
Furthermore , when the skin is damaged, for example , cut , crushed, exposed to chemicals , or deprived of oxygen, regardless of the age of the subj ect , cell death occurs in the relevant region, whereas the body initiates a sequence of chemical signals that stimulate responses aimed at healing the af fected tissues . It is well-known that the body, in response to certain situations , releases collagen to help repair damaged tissue , among other reactions . In the technical field, there are commercial products that are released for aesthetic purposes and designed to reduce , prevent or slow wrinkles on the skin, and medical products that are designed to treat tissue damage and cell loss resulting from traumatic conditions .
Hyaluronic acid, collagen, or di f ferent vitamins obtained by either chemical agents or extraction methods are used in cream, gel , serum, or spray formulations known in the art . Chemical active ingredients cause problems in terms of biocompatibility, while other extraction-based biological products have deficiencies in terms of ef fectiveness .
The aim of the ongoing studies is to develop products that are non-allergenic, non-toxic, more biocompatible , highly biologically ef ficient , minimi ze chemical usage , and have a minimal impact on the environment .
Products derived from conditioned medium grown in various cell culture media and in which various growth factors are involved, and their use are known in the art .
The application titled " Skin Cream" filed as US2011294731A1 on 12 . 02 . 2010 may be referred to as an example of the current state of the art . Said document is a patent file relating to a skin conditioner comprising a compos ition comprising anti-aging cosmetics for topical application, and more particularly, a cell culture medium conditioned by cells grown in two-dimensional culture . The skin cream of the said patent comprises a conditioned medium produced by incubating a medium with substantially homogeneous eukaryotic cells , at least one growth factor or extract or concentrate is present in said conditioned medium, thereby providing treatment and/or prevention of skin defect . Another example of the current state of the art is the document titled "Cosmetic Composition For Improving Condition Of Skin, Containing Cell Culture Medium, Epidermal Growth Factor and Bovine Serum Albumin" filed on 17 . 11 . 2015 with the number W02016080732A1 . The mentioned document is a patent file on a cosmetic composition containing cell culture medium, epidermal growth factor and bovine serum albumin to improve the condition of the skin . The said document relates in particular to a cosmetic composition for improving the condition of the skin containing a cell culture medium, an epidermal growth factor and bovine serum albumin as active ingredients , to a method for improving the condition of the skin using the said product , and to a method for preventing or treating skin diseases . It is mentioned in the said document that the cosmetic composition can exhibit wrinkle-reducing and wound-healing ef fects with a small amount of EGF and therefore can be useful in improving the condition of the skin or in preventing or treating skin diseases .
The following files may also be referred to as an example of the background of the invention : the document titled "Medicinal Composition Comprising Ips Cell Culture Supernatant And Method For Producing Same , Cosmetic And Method For Producing Same , Antiaging Compos ition, Method For Inhibiting Disease Onset , Method For Treating Disease , Method For Treating Tissue Abnormality And Beauti fication Method" filed on 12 . 01 . 2016 with the application number WO2016111377A1 and "Cosmetic And Facial Regeneration Composition Derived From Potentiated Adipose Derived Cells And Supernatants Thereof" filed on 14 . 12 . 2016 with the application number US2017165194A1 .
As an alternative to the products mentioned in the above-listed documents , a natural , organic , biocompatible and highly ef fective biotechnological product , which can be used in antiaging, wound healing, skin rej uvenating or beauti fying creams , gels , sprays or serum products and production method thereof have been developed without the addition of chemical agents or preservatives .
Detailed Description of the Invention
The invention is related to a method for trans forming the media obtained when growing mammalian macrophage cells into a biotechnological product for the production of a natural , organic, biocompatible and highly ef fective value-added product without the addition of any chemical agents or preservatives other than biocompatible solutions . The biotechnological product obtained by the production method of the invention is also within the protection scope of the invention .
An obj ect of the invention is to provide a production method for a biotechnological product that allows for the elimination of the disadvantages of products that can be produced by adding various chemicals to the prior art .
The invention, in its most general form, is a method of manufacturing a biotechnological product for dermocosmetic purposes , comprising the following process steps : a ) preparation of the media, b ) growing mammalian cells until confluency, c ) collection of confluent media at separate time points , d) obtaining the final product by mixing the collected media with a material containing at least one of the group of cream, gel , spray, serum base in crude and/or lyophili zed form .
In an embodiment of the invention, 10% fetal bovine serum is used to form broths in the process step a . In an embodiment of the invention, an RPMI medium containing 1 % penicillin- streptomycin is used in the process step a .
In an embodiment of the invention, mammalian cells are grown at a concentration of lx! 06/mL in the process step b .
In an embodiment of the invention, mammalian cells are grown until 90% confluent in the process step b .
In an embodiment of the invention, the media are collected in the process step c at hours 24 , 48 and/or 72 .
In one embodiment of the invention, the media collected in the process step d are mixed with a material containing at least one of the group of cream, gel , spray, serum base in 5% , 10% and/or 20% by volume .
In more detail , the method according to an embodiment of the invention comprises the following process steps ; a ) preparation of the media using RPMI media containing 10% FBS , 1 % Pen Strep, b ) growing mammalian cells until 90% confluency at a concentration of lxl O6/mL, c ) when 90% confluent , grown separately in 24 , 48 and 72 hour periods and the media in each time period are collected, d) using the collected media samples for obtaining the final product by mixing them with water-based cream, gel , spray and serum bases in 5 , 10 and 20% volumes in lyophili zed or crude formulations of cream, gel , spray and serum formulations .
In an embodiment of the invention, media are firstly derived from mammalian macrophage cells in the method for producing the product of the invention, which media are formed using an RPMI medium (RPMI 1640 ) containing 10% Fetal Bovine Serum ( FBS ) and 1 % Penicillin-streptomycin .
In an embodiment of the invention, mammalian cells in these media are grown until 90% confluent at a concentration of lxl O6/mL .
In an embodiment of the invention, the media are collected in slices of 24 , 48 , and 72 hours when 90% confluent .
In one embodiment of the invention, the collected media samples are used for obtaining the final product by mixing with waterbased creams , gels , sprays and serum bases in 5 , 10 and 20% by volume in the formulations of creams , gels , sprays and serum bases in their lyophili zed or raw form .
In one embodiment of the invention, the final product is provided in the form of a water-based cream, gel , spray, or serum base .
In an embodiment of the invention, the crude or lyophili zed media are combined with biocompatible solutions to produce a dermocosmetic product to obtain the final product .
The product obtained by the method of the invention includes , inter alia, growth signals produced by macrophages taken from mammalian cells , anti-apoptotic signals and signals to support the survival of cells . Growth signals produced by macrophages from mammalian cells , anti-apoptotic signals growth factors , signals to support cell survival , and molecules enable other cells in the environment ( as detected in epithelium and fibroblast cells ) to grow, proli ferate , and repair . The mixture obtained from the cells is used as a biotechnological product in cream, gel , spray and serum formulations for wound repair, antiaging and beauty purposes . The product obtained by the method of the invention comprises each of the following in concentrations ranging from 100 pg/mL to lOOOpg/mL at natural production rates: platelet-derived growth factor (PDGF) , macrophage-derived growth factor (MDGF) , bone morphogenetic proteins (BMP 2, 6, 7) family members, transforming growth factor p family members, endothelial growth factor (EGF) family members, insulin-like growth factor (IGF) family members, members of the vascular endothelial growth factor (VEGF) family, fibroblast growth factor (EGF) family members .
The product obtained by the method according to an embodiment of the invention does not contain any chemical agents or preservatives other than biocompatible solutions in cream, spray, gel and serum bases. Said biocompatible solutions preferably include, but are not limited to, Hank's balanced salt solution (HBSS) , phosphate buffered salt solution (PBS) .
The product obtained by the method according to an embodiment of the invention does not contain materials with low efficiency such as collagen or urea. Thus, the toxic or allergic effects of the product are reduced.
Since the mammalian macrophage cell-based biotechnological products (or active ingredients) of the invention are biocompatible and have high biological functions, their effectiveness is also high.
The method of the invention provides a product with a long shelf life is obtained without using a protective product. In addition, the product obtained by the method according to the invention is produced in the laboratory through mammalian cells and therefore, unlike the methods that cause a decrease in the population of plants or living things, the method of the invention offers an environmentally friendly production. In the present disclosure of the present invention, the production method of the invention is illustrated by nonlimiting examples , which are provided only for a better understanding of the subj ect matter, and it is clear that modi fications can be made by those skilled in the art without departing from the scope of the claims .

Claims

Claims
1. A method for manufacturing a biotechnological product for dermocosmetic purposes, comprising the following process steps : a. preparation of the media, b. growing mammalian cells until confluency, c. collection of confluent media at separate time points; and d. obtaining the final product by mixing the collected media with a material containing at least one of the group of cream, gel, spray, serum base in crude and/or lyophilized form.
2. A method of claim 1, wherein 10% fetal bovine serum is used to form media in the process step a.
3. A method according to claim 1 or 2, characterized in that an RPMI medium containing 1% penicillin-streptomycin is used in the process step a.
4. A method of claim 1, wherein the mammalian cells are grown at a concentration of lxlO6/mL in process step b.
5. A method of claim 1 or 4, wherein the mammalian cells are grown until 90% confluency in process step b.
6. A method of claim 1, wherein the media are collected in 24, 48 and/or 72 hours in process step c.
7. A method of claim 1, wherein the collected media are mixed in
5%, 10% and/or 20% by volume with a material containing at least one of the group of cream, gel, spray, serum base in the process step d.
8. The product produced by a method according to any of the preceding claims.
EP23825053.4A 2022-12-31 2023-01-11 METHOD FOR PRODUCING A BIOTECHNOLOGICAL PRODUCT FROM MAMMAL MACROPHAGEN CELLS Pending EP4419211A4 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
TR2023022038 2022-12-31
PCT/TR2023/050023 WO2024144493A1 (en) 2022-12-31 2023-01-11 Production method of mammalian macrophage cell derived biotechnological product

Publications (2)

Publication Number Publication Date
EP4419211A1 true EP4419211A1 (en) 2024-08-28
EP4419211A4 EP4419211A4 (en) 2025-11-19

Family

ID=92075779

Family Applications (1)

Application Number Title Priority Date Filing Date
EP23825053.4A Pending EP4419211A4 (en) 2022-12-31 2023-01-11 METHOD FOR PRODUCING A BIOTECHNOLOGICAL PRODUCT FROM MAMMAL MACROPHAGEN CELLS

Country Status (1)

Country Link
EP (1) EP4419211A4 (en)

Family Cites Families (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6372494B1 (en) * 1999-05-14 2002-04-16 Advanced Tissue Sciences, Inc. Methods of making conditioned cell culture medium compositions
US9119974B2 (en) * 2011-03-04 2015-09-01 Ahmed H. Al-Qahtani Skin cream
KR101811912B1 (en) * 2011-03-04 2017-12-22 아메드 에이치. 알-칸타니 Skin cream
US9545370B2 (en) * 2012-05-08 2017-01-17 BioRegenerative Sciences, Inc. Bioactive compositions and methods for their preparation and use

Also Published As

Publication number Publication date
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