EP4362960A1 - Mit einer lösung mit exosomen imprägniertes biologisches material - Google Patents

Mit einer lösung mit exosomen imprägniertes biologisches material

Info

Publication number
EP4362960A1
EP4362960A1 EP22740841.6A EP22740841A EP4362960A1 EP 4362960 A1 EP4362960 A1 EP 4362960A1 EP 22740841 A EP22740841 A EP 22740841A EP 4362960 A1 EP4362960 A1 EP 4362960A1
Authority
EP
European Patent Office
Prior art keywords
biological material
impregnated
exosomes
freeze
solution
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
EP22740841.6A
Other languages
English (en)
French (fr)
Inventor
Laurence Barnouin
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.)
Tbf Genie Tissulaire (tbf)
Original Assignee
Tbf Genie Tissulaire (tbf)
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 Tbf Genie Tissulaire (tbf) filed Critical Tbf Genie Tissulaire (tbf)
Publication of EP4362960A1 publication Critical patent/EP4362960A1/de
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
    • A61P17/02—Drugs for dermatological disorders for treating wounds, ulcers, burns, scars, keloids, or the like
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
    • A61K35/48—Reproductive organs
    • A61K35/51—Umbilical cord; Umbilical cord blood; Umbilical stem cells
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
    • A61K35/28—Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
    • A61K35/48—Reproductive organs
    • A61K35/50—Placenta; Placental stem cells; Amniotic fluid; Amnion; Amniotic stem cells
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2300/00—Mixtures or combinations of active ingredients, wherein at least one active ingredient is fully defined in groups A61K31/00 - A61K41/00

Definitions

  • the present invention relates to the field of biological materials impregnated with exosomes and their uses in therapeutic treatments.
  • Exosomes are a form of extracellular vesicles with a diameter of between 30 and 100 nm. They are surrounded by a lipid bilayer and float at a density of 1.13-1.19 g/ml in sucrose gradients. Vesicles with characteristics of exosomes have been isolated from multiple different body fluids, including semen, blood, saliva, breast milk, amniotic fluid, ascites fluid, cerebrospinal fluid, or bile . Over the past decade, a lot of research has been done on exosomes as a mode of intercellular communication since exosomes are secreted by various cell types including stem cells. Some studies indicate that mesenchymal stem cells secrete exosomes in vitro and that these exosomes possess therapeutic properties.
  • Exosomes contain many proteins or nucleic acids of interest such as growth factors, cytokines, heat shock proteins (HSP), amino acids, nucleic acids (DNA, RNA), metabolites, enzymes etc They also include membrane proteins that can act as receptors/ligands in different cell signaling pathways.
  • patent application US2020/397945 discloses a formulation comprising a biological material in hydrated form, including Wharton's jelly, and exosomes derived from mesenchymal stem cells, for administration in the treatment of cardiac structural damage. Said formulation is administered by catheter, injection or via a prosthesis. In this application, the biological material being simply juxtaposed with the exosomes, it therefore does not allow the capture and sustained release of said exosomes.
  • Patent application US2018/228848 discloses a biological composition comprising a mixture of non-cellular compounds, in particular exosomes, derived from placental tissues for therapeutic administration.
  • the method according to this application does not, however, make it possible to obtain a means of administering exosomes which were not present in the starting placental tissue. It also does not make it possible to have a means of administration with prolonged release of the exosomes.
  • Patent application WO2020/231702 discloses a composition comprising differentiated cells, an adhesive material in hydrated form and exosomes which may come from mesenchymal cells. In this application, the exosomes are retained on the surface of the adhesive material and not impregnated inside it. The composition must then be formulated with pharmaceutical and/or cosmetic excipients before application.
  • Patent application WO2017/140914 filed by the applicant discloses a process for preparing an allograft material forming a viro-inactivated, lyophilized and sterile membrane derived from mammalian placental tissue. It does not disclose materials impregnated with a solution comprising exosomes or means of administering them.
  • the impregnation process developed by the applicant has the advantage of also allowing the impregnation of decellularized or devitalized tissues. Due to the low porosity of the devitalized tissues, the impregnation of these tissues constitutes a real challenge as described by DUBUS M, et al. (Antibacterial and Immunomodulatory Properties of Acellular Wharton's Jelly Matrix. Biomedicines vol. 10.2 227. 21 Jan. 2022).
  • the biological material impregnated with a solution comprising exosomes according to the invention therefore makes it possible to have a form of administration of the exosomes which can be used in therapy.
  • This impregnated biological material also exhibits novel exosome release kinetics, making said device particularly interesting for therapeutic applications.
  • the impregnated biological material developed by the applicant constitutes a reservoir of exosomes allowing a gradual and prolonged release of said exosomes after administration.
  • the impregnated biological material according to the present invention also has the advantage of being able to be used directly in therapy, that is to say without requiring an additional formulation or preparation step.
  • the applicant has in particular succeeded in impregnating with exosomes materials derived from the placenta and/or derived from the umbilical cord, in particular the amniotic membrane or Wharton's jelly, which themselves possess biological properties of interest.
  • the chorioamniotic membrane which separates the fetus from the mother's endometrium in mammals, includes the amniotic membrane, or amnion, and the chorionic membrane, or chorion. These two membranes are connected by a spongy tissue membrane, also called the spongy layer, made up of collagen and proteoglycans among other things, the tissue membrane spongy with protein bridges, attached on either side to the amnion on the one hand, and to the chorion, on the other.
  • a spongy tissue membrane also called the spongy layer, made up of collagen and proteoglycans among other things, the tissue membrane spongy with protein bridges, attached on either side to the amnion on the one hand, and to the chorion, on the other.
  • the amniotic membrane is the innermost layer of the chorioamniotic membrane. Its role is to protect the fetus and keep the amniotic fluid around it.
  • This amniotic membrane is a very thin, transparent tissue that has several layers, namely an epithelial layer, a basal membrane, a compact layer and a fibroblast layer.
  • Non-vascularized and non-innervated, the amniotic membrane is rich in collagen and in various growth factors, and has properties that contribute to the healing process.
  • amniotic membrane obtained from the placenta after childbirth, is a tissue that has been used for more than a hundred years in the treatment of burns and wounds. Indeed, as early as 1910, Davies was using fetal membranes on both burns and ulcerated tissue. Trelford and Trelford-Sauder report that in 1935 authors published clinical applications of the amniotic membrane in vaginoplasty, conjunctival reconstruction, treatment of burns or wounds, and treatments relating to intra-abdominal adhesion. Trelford et al. also report that in 1952 Douglas used amnion to treat extensive injuries.
  • the stromal layer of the amniotic membrane comprising the compact layer and the fibroblastic layer, allowed a greater adhesion of the graft, and therefore its efficiency.
  • the work of Trelford et al. confirmed this fact.
  • Gindraux et al. report that from 1972, and especially since its rediscovery in 1995, other authors confirmed all the clinical applications previously presented, and also reported new indications such as the genitourinary tract, stomach, larynx , oral cavity, head and neck, whether in clinical trials or case reports.
  • the amniotic membrane is particularly rich in growth factors of the EGF (Epidermal Growth Factor), TGF (Transforming Growth Factor) and KGF (Keratinocyte Growth Factor) type, as well as in hyaluronic acid.
  • EGF Epidermal Growth Factor
  • TGF Transforming Growth Factor
  • KGF Keratinocyte Growth Factor
  • Wharton's jelly is a gelatinous connective tissue surrounding the vein and the two arteries of the umbilical cord of mammals.
  • Wharton's jelly is a substance particularly rich in constituent elements of the extracellular matrix of connective tissues, in particular in glycosaminoglycans and proteoglycans; as well as collagen fibers (types I, III, IV and V).
  • Wharton's jelly also includes many growth factors such as, but not limited to, fibroblast growth factors (FGF), insulin-like growth factors I (IGF-I), transforming growth factors (TGF) , platelet-derived growth factors (PDGFs) and epidermal growth factors (EGFs).
  • FGF fibroblast growth factors
  • IGF-I insulin-like growth factors I
  • TGF transforming growth factors
  • PDGFs platelet-derived growth factors
  • EGFs epidermal growth factors
  • Wharton's jelly can be used for their properties of aiding the healing of lesions, in particular skin lesions or ocular surface lesions. Indeed, these constituent elements of Wharton's jelly participate in the improvement of the biological processes of healing and reduction of inflammation in a patient.
  • the biological materials derived from the umbilical cord or from the placenta can serve as ideal matrices for the impregnation of active principles which will then be released during the use of these tissues for various therapeutic treatments.
  • the present invention relates to a process for obtaining an impregnated biological material comprising the steps of: a) providing a lyophilized biological material, and a solution comprising exosomes, b) bringing the biological material freeze-dried with the solution comprising exosomes, c) A biological material impregnated according to the invention is obtained.
  • freeze-dried biological material is as defined below.
  • the solution comprising exosomes is as defined below.
  • the method comprises a prior step of freeze-drying a starting biological material.
  • the method comprises a prior step of culturing the cells of interest followed by a step of collecting the exosomes in the culture medium of the cells of interest.
  • the collection of the exosomes in the culture medium of the cells of interest is carried out by centrifugation.
  • the bringing into contact of step b) is carried out by depositing the solution comprising the exosomes on the surface of the freeze-dried biological material.
  • the duration of the contacting step b) is at least 15 seconds.
  • the duration of the contacting step b) is approximately 1 minute.
  • the method further comprises a step of freeze-drying the impregnated biological material after step c).
  • the method according to the invention further comprises a sterilization step after step c).
  • the method according to the invention comprises a prior step of sterilizing the starting biological material and/or the freeze-dried biological material before step a).
  • the sterilization steps are carried out by irradiation.
  • the sterilization steps are carried out by gamma irradiation by exposing the latter to gamma radiation at 25-32 kGy.
  • the method according to the invention comprises a preliminary step of viro-inactivation of the starting biological material and/or of the freeze-dried biological material before step a).
  • the freeze-dried biological material of steps a) and/or b) is virus-inactivated and/or sterile.
  • the method according to the invention further comprises, after step c), a devitalization step.
  • the method according to the invention comprises, before step a), a devitalization step.
  • the method according to the invention further comprises, before step a), a step of shaping the freeze-dried biological material.
  • the method according to the invention further comprises, after step c), a step of shaping the impregnated biological material. In one embodiment, further takes after step c), a step of shaping the impregnated biological material by molding.
  • the method according to the invention is characterized in that the freeze-dried biological material and/or the impregnated biological material is shaped into a parallelepiped, disc, cylinder, cone or sphere.
  • the method according to the invention is characterized in that the freeze-dried biological material and/or the impregnated biological material is shaped into a parallelepiped whose length and width are between 0.2 cm and 10 cm and whose height is between 0.2 cm and 1.0 cm.
  • the method according to the invention is characterized in that the freeze-dried biological material and/or the impregnated biological material is shaped into a parallelepiped whose length and width are between 0.1 cm and 10 cm and whose height is between 0.1 cm and 1.0 cm.
  • the method according to the invention is characterized in that the freeze-dried biological material and/or the impregnated biological material is shaped into a disc whose diameter is between 0.2 cm and 10, 0cm.
  • the method according to the invention is characterized in that the freeze-dried biological material and/or the impregnated biological material is shaped into a cylinder whose diameter is between 0.2 cm and 10, 0 cm and whose height is between 0.2 cm and 1.0 cm.
  • the method according to the invention is characterized in that the freeze-dried biological material and/or the impregnated biological material is shaped into a cylinder whose diameter is between 0.15 cm and 2, 0 cm and whose height is between 0.5 cm and 3 cm.
  • the method according to the invention is characterized in that the freeze-dried biological material and/or the impregnated biological material is shaped into a cylinder whose diameter is between 0.1 cm and 10, 0 cm and whose height is between 0.1 cm and 10.0 cm.
  • the method according to the invention is characterized in that the freeze-dried biological material and/or the material biological impregnated is shaped into a sphere whose diameter is between 0.2 cm and 1.0 cm.
  • the method according to the invention is characterized in that the freeze-dried biological material and/or the impregnated biological material is shaped into a sphere whose diameter is between 0.1 cm and 1 cm. 0cm.
  • the method according to the invention is characterized in that the freeze-dried biological material and/or the impregnated biological material is shaped into a cone, the diameter of the plane of which is between 0.2 cm and 1.0 cm and whose height is between 0.2 cm and 1.0 cm.
  • the method according to the invention is characterized in that the freeze-dried biological material and/or the impregnated biological material is in powder form.
  • the method according to the invention further comprises, after step c), a step of preparing the impregnated biological material in a form suitable for parenteral administration. In one embodiment, the method according to the invention further comprises, after step c), a step of preparing the impregnated biological material in a form suitable for application to the surface of the skin and/or mucosa and/or eye and/or anal fistula.
  • the method according to the invention further comprises, after step c), a step of preparing the impregnated biological material in a form suitable for application in the vitreous of the eye or by injection subconjunctival.
  • the method according to the invention further comprises, after step c), a step of preparing the impregnated biological material in a form suitable for administration in the form of an implant.
  • biological starting material means any material derived from and/or isolated from human, animal or plant tissues.
  • starting biological material will designate a material that is neither freeze-dried nor impregnated.
  • the starting biological material comprises proteoglycans.
  • the starting biological material is connective tissue comprising proteoglycans.
  • the starting biological material is a connective tissue rich in proteoglycans.
  • the starting biological material has a solid or gel consistency.
  • the starting biological material is not liquid.
  • the cells of the starting biological material are devitalized.
  • the term "devitalized cell” means a cell whose continuity of the cellular and/or nuclear membranes is altered by a physical and/or chemical process but whose cellular content, in particular DNA or RNA, is not eliminated.
  • the cells are devitalized by at least one freezing/thawing cycle.
  • the starting biological material is decellularized.
  • the size of the biological starting material according to the invention will be chosen appropriately by the person skilled in the art, in particular in the case of therapeutic use of said biological starting material, so that it has a suitable size. to the area to be treated.
  • the starting biological material is characterized in that said starting biological material is derived from one or more tissues of human origin.
  • the starting biological material is chosen from the list consisting of the amniotic membrane or a starting biological material derived from human placenta or human umbilical cord, in particular human Wharton's jelly and/or human amniotic membrane.
  • the starting biological material is characterized in that it comes from the placenta or from the umbilical cord.
  • the starting biological material comprises umbilical cord wall.
  • the starting biological material is Wharton's jelly.
  • Wharton's jelly is meant the gelatinous biological tissue present in the umbilical cord of mammals, including the vein and the two arteries naturally included within said gelatinous biological tissue have been removed.
  • the term “Wharton's jelly” may or may not be understood to include the amniotic membrane surrounding the umbilical cord Wharton's jelly.
  • the term “Wharton's jelly” is understood to not include thick collagen fibers and/or lacunae and vascular walls (villi and intervillous chambers).
  • said biological starting material in the form of Wharton's jelly is characterized in that it is obtained according to the method described in WO2019/038411.
  • the Wharton jelly is obtained according to a process comprising the steps of: a) a segment of Wharton jelly is available from which the vein and the arteries have been removed; b) a viro-inactivation treatment of the Wharton jelly segment is carried out to obtain a viro-inactivated Wharton jelly segment; c) the segment of viro-inactivated Wharton jelly is ground to obtain a homogeneous ground product of viro-inactivated Wharton jelly.
  • Wharton's jelly does not include umbilical vessels.
  • the biological starting material comprises Wharton's jelly and the amniotic membrane which surrounds it. [00085] In one embodiment, the starting biological material is an amniotic membrane and/or is derived from an amniotic membrane.
  • amniotic membrane is meant the tissue envelope which develops around the embryo, then the fetus, in mammals during pregnancy. Its role is to protect the developing organism by maintaining the amniotic fluid around it. It sticks to the second membrane which is the chorion.
  • the amniotic membrane includes the following physiological sublayers: the epithelial cell layer, the basement membrane, the compact layer, the fibroblastic layer, the spongy layer.
  • the biological starting material comprises the spongy layer of the amniotic membrane.
  • the spongy layer of the amniotic membrane has the advantage of being very rich in proteoglycans.
  • the starting biological material in the form of an amniotic membrane is characterized in that it is obtained according to the method described in WO2017/140914.
  • the starting biological material is sterile.
  • the starting biological material is virus-inactivated.
  • viral-inactivation is meant a technique which makes it possible to greatly or totally and definitively reduce the ability of viruses to act. These, defined as inactivated, lose their pathogenic and replication capacities by a decrease in their population of 4 log during residual titrations which follow one or two independent chemical steps, whether on enveloped or non-enveloped viruses, DNA or RNA.
  • the device according to the invention is characterized in that said starting biological material is viro-inactivated according to the two steps of chemical viro-inactivation of the method described in WO20 17/140914 or W02019/038411 .
  • the two steps are a chemical treatment step with an alcohol particularly effective against enveloped viruses and a chemical treatment step with a peroxide particularly effective against naked viruses.
  • the first viro-inactivating chemical treatment step is the application of a wash, or the stay of the latter in a bath, composed of a first viro-inactivating agent which is l ethanol. Washing with purified water or a stay in a bath of purified water can advantageously be carried out after this step.
  • the first viro-inactivation agent is ethanol with an alcohol content of between 50% and 80%, and preferably at 70% v/v.
  • the first viro-inactivation step is carried out by treating with 70% v/v ethanol for about 60 minutes.
  • the second step of the viro-inactivating chemical treatment is the application of a wash, or the stay of the latter in a bath, composed of a second viro-inactivating agent which is hydrogen peroxide.
  • the second viro-inactivation agent is hydrogen peroxide in a form chosen from an aqueous solution and a gas.
  • the second viro-inactivation agent is hydrogen peroxide in the form of an aqueous solution in a concentration of between 3% and 30% w/v.
  • the starting biological material is sterile.
  • the term "sterile" means a material devoid of germs naturally or because it has been sterilized.
  • the sterilization can be carried out by any method conventionally known to those skilled in the art.
  • the sterilization will be carried out by gamma irradiation.
  • the lyophilized biological material is sterilized.
  • freeze-dried biological material means a starting biological material having undergone at least one freeze-drying step and not being rehydrated or impregnated.
  • the freeze-dried biological material is sterile and/or viro-inactivated.
  • the cells of the freeze-dried biological material are devitalized.
  • the lyophilized biological material is decellularized.
  • freeze-drying is understood to mean a technique aimed at drying a product previously frozen by sublimation. More specifically, the liquid to be removed from the product is first transformed into ice by freezing; then by primary desiccation, under vacuum, the ice is sublimated; finally by a secondary desiccation, the water molecules on the surface of the product are extracted by desorption. [000113] In one embodiment, the freeze-drying is carried out under the following conditions:
  • the first freezing step being carried out at an acclimation temperature chosen so as not to damage the structural, functional and biological integrity of the biological material
  • the second freezing step being carried out at the final freezing temperature which is lower than the acclimatization temperature
  • the primary freeze-drying step being carried out by applying a vacuum at approximately 200 microbars and an ascending temperature profile;
  • the secondary freeze-drying step being carried out by applying a vacuum at about 50 microbars and a descending temperature profile.
  • the acclimatization temperature is between -5 and -20°C and the final freezing temperature is between -40 and -60°C.
  • the ascending temperature profile is advantageously a profile according to which the freeze-drying temperature is initially set at a low initial temperature and then increased towards a final primary freeze-drying temperature, in one or more intermediate ascending temperature steps.
  • the falling temperature profile is advantageously a profile according to which the freeze-drying temperature is initially set at a temperature higher than the final temperature of the primary freeze-drying step, then which is then lowered to a final secondary freeze-drying temperature higher than the temperature initial of the primary freeze-drying step.
  • the freeze-dried biological material is viro-inactivated and/or sterile.
  • Viro-inactivation treatments as well as freeze-drying destroy the membranes of exosomes which would be naturally present in the tissues serving as matrices, the environment of these matrices after these treatments is therefore conducive to being impregnated by solutions comprising exosomes, which exosomes may be from many different cell types.
  • the content of the exosomes according to the present invention is dependent on the type of cells from which they were isolated.
  • the starting biological material comprises exosomes before impregnation.
  • exosomes according to the invention designate the exosomes which were not present in the starting biological material and/or the freeze-dried biological material before impregnation with the solution comprising exosomes according to the invention.
  • exosomes according to the present invention can be isolated from different cell types of interest depending on the pathology to be treated.
  • the isolation can be carried out by any technique known to those skilled in the art.
  • the exosomes according to the present invention are isolated by centrifugation, filtration, ultrafiltration and/or immunoprecipitation of the culture medium of the cells of interest.
  • exosomes according to the present invention can originate from a single cell type or from a mixture of different cell types.
  • the cells of interest are chosen from the group consisting of macrophages, blood platelets, dendritic cells, mesenchymal stem cells, induced pluripotent stem cells, bone marrow cells, adipose tissue and /or umbilical cord and/or are purified from biological fluids.
  • the cells of interest are genetically modified cells.
  • the exosomes according to the invention come from the treated patient (autologous) or from one or more donors (allogeneic).
  • said exosomes according to the invention are derived from mesenchymal stem cells.
  • said exosomes according to the invention are derived from human mesenchymal stem cells.
  • the human mesenchymal stem cells according to the invention are obtained using a method that does not require the destruction of the embryo.
  • said mesenchymal stem cells according to the invention are derived from the umbilical cord. [000130] In one embodiment, the mesenchymal stem cells according to the invention are derived from human umbilical cord.
  • the solution comprising exosomes according to the invention comprises a therapeutically effective amount of exosomes according to the invention.
  • the term “therapeutically effective quantity of exosomes” means the quantity of exosomes according to the invention which eliminates, attenuates or relieves the symptoms for which it is administered.
  • the solution comprising exosomes according to the invention comprises an amount of at least 10 6 exosomes according to the invention.
  • the solution comprising exosomes according to the invention comprises an amount of at least 10 9 exosomes according to the invention.
  • the solution comprising exosomes according to the invention comprises an amount of at least 10 11 exosomes according to the invention.
  • the solution comprising exosomes according to the invention is an aqueous solution.
  • the solution according to the invention further comprises the culture medium of the cells of interest from which the exosomes have been extracted.
  • the solution according to the invention further comprises phosphate buffered saline.
  • the solution according to the invention further comprises another active ingredient.
  • impregnation and/or “impregnated” is meant that the solution comprising exosomes penetrates into the freeze-dried biological material and spreads therein, diffuses therein.
  • the impregnation therefore consists in introducing the solution comprising the exosomes according to the invention into the freeze-dried biological material according to the invention. Said freeze-dried biological material is therefore at least partially rehydrated and the exosomes according to the invention are embedded in the network of proteoglycans of the impregnated biological material according to the invention so that it can be freeze-dried again in order to be preserved without losing its content in exosomes according to the invention.
  • the impregnation according to the invention makes it possible to add a defined quantity of exosomes according to the invention which was not present in the biological material before impregnation.
  • the quantification of the impregnation is carried out by an immuno-enzymatic ELISA method for detecting exosomes in the impregnation medium using a biological material according to the invention before impregnation as a negative control.
  • it is an impregnation of at least 10 6 exosomes according to the invention.
  • it is an impregnation of at least 10 9 exosomes according to the invention.
  • it is an impregnation of at least 10 11 exosomes according to the invention.
  • it is an impregnation of at least 90% of the exosomes present in the solution comprising exosomes according to the invention.
  • it is an impregnation of at least 95% of the exosomes present in the solution comprising exosomes according to the invention.
  • it is an impregnation of at least 98% of the exosomes present in the solution comprising exosomes according to the invention.
  • the present invention also relates to a biological material impregnated with a solution comprising exosomes.
  • said impregnated biological material results from the impregnation of a freeze-dried biological material with a solution comprising exosomes according to the invention.
  • the impregnated biological material according to the invention has a solid or gel consistency.
  • the impregnated biological material according to the invention is not liquid.
  • the impregnated biological material comprises proteoglycans.
  • the impregnated biological material is connective tissue comprising proteoglycans.
  • the impregnated biological material is a connective tissue rich in proteoglycans.
  • the impregnated biological material according to the invention has a form suitable for parenteral administration.
  • the impregnated biological material according to the invention has a form suitable for in situ administration.
  • the impregnated biological material according to the invention has a form suitable for peritoneal administration.
  • the impregnated biological material according to the invention has a form suitable for administration in the form of an implant.
  • the impregnated biological material according to the invention has a form suitable for application to the surface of the skin and/or of a mucous membrane and/or of the eye and/or in a fistula. anal.
  • the impregnated biological material according to the invention comprises a therapeutically effective quantity of exosomes according to the invention.
  • the impregnated biological material is impregnated with a quantity of 10 6 exosomes according to the invention.
  • the impregnated biological material is impregnated with a quantity of 10 9 exosomes according to the invention.
  • the impregnated biological material is impregnated with a quantity of 10 11 exosomes according to the invention.
  • the impregnated biological material has the shape of a parallelepiped, a disk, a cylinder, a cone or a sphere, or a powder.
  • the impregnated biological material has the shape of a parallelepiped whose length and width are between 0.1 cm and 10 cm and whose height is between 0.1 cm and 1.0 cm .
  • the impregnated biological material has a parallelepiped shape whose length and width are between 0.2 cm and 10 cm and whose height is between 0.2 cm and 1.0 cm .
  • the impregnated biological material has a disc shape whose diameter is between 0.2 cm and 10.0 cm. In one embodiment, the impregnated biological material has the shape of a cylinder whose diameter is between 0.1 cm and 10.0 cm and whose height is between 0.1 cm and 10.0 cm.
  • the impregnated biological material has a cylinder shape whose diameter is between 0.2 cm and 10.0 cm and whose height is between 0.2 cm and 1.0 cm.
  • the impregnated biological material has the shape of a cylinder whose diameter is between 0.15 cm and 2.0 cm and whose height is between 0.5 cm and 3 cm.
  • the impregnated biological material has the shape of a sphere whose diameter is between 0.1 cm and 1.0 cm.
  • the impregnated biological material has the shape of a sphere whose diameter is between 0.2 cm and 1.0 cm.
  • the impregnated biological material has a cone shape whose plane diameter is between 0.2 cm and 1.0 cm and whose height is between 0.2 cm and 1.0 cm.
  • the biological material is impregnated with at least one second active ingredient.
  • the impregnated biological material according to the invention is characterized in that said at least one second active principle is chosen from the group consisting of antibiotics, antiseptics, antivirals, monoclonal antibodies, inhibitors semisynthetic metalloproteases, immunosuppressive agents, anti-inflammatories, antifungals, anti-allergics, anesthetics, or proteins immunoadhesives, agents for preventing dry eyes, alone or in combinations.
  • the at least second active principle can be an active principle which is naturally present or not in said starting biological material.
  • the impregnated biological material is characterized in that said at least one second active principle is chosen from the group consisting of antibiotics.
  • antibiotics include tetracyclines (daunomycin, tetracycline, chlorinetetracycline, oxytetracycline, etc.), glycopeptides (vancomycin, etc.), aminoglycosides (gentamycin, etc.), aminoglycosides (tobramycin, neomycin, etc.
  • fluoroquinolones (ciprofloxacin, moxifloxacin, etc.), quinolones (gatifloxacin, etc.), polypeptides (bacitracin, polymyxin, etc.), phenolics (chloramphenicol, etc.), macrolides (erythromycin, etc.), sulfonamides (sulfacetamide, sulfamethoxazole, sulfisoxazole, etc.), cephalosporins (cefradoxil, cefoxitin, etc.), and any other antibiotics.
  • the impregnated biological material is characterized in that said at least one second active principle is chosen from the group consisting of steroidal anti-inflammatories (AIS) and/or non-steroidal (NSAIDs).
  • AIS steroidal anti-inflammatories
  • NSAIDs non-steroidal
  • NSAIDs are given below: indomethacin, nepafenac, diclofenac, bromfenac, ketorolac, suprofen, etc.
  • the impregnated biological material is characterized in that said at least one second active principle is chosen from the group consisting of immunosuppressants, a non-limiting list is given below: dexamethasone, betamethasone, etc. .
  • the impregnated biological material is characterized in that said at least one second active principle is chosen from the group consisting of antivirals, a non-limiting list is given below: ganciclovir, trifluorothymidine, aciclovir , DDI, AZT, foscarnet, vidarabine, trifluorouridine, idoxuridine, ribavirin, protease inhibitors, anti cytomegalovirus agent, etc.
  • antivirals a non-limiting list is given below: ganciclovir, trifluorothymidine, aciclovir , DDI, AZT, foscarnet, vidarabine, trifluorouridine, idoxuridine, ribavirin, protease inhibitors, anti cytomegalovirus agent, etc.
  • the impregnated biological material is characterized in that said at least one second active principle is chosen from the group consisting of antifungals, a non-limiting list is given below: fluconazole, nitrofurazone, amphotericin B, ketoconazole, etc. [000186] In one embodiment, the impregnated biological material is characterized in that said at least one second active principle is chosen from the group consisting of antiallergics, a non-limiting list is given below: methapyriline, chlorpheniramine, pyrilamine , prohenpyridamine, etc.
  • the impregnated biological material is characterized in that said at least one second active principle is chosen from the group consisting of anesthetics, a non-limiting list is given below: lidocaine, mepivacaine, etc. .
  • the impregnated biological material is characterized in that said at least one second active principle is chosen from the group consisting of agents making it possible to prevent dry eyes, a non-exhaustive list is given below. after: azithromycin, cyclosporine, lubricants etc...
  • the cells of the impregnated biological material are devitalized.
  • the impregnated biological material is decellularized.
  • the impregnated biological material according to the invention is sterile and/or virus-inactivated.
  • the impregnated biological material according to the invention is freeze-dried.
  • Biological material impregnation kit comprising at least two independent means: a) A freeze-dried biological material, b) A solution comprising exosomes.
  • the impregnation kit according to the invention comprises a freeze-dried biological material, as described above.
  • the impregnation kit according to the invention comprises a solution comprising exosomes according to the invention, as described above.
  • Another object of the present invention relates to a device for administering exosomes comprising the biological material impregnated with a solution comprising exosomes according to the invention.
  • the administration device according to the invention consists of a biological material impregnated with a solution comprising exosomes according to the invention.
  • the administration device according to the invention is an administration device with sustained release of exosomes.
  • the device according to the invention allows release of exosomes for at least 4 hours from administration.
  • the device according to the invention allows release of exosomes for at least 24 hours from administration.
  • the device according to the invention allows exosomes to be released for a period of 24 to 72 hours from administration.
  • the device according to the invention allows release of exosomes for at least 72 hours from administration.
  • the device according to the invention allows exosomes to be released for a period of 4 hours to 1 week from administration.
  • the device according to the present invention is applied to the surface of the skin and/or of a mucous membrane and/or of the eye and/or in an anal fistula.
  • the device according to the invention is an implantable device.
  • the device according to the invention is eye drops.
  • Another object of the present invention relates to a biological material impregnated with a solution comprising exosomes according to the invention for its therapeutic use.
  • An embodiment of the present invention also relates to said biological material impregnated with a solution comprising exosomes according to the invention for its therapeutic use in regenerative medicine and/or for the treatment and/or prevention of the disease of Crohn's disease and/or fistulas and/or chronic bowel disease and/or breast disease graft versus host and/or bowel inflammation and/or stroke and/or osteoarthritis and/or respiratory distress syndrome and/or burns and/or cardiac myopathies and/or esophageal strictures and/or chronic heart failure and/or cancer, in particular of the colon and/or breast and/or lung and/or pancreas and/or melanomas, and/or lysomal overload, in particular Gaucher's disease and/or Fabry's disease, and/or type III mucopolysaccharidosis disease and/or San-Fillipo's disease and/or bronchopulmonary dysplasia and/or chronic renal failure and/or mucositis after chemotherapy or
  • Another object of the present invention relates to a therapeutic treatment method comprising a step of administering said biological material impregnated with a solution comprising exosomes according to the invention.
  • Another object of the present invention relates to a method of therapeutic treatment in regenerative medicine and/or for the treatment and/or prevention of Crohn's disease and/or fistulas and/or chronic bowel diseases. and/or graft versus host disease and/or bowel inflammation and/or stroke and/or osteoarthritis and/or respiratory distress syndrome and/or burns and/ or cardiac myopathies and/or esophageal stenosis and/or chronic heart failure and/or cancer, in particular of the colon and/or of the breast and/or of the lung and/or of the pancreas and/or of the melanomas, and/or lysomal storage diseases, in particular Gaucher's disease and/or Fabry's disease, and/or type III mucopolysaccharidosis disease and/or San-fillipo's disease and/or dysplasia bronchopulmonary and/or chronic renal failure and/or mucositis after chemotherapy or radiotherapy treatment ic, and/or type I diabetes and/or
  • Another object of the present invention relates to a device for administering exosomes comprising the biological material impregnated with a solution comprising exosomes according to the invention for its therapeutic use.
  • An embodiment of the present invention also relates to said device for administering exosomes comprising the biological material impregnated with a solution comprising exosomes according to the invention for its therapeutic use in regenerative medicine and/or for the treatment and/or prevention of Crohn's disease and/or fistulas and/or chronic bowel disease and/or graft versus host disease and/or bowel inflammation and/or stroke and/or osteoarthritis and/or respiratory distress syndrome and/or burns and/or cardiac myopathies and/or esophageal strictures and/or chronic heart failure and/ or cancer, in particular of the colon and/or of the breast and/or of the lung and/or of the pancreas and/or of melanomas, and/or of lysomal storage diseases, in particular Gaucher's disease and/or Fabry's disease, and/or mucopolysaccharidosis type III disease and/or ma San-fillipo's adia and/or bronchopulmonary dysplasia and/or
  • Another object of the present invention relates to a therapeutic treatment method comprising a step of administering said device for administering exosomes comprising the biological material impregnated with a solution comprising exosomes according to the invention.
  • Another object of the present invention relates to a method of therapeutic treatment in regenerative medicine and/or for the treatment and/or prevention of Crohn's disease and/or fistulas and/or chronic bowel diseases. and/or graft versus host disease and/or bowel inflammation and/or stroke and/or osteoarthritis and/or respiratory distress syndrome and/or burns and/ or cardiac myopathies and/or esophageal stenosis and/or chronic heart failure and/or cancer, in particular of the colon and/or of the breast and/or of the lung and/or of the pancreas and/or of the melanomas, and/or lysomal storage diseases, in particular Gaucher's disease and/or Fabry's disease, and/or type III mucopolysaccharidosis disease and/or San-Fillipo's disease and/or dysplasia bronchopulmonary and/or chronic renal failure and/or mucositis after chemotherapy or radiotherapy treatment ic, and / or type I diabetes and
  • Example 1 Preparation of a freeze-dried biological material according to the invention consisting of amniotic membrane:
  • a properly informed and consenting donor in accordance with the requirements of the Declaration of Helsinki donates placental tissue from childbirth.
  • a qualification upstream of the donor is systematic. This qualification involves a search for the HIV virus, hepatitis B, C, HTLV and the pale treponema bacterium responsible for syphilis.
  • the placental tissue is recovered as soon as possible in the delivery room following delivery. It can advantageously be placed in a sterile box and then frozen at a temperature of -20°C. [000216] In the laboratory, in a sterile room, the following procedure is applied: [000217] The amniotic membrane with the spongy layer is isolated from the placenta and the chorion is removed and cleaned.
  • This isolated tissue is kept dry at a temperature of -20°C or -80°C for up to two years or be directly treated
  • the amniotic membrane undergoes a series of baths providing chemical treatment of it. The purpose of this treatment is to disinfect the amniotic membrane, and in particular its viro-inactivation. Gentle agitation at approximately 30 rotations per minute (rpm) of the liquid medium is applied during each bath in order to ensure homogeneous penetration of the solvents into the tissues.
  • amniotic membrane is placed in a bath of purified water at room temperature for approximately 3 hours. This step ensures both the thawing of the physiological tissue and a first step of cell lysis by osmotic pressure.
  • amniotic membrane is then transferred into a decontaminating bath composed of 70% v/v ethanol by volume of ethanol relative to the total volume of the solution at room temperature for approximately 1 hour.
  • Washing is performed in purified water for about 15 minutes at room temperature to remove the ethanol.
  • the amniotic membrane is transferred into a bath composed of hydrogen peroxide at 30% w/v by weight of hydrogen peroxide relative to the total volume of the solution to be room temperature for about 15 minutes.
  • amniotic membrane is then transferred into a decontaminating bath composed of hydrogen peroxide at 3% w/v by weight of hydrogen peroxide relative to the total volume of the solution at room temperature for approximately 1 hour.
  • the chemical action applied to the amniotic membrane can then be neutralized with two baths comprising dilute sodium hydroxide at a pH of 8.5.
  • the neutralization baths are carried out at room temperature for about 15 minutes.
  • the amniotic membrane is transferred into two baths of saline phosphate buffer in order to ensure its physiological rebalancing.
  • the baths are carried out at room temperature for approximately 15 minutes.
  • the amniotic membrane is transferred to a final bath in purified water, at room temperature, for at least 15 minutes and up to approximately 1 hour.
  • amniotic membrane undergoes a freeze-drying treatment.
  • amniotic membrane On a stainless steel tray, the amniotic membrane is placed between two layers of meshed methylcellulose support filters to facilitate water vapor exchange.
  • the first freezing stage is carried out at a temperature of ⁇ 10° C. for 5 minutes, then at ⁇ 15° C. for 90 minutes;
  • the second freezing stage is carried out at a temperature of ⁇ 50° C. for 125 minutes;
  • a primary freeze-drying step is carried out by applying a vacuum at 200 microbars and by applying a temperature of +10° C. for 8 hours, followed by a temperature of +25° C. for 150 minutes;
  • a secondary freeze-drying step is carried out by applying a vacuum at 50 microbars and by applying a temperature of +35°C for 5 hours, followed by a temperature of +25°C for 1 hour. .
  • a final sterilization step is performed by exposing the amniotic membrane to gamma radiation at 25-32 kGrays.
  • a virus-inactivated, freeze-dried and sterilized biological material consisting of an amniotic membrane with a spongy layer is obtained.
  • Example 2 Preparation of a lyophilized biological material according to the invention consisting of a disc of Wharton jelly:
  • a qualification upstream of the donor is systematic. This qualification involves a search for the HIV virus, hepatitis B, C, HTLV and the pale treponema bacterium responsible for syphilis.
  • the umbilical cord is recovered as soon as possible in the delivery room. It is advantageously placed in a sterile box, comprising an NaCl solution at 4°C.
  • the umbilical cord is rinsed and hydrated in successive baths of purified water, with gentle stirring, for 4 hours.
  • the blood vessels of the umbilical cord segment are identified and separated from the rest of the segment in order to retain only Wharton's jelly and the amniotic membrane which surrounds it.
  • the Wharton jelly and the amniotic membrane are used in the rest of the process under the general name of Wharton jelly, because the quantity by weight of membrane is negligible compared to the quantity by weight of Wharton jelly.
  • Wharton jelly is frozen dry at a temperature of -20°C or -80°C.
  • the Wharton jelly is thawed in the open air, at room temperature, for a period of 5 minutes. This freezing step, followed by thawing, ensures significant devitalization of the biological material.
  • Wharton's jelly undergoes a succession of baths ensuring a chemical treatment thereof.
  • the purpose of this treatment is to disinfect the Wharton jelly, and in particular its virus inactivation.
  • gentle linear agitation of the liquid medium is applied during each bath to ensure homogeneous penetration of the solvents into the tissues.
  • the Wharton jelly is placed in a bath of purified water at room temperature for approximately 3 hours. This step ensures both the end of the thawing of the physiological tissue, and a cell lysis step by osmotic pressure.
  • the Wharton jelly is transferred into a decontaminating bath composed of 70% v/v ethanol at room temperature for approximately 1 hour.
  • Washing is performed in purified water for about 15 minutes at room temperature to remove the ethanol.
  • the Wharton jelly is transferred to a bath composed of 30% w/v hydrogen peroxide at room temperature for approximately 15 minutes.
  • the Wharton jelly is transferred to a decontaminating bath composed of 3% w/v hydrogen peroxide at room temperature for approximately 1 hour.
  • the Wharton jelly obtained is virus-inactivated.
  • the chemical action applied to the viro-inactivated Wharton jelly is then neutralized in a bath comprising dilute sodium hydroxide around a pH of 8.5.
  • the treatment of the neutralization bath is carried out at room temperature for about 15 minutes.
  • the virus-inactivated Wharton jelly is transferred to a bath of physiological buffer (PBS), in order to ensure its physiological rebalancing.
  • PBS physiological buffer
  • the bath is carried out at room temperature for approximately 15 minutes.
  • the Wharton jelly is transferred to two successive baths in purified water, at room temperature, for at least 15 minutes and up to approximately 1 hour.
  • the Wharton jelly obtained according to this chemical treatment is a largely disinfected Wharton jelly, in particular virus-inactivated.
  • the Wharton jelly is inserted into a Retsch MM400 vibrating ball mill, equipped with a 35 mL bowl of zirconium oxide.
  • the Wharton jelly takes up a space of about 1/3 of the bowl.
  • a zirconium oxide ball with a diameter of 20 mm is added to the bowl with Wharton's jelly. Grinding for 1 minute is carried out at a frequency of 3 Hz.
  • the ball with a diameter of 20 mm is recovered, and 9 zirconium oxide balls with a diameter of 10 mm are added to the bowl.
  • a second grinding of 3 minutes is carried out at a frequency of 30 Hz.
  • a third grinding is carried out with 60 balls of 5mm for 3 minutes at a frequency of 30 Hz.
  • the substance obtained is a homogeneous gelled liquid substance.
  • the substance obtained is then distributed in a stainless steel mould. This distribution is carried out using a 2.5 mL syringe.
  • the viro-inactivated and ground Wharton jelly is placed on a stainless steel tray.
  • the assembly is transferred to a freeze-dryer, where a freezing step followed by a freeze-drying step are carried out according to the following methods:
  • the first freezing step is carried out at an acclimatization temperature chosen so as not to damage the structural, functional and biological integrity of the viro-inactivated and ground Wharton jelly; [000264] the second freezing step is carried out at the final freezing temperature which is lower than the acclimatization temperature; Lyophilization:
  • a primary freeze-drying step is carried out by applying a vacuum at about 200 micro-bars and by applying a rising temperature profile;
  • a secondary freeze-drying step is carried out by applying a vacuum at about 50 microbars and by applying a falling temperature profile.
  • the product obtained is a ground Wharton jelly, disinfected, in particular viro-inactivated and freeze-dried.
  • the product obtained is defined by a cylinder shape with a diameter of 2 cm and a height of 0.3 cm, which is conferred by the support following the freeze-drying step.
  • Each of the ground Wharton jelly, viro-inactivated, and freeze-dried thus formed is easily detached from its support and repositioned in the same support and placed in primary packaging which is a TYVEK® sachet made of PE-PET copolymer.
  • a final step of sterilizing the ground material of Wharton jelly, viro-inactivated and freeze-dried, is carried out by exposing this ground material to gamma radiation at 25-32 kGrays. All the sachets comprising the ground material obtained from the initial biological substance are treated simultaneously during this step of sterilization by gamma radiation.
  • a virus-inactivated, lyophilized and sterilized biological material consisting of a disk of Wharton jelly is obtained.
  • the biological material obtained is ground using a grinder, then passed through two successive sieves with a pore diameter less than 200 microns and 90 microns then packaged in glass bottles.
  • Example 3 Preparation of a freeze-dried biological material according to the invention consisting of an umbilical cord vessel:
  • a qualification upstream of the donor is systematic. This qualification involves a search for the HIV virus, hepatitis B, C, HTLV and the pale treponema bacterium responsible for syphilis.
  • the umbilical cord is recovered as soon as possible in the delivery room. It is advantageously placed in a sterile box, comprising an NaCl solution at +4°C.
  • the returned segment undergoes a succession of baths ensuring a chemical treatment thereof.
  • the purpose of this treatment is to disinfect the artery and Wharton's jelly, and in particular its viro-inactivation.
  • gentle linear agitation of the liquid medium is applied during each bath to ensure homogeneous penetration of the solvents into the tissues.
  • the returned segment is placed in a bath of purified water at room temperature for approximately 3 hours. This step ensures both the end of the thawing of the physiological tissue, and a cell lysis step by osmotic pressure.
  • the returned segment is transferred into a decontaminating bath composed of 70% v/v ethanol at room temperature for approximately 1 hour.
  • Washing is performed in purified water for about 15 minutes at room temperature to remove the ethanol.
  • the returned segment is transferred to a bath composed of 30% w/v hydrogen peroxide at room temperature for approximately 15 minutes.
  • the returned segment is transferred into a decontaminating bath composed of hydrogen peroxide at 3% w/v at room temperature for approximately 1 hour.
  • the segment obtained is virus-inactivated.
  • the chemical action applied to the viro-inactivated segment is then neutralized, in at least one bath comprising dilute sodium hydroxide around a pH of 8.5.
  • the treatment of at least one neutralization bath is carried out at room temperature for about 15 minutes.
  • the virus-inactivated segment is transferred into at least one bath of physiological buffer (PBS), in order to ensure its physiological rebalancing.
  • PBS physiological buffer
  • the at least one bath is carried out at room temperature for about 15 minutes.
  • virus-inactivated segment is transferred to two successive baths in purified water, at room temperature, for at least 15 minutes and up to approximately 1 hour.
  • the segment obtained according to this chemical treatment is a segment that has been largely disinfected, in particular virus-inactivated.
  • virus-inactivated segment undergoes freeze-drying.
  • a sterile PETG rigid support is inserted into the lumen of the virus-inactivated segment. The latter is placed on a stainless steel tray.
  • the assembly is transferred to a freeze-dryer, where a freezing step followed by a freeze-drying step are carried out according to the following methods:
  • the first freezing step is carried out at an acclimatization temperature chosen so as not to damage the structural, functional and biological integrity of the virus-inactivated segment;
  • the second freezing step is carried out at the final freezing temperature which is lower than the acclimatization temperature
  • a primary freeze-drying step is carried out by applying a vacuum at about 200 microbars and by applying a rising temperature profile;
  • a secondary freeze-drying step is carried out by applying a vacuum at around 50 microbars and by applying a falling temperature profile.
  • the returned segment is a disinfected segment, in particular viro-inactivated and freeze-dried.
  • a final sterilization step is performed by exposing it to gamma radiation at 25-32 kGrays.
  • a virus-inactivated, freeze-dried and sterilized biological material consisting of a Wharton's jelly is obtained in the lumen of an umbilical cord vessel.
  • Example 4 Process for obtaining exosomes from mesenchymal stem cells:
  • Example 5 Demonstration of the releasing capacity of a biological material impregnated with a solution comprising exosomes according to the invention:
  • a quantity of 150 ⁇ l of exosome concentrate obtained according to the protocol described in example 4 is deposited on the surface of a half-disc of Wharton jelly obtained according to the protocol described in example 2, the other half-disk serving as control.
  • the biological material consisting of a half-disk of Wharton's jelly impregnated with a solution comprising exosomes allowed the prolonged release of the exosomes for 4 hours in the medium.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Cell Biology (AREA)
  • Developmental Biology & Embryology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • Chemical & Material Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Animal Behavior & Ethology (AREA)
  • Immunology (AREA)
  • Engineering & Computer Science (AREA)
  • Biomedical Technology (AREA)
  • Epidemiology (AREA)
  • Virology (AREA)
  • Biotechnology (AREA)
  • Zoology (AREA)
  • Hematology (AREA)
  • Reproductive Health (AREA)
  • Organic Chemistry (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Pregnancy & Childbirth (AREA)
  • Communicable Diseases (AREA)
  • Oncology (AREA)
  • Dermatology (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)
  • Manufacturing Of Micro-Capsules (AREA)
EP22740841.6A 2021-06-30 2022-06-29 Mit einer lösung mit exosomen imprägniertes biologisches material Pending EP4362960A1 (de)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
FR2107114A FR3124696A1 (fr) 2021-06-30 2021-06-30 Materiau biologique imprégné de vesicules lipidiques
PCT/EP2022/067938 WO2023275164A1 (fr) 2021-06-30 2022-06-29 Materiau biologique impregne d'une solution comprenant des exosomes

Publications (1)

Publication Number Publication Date
EP4362960A1 true EP4362960A1 (de) 2024-05-08

Family

ID=77021617

Family Applications (1)

Application Number Title Priority Date Filing Date
EP22740841.6A Pending EP4362960A1 (de) 2021-06-30 2022-06-29 Mit einer lösung mit exosomen imprägniertes biologisches material

Country Status (5)

Country Link
US (1) US20240335479A1 (de)
EP (1) EP4362960A1 (de)
FR (1) FR3124696A1 (de)
IL (1) IL309610A (de)
WO (1) WO2023275164A1 (de)

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2024171223A1 (en) * 2023-02-16 2024-08-22 Tech Cell Innovations Pvt. Ltd. Freeze dried exosome-based composition and method of preparation thereof
WO2024192119A1 (en) * 2023-03-14 2024-09-19 Direct Biologics, Llc Wound healing compositions and methods thereof

Family Cites Families (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20200397945A1 (en) * 2011-12-16 2020-12-24 Cormatrix Cardiovascular, Inc. Biological Formulations and Methods for Treating Cardiac Tissue and Disorders
US11077147B2 (en) * 2015-07-20 2021-08-03 Vivex Biologics Group, Inc. Acellular biologic composition and method of manufacture
FR3047899A1 (fr) 2016-02-18 2017-08-25 Tbf - Genie Tissulaire Et Par Abreviation Tbf Procede de preparation d'un materiau d'allogreffe, produit obtenu, et utilisations de celui-ci
FR3070262B1 (fr) 2017-08-23 2020-10-16 Tbf Genie Tissulaire Tbf Composition comprenant de la gelee de wharton, procede de preparation et utilisations
WO2020231702A1 (en) * 2019-05-11 2020-11-19 Youngsuk Yi Compositions and methods containing exosomes
EP3980087B1 (de) 2019-06-04 2023-10-11 TBF Genie Tissulaire (TBF) Biologische linse mit einer amniotischen membran

Also Published As

Publication number Publication date
IL309610A (en) 2024-02-01
FR3124696A1 (fr) 2023-01-06
WO2023275164A1 (fr) 2023-01-05
US20240335479A1 (en) 2024-10-10

Similar Documents

Publication Publication Date Title
WO2023275164A1 (fr) Materiau biologique impregne d'une solution comprenant des exosomes
US11000553B2 (en) Placental tissue composition for for treating cardiac tissue damage
LU85582A1 (fr) Fractions d'acide hyaluronique ayant une activite pharmaceutique,procedes pour leur preparation et compositions pharmaceutiques les contenant
DK2373321T3 (en) Biovedhæftende platelet lysate and compositions thereof for the treatment of mucostitis
RU2240830C1 (ru) Раневое покрытие и способ его получения
EP3672607B1 (de) Zusammensetzung mit wharton-sulze, verfahren zu ihrer herstellung und ihre verwendung
EP1549328A2 (de) Verwendung einer humanen schafhäutchen-zusammesentzung zur prophylaxe und behandlung von erkrankungen und zuständen des auges und der haut
EP3980087B1 (de) Biologische linse mit einer amniotischen membran
JP7148987B2 (ja) 眼乾燥症候群および他の外傷を受けた非角化上皮表面を処置する組成物および方法
WO2011022389A1 (en) Compositions and methods for wound healing
Wang et al. Lollipop‐inspired multilayered drug delivery hydrogel for dual effective, long‐term, and NIR‐defined glaucoma treatment
Hu et al. Biological importance of human amniotic membrane in tissue engineering and regenerative medicine
CA2654021C (fr) Utilisation de fractions cellulaires du tissu adipeux pour la regeneration tissulaire post irradiation
WO2021205471A1 (en) Composition, injectable hydrogel and methods thereof
US20140220052A1 (en) Compositions and methods for wound healing, and for recruitment and activation of macrophages in injured tissues and in implanted biomaterials used for tissue engineering
FR3096888A1 (fr) Lentille biologique comprenant une membrane amniotique
FR2998179A1 (fr) Nanostructure a base de titanate pour la regeneration et l'ingenierie tissulaire
NL2033318B1 (en) Sterile human placental allografts and methods of making thereof
EP4209200A1 (de) Ringförmige strukturen aus vernetzter amnionmembran
EP4442316A2 (de) Sterile menschliche plazenta-allotransplantate und verfahren zur herstellung davon
US20240245731A1 (en) Micronized compositions for wound healing prepared from intact human amnion-chorion tissue having an intact intermediate spongy layer positioned there between
US11497791B1 (en) Isolated placental stem cell recruiting factors
US8865178B2 (en) Compositions and methods for wound healing
FR3137273A1 (fr) Structures annulaires constituées de membrane amniotique réticulée
US12128073B2 (en) Canine-specific therapeutic compositions and methods of use

Legal Events

Date Code Title Description
STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: UNKNOWN

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: THE INTERNATIONAL PUBLICATION HAS BEEN MADE

PUAI Public reference made under article 153(3) epc to a published international application that has entered the european phase

Free format text: ORIGINAL CODE: 0009012

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: REQUEST FOR EXAMINATION WAS MADE

17P Request for examination filed

Effective date: 20240129

AK Designated contracting states

Kind code of ref document: A1

Designated state(s): AL AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LI LT LU LV MC MK MT NL NO PL PT RO RS SE SI SK SM TR

DAV Request for validation of the european patent (deleted)
DAX Request for extension of the european patent (deleted)