Novel herbal composition

Abstract

The present disclosure concerns novel compositions, the medical use thereof and in particular use in the treatment of wounds, especially in the treatment of severe wounds and / or chronic wounds. Said medical use also concerns use for promoting tissue regeneration. The compositions disclosed herein relate to herbal compositions comprising a component of a species of the genus Pistacia, a component of a species of the genus Salvia and a biomatrix of animal-origin. Additionally, related pharmaceutical compositions are also disclosed.

Description

[0001] NOVEL HERBAL COMPOSITION

[0002] TECHNICAL FIELD

[0003] The present disclosure concerns novel compositions, the medical use thereof and in particular use in the treatment of wounds, especially in the treatment of severe wounds and / or chronic wounds. Said medical use also concerns use for promoting tissue regeneration. The compositions disclosed herein relate to herbal compositions comprising a biomatrix of animal origin. Additionally, related pharmaceutical compositions are also disclosed.

[0004] BACKGROUND

[0005] In pathology, a wound is an acute injury that damages the epidermis of the skin. Severe and / or deep wounds can encompass damage to several types of tissue, for example hypodermis and muscle tissue. To heal a wound, the body undertakes a series of actions collectively known as the wound healing process. Wounds can be caused by acute trauma, but also by repetitive stress effecting the epidermis, such as pressure leading to the formation of pressure wounds for example bedsores. The wound healing process may take long time which increases the risk of infections and complications and some wounds become chronic wounds, which cause significant suffering to affected patients.

[0006] There are various drugs in modem medicine which are used for wound healing. Additionally, different herbs in traditional medicine are also used in the treatment wounds.

[0007] However, said drugs and herbs are often not effective in the treatment of wounds, for example in the treatment of severe wounds and / or in the treatment of wounds that tend to reoccur and / or difficult to heal. Furthermore, there is no available drug or herb(s) that is effective in accelerating wound healing. Moreover, drugs available for wound healing often have adverse side effects. Thus, despite of great efforts in the field of modern medicine and the knowledge based on traditional medicine, it remains a challenge to provide novel compositions that may be useful in the treatment of wounds, especially of severe wounds and / or wounds that are difficult to heal, such as chronic wounds.

[0008] SUMMARY OF THE INVENTION

[0009] It is an object of the present disclosure is to provide compositions, preparation methods and medical uses thereof that reduce, or at least partially overcome, challenges in the prior art. It is an object of the present disclosure to provide compositions which are beneficial for promoting wound healing. An object is to provide preparation methods for such compositions. Moreover, it is an object of the present disclosure to provide compositions for use as medicaments. The present inventors have found that the compositions as described herein have an unexpected therapeutic potential in the treatment of wounds and tissue regeneration. In particular, the present invention relies on the unexpected increase and improvement of tissue regeneration, as evidenced by the re-epithelialization as shown in the appended Examples.

[0010] In particular, it is an object of the present disclosure to provide means for the treatment of severe wounds and / or chronic wounds, such as wounds that take long time to heal and / or tend to recur. It is an object to provide means for the treatment of wounds that are considered uncurable in the prior art. It is an object to provide means for the effective treatment of penetrating wounds, ulcers and / or boils. These and other objects are achieved in full, or at least in part, by aspects of the inventive concepts as disclosed herein.

[0011] The present invention is based on the surprising finding that the combination of a component of an herb from the genus Pistacia, a component of an herb from the genus Salvia and a biomatrix of animal-origin results in composition with unexpectedly beneficial effects for promoting wound healing, in particular of severe and / or chronic wounds. Thus, in a first aspect of the present disclosure there is provided a composition comprising

[0012] - a component of herb A, wherein said herb A is selected from a species of the genus Pistacia',

[0013] - a component of herb B, wherein said herb B is selected from a species of the genus Salvia', and

[0014] - a biomatrix of animal-origin.

[0015] The skilled person appreciates that a composition comprising herbal extracts of the components of said herb A and / or said herb B are expected to have beneficial properties similar to that of said components of said herb A and / or said herb B, respectively. Thus, in a first alternative aspect, there is furthermore provided a composition comprising

[0016] - an herbal extract derived from a component of herb A, wherein said herb A is selected from a species of the genus Pistacia',

[0017] - an herbal extract derived from a component of herb B, wherein said herb B is selected from a species of the genus Salvia', and

[0018] - a biomatrix of animal-origin.

[0019] The composition as described herein may consists of

[0020] - a component of herb A, wherein said herb A is selected from a species of the genus Pistacia',

[0021] - a component of herb B, wherein said herb B is selected from a species of the genus Salvia', and

[0022] - a biomatrix of animal-origin.

[0023] As described for example in appended Example 2 , the present inventors have found that the combination of a component of an herb from the genus Pistacia and a component of an herb from the genus Salvia with a biomatrix of animal-origin have a surprisingly beneficial effect in promoting healing of severe wounds. Moreover, the inventors demonstrate that said composition is effective in the treatment of chronic wounds as illustrated in appended Example 3. The inventors have found that a composition according to the disclosure exhibits antimicrobial activity, such as antibacterial activity. In line with this, it is shown in the appended Examples that no animals in the experimental model exhibited infections of the wounds. Additionally, the inventors have realized that said composition according to the disclosure exhibits anti-inflammatory activity. Moreover, the inventors have found that said composition promotes tissue regeneration. Particularly, said composition expedites the process of wound healing. As shown in the appended Examples, the administration of the composition according to the present disclosure significantly reduced the healing time of wounds compared to the control compositions. The present inventors have thus found that the composition according to the present disclosure is advantageous in each phases of wound healing, such as coagulation, inflammation, migrationproliferation and remodeling. For example, by preventing and / or counteracting inflammation, the composition as disclosed herein is useful in preventing the formation of chronic wounds. Thus, the composition as disclosed herein is advantageous in maintaining and / or accelerating the normal healing process and preventing its interruption and / or deregulation, which would otherwise lead to chronic wounds.

[0024] The inventors have found an unexpectedly large therapeutic potential of said herbal components in combination with a biomatrix of animal-origin. Said biomatrix of animal-origin is understood to be a complex material having a biological activity, such as a material having the ability to promote wound healing and / or tissue regeneration. Without being bound by theory, the present inventors consider that improved tissue regeneration, such as increased re-epithelialization, is beneficial for increasing the rate of wound healing as well as decreasing the scar formation. In particular, the present inventive composition is considered to improve the rate of healing, improve regeneration of tissue as well as decrease formation of undesired scar tissue and decrease contraction of tissue surrounding the wound. As demonstrated in the appended examples, in some embodiments, complete or nearly complete epithelialization of a wound treated by the composition of the present disclosure occurs within 10 days post-treatment, such as 13 days post-treatment.

[0025] As used herein, the term “biomatrix of animal-origin” refers to a material derived from animal tissues, cells or body fluids, wherein an animal is defined as a higher eukaryotic organism. Said term does not comprise a material derived from lower eukaryotic organisms, such as plants, fungi, protozoa and algae. Without being bound by theory, the present inventors envision that it may be beneficial that the biomatrix comprises extracellular matrix proteins, lipids, immune-modulating components, and / or angiogenic growth factors. Said biomatrix of animal-origin has been found to exhibit an unexpected potential in wound healing and / or tissue regeneration in combination with said herbal components.

[0026] As discussed above, the present inventors have found that herbs of the genus Pistacia have beneficial effects in the present composition when combined with the other components as disclosed herein. In the present Examples the herb Pistacia atlantica is used in the composition disclosed herein. The person skilled in the art will appreciate that other herbs of the genus Pistacia may be used in the context of the present invention, provided that they provide beneficial effects similar to those of Pistacia atlantica, for example similar effects on wound healing.

[0027] In one embodiment, said herb A is of the species Pistacia atlantica, also known as Pistacia atlantica, Desf. The skilled person is aware of that Pistacia atlantica has several common names and is also known as for example mastic tree, pistachio tree, Mt. Atlas mastic tree, Atlas pistachio, Atlantic pistacio, Atlantic terebinth, Cyprus turpentine tree, Persian turpentine tree, mastic plant, Baneh, Butm, Butum, Melengic, Almacigo, Chatlanqus and Kabul Chatlanqus.

[0028] Said herb A may be a subspecies of Pistacia atlantica. Non-limiting examples of such subspecies are Pistacia atlantica subsp. atlantica, Pistacia atlantica subsp. kurdica, Pistacia atlantica subsp. mutica and Pistacia atlantica subsp. cabulica. In one embodiment, said herb is of the subspecies Pistacia atlantica subsp. atlantica. In a particular embodiment, said herb is of the subspecies Pistacia atlantica subsp. kurdica.

[0029] Said herb A may be a wild herb or a cultivated herb. As such, said herb A may be a wild herb. As disclosed herein, the present inventors have found that a component of an herb of the genus Pistacia has beneficial effects in the present composition when combined with the other components. The person skilled in the art appreciates that beneficial active agent(s) of an herb may be present in several components, such as parts of the plant or exudates, of said herb.

[0030] Thus, the skilled person appreciates that any part of the herb may be useful in the present composition, provided it comprises said active agent(s). As used herein, the term “exudate” refers to a substance secreted by an herb. As it will be appreciated by the skilled person, said component or an extract of the said component of herb A may be used in the context of the present invention. Accordingly, the skilled person appreciates that one or more components of an herb may be advantageous in the composition as disclosed herein. Thus, said component of herb A may be selected from the group consisting of

[0031] - an element of the subterranean subsystem of said herb A;

[0032] - an element of the aerial subsystem of said herb A;

[0033] - an exudate of said herb A; and any combinations thereof.

[0034] In one embodiment, said component of herb A is selected from the group consisting of an element of the subterranean subsystem of said herb A; an element of the aerial subsystem of said herb A; and an exudate of said herb A.

[0035] The subterranean subsystem is to be understood for parts of an herb that are below ground level. Thus, said element of the subterranean subsystem of said herb A may be a root, such as a root selected from the group consisting of a taproot, a lateral root, root hairs and any combinations thereof.

[0036] The aerial subsystem is to be understood for parts of an herb that are above ground level. As used herein, the term “aerial subsystem” also encompasses the fruit and the seeds of an herb. Thus, said element of the aerial subsystem of said herb A may be selected from the group consisting of a stem, a leaf, a flower, a bud, an internode, a node, a fruit, a seed, a trunk, a branch, a twig and any combinations thereof. The skilled person is aware of that a leaf may consist of a blade and / or a petiole. Moreover, the skilled person appreciates that a flower is also known as a reproductive shoot, and it may consist of petals, sepals, stamens and / or carpels. The skilled person also understands that a fruit of the herb may consist of several fruit parts, such as a hull, which is also known as husk.

[0037] The skilled person appreciates that the discussion about the aerial and subterranean subsystems equally applies to herb A and herb B as defined herein and is not repeated merely for the sake of brevity.

[0038] That aerial parts and / or exudates of said herb A may be particularly beneficial for wound healing. Thus, in one embodiment, said component of herb A is selected from the group consisting of

[0039] - an element of the aerial subsystem of said herb A;

[0040] - an exudate of said herb A; and any combinations thereof.

[0041] In one embodiment, said component of herb A is selected from the group consisting of an element of the aerial subsystem of said herb A; and an exudate of said herb A.

[0042] In particular, the leaves, flowers and the fruit of said herb A are considered beneficial elements of the aerial subsystem of said herb A for wound healing. Thus, in one embodiment, said element of the aerial subsystem of said herb A is selected from the group consisting of a leaf, a flower, a fruit and any combinations thereof, such as the group consisting of a leaf, a flower and a fruit. Said component of herb A may be selected from the group consisting of a leaf, a flower, a fruit, an exudate and any combinations thereof, such as the group consisting of a leaf, a flower, a fruit and an exudate. In one embodiment, said element of the aerial subsystem of said herb A is a fruit. In particular, the hull of said fruit may exhibit advantageous properties for wound healing. Thus, in one embodiment said element may be a hull of said fruit. Furthermore, in one embodiment, said component of herb A is selected from the group consisting of a leaf, a flower, a hull, an exudate and any combinations thereof of said herb A, such as the group consisting of a leaf, a flower, a hull and an exudate of said herb A. In another embodiment, said component of herb A is an exudate of said herb A. The present inventors show that the exudate of herb A is beneficial in the compositions as described herein. Said exudate may be an exudate from an element of the subterranean subsystem, which element is a root, such as a root selected from the group consisting of a taproot, a lateral root, root hairs and any combinations thereof. Said exudate may be an exudate from an element of the aerial subsystem, which element is selected from the group consisting of a stem, a leaf, a flower, a bud, an internode, a node, a fruit, a seed, a trunk, a branch, a twig and any combinations thereof. In one embodiment, said exudate is an exudate from an element of the aerial subsystem, which element is selected from the group consisting of a trunk, a branch, a twig and any combinations thereof; such as the group consisting of a trunk, a branch and any combinations thereof; such as wherein the element is a trunk. Said exudate may be an exudate from an element selected from the group consisting of a branch and a twig. Said exudate may be an exudate from an element selected from the group consisting of a trunk and a twig. Said exudate may be an exudate from an element selected from the group consisting of a trunk and a branch. The person skilled in the art appreciates that beneficial active agents of an exudate from one element of herb A may be present in exudates from at least one other element of said herb A. Hence, the source of the exudate form herb A is not to be viewed as limiting in the present context. As it will be appreciated by the skilled person, said exudate or an extract of the said exudate of herb A may be used in the context of the present invention.

[0043] In one embodiment, said exudate is a resin. As used herein, the term “resin” refers to resin, oleoresin, gum and sap and equivalents thereof; and the skilled person appreciates that these terms are used interchangeably in the field.

[0044] In a particular embodiment, said exudate is a resin, such as turpentine gum. The skilled person appreciates that turpentine gum of Pistacia atlantica is also known for example as mastic tree gum, mastic tree resin, turpentine sap and Saghez baneh, referred to as in Persian. The inventors has found that said turpentine gum may be particularly useful for the compositions as disclosed herein. It will be appreciated that said component of herb A may be turpentine gum or an extract thereof. Thus, in one embodiment, said component of herb A is turpentine gum or an extract thereof.

[0045] Said component of herb A may be processed, such as mechanically processed. The skilled person appreciates that mechanical processing does not change the composition of an herb, or a component of the herb, however affects its structural properties. Non-limiting examples of mechanical processing include chopping and grinding. The skilled person appreciates that freezing can be considered mechanical, such a thermo-mechanical processing. In certain embodiments, said component of herb A is processed by freezing and / or grinding. In one embodiment said component of herb A is not processed chemically and / or enzymatically. In some embodiments, it may be beneficial to also treat the component of herb A by a chemical and / or enzymatic process. In another embodiment said component of herb A is processed chemically and / or enzymatically.

[0046] As discussed above, the present inventors found that herbs of the genus Salvia have beneficial effects in the present composition when combined the other components as disclosed herein. The herbs of the genus Salvia, such as Salvia mirzayanii, Salvia macrosiphon, Salvia officinalis and Salvia sharifii, are considerd particularly beneficial for wound healing in the present composition. Thus, in one embodiment, said herb B is selected from group consisting of the species Salvia mirzayanii, Salvia macrosiphon, Salvia officinalis and Salvia sharifii. In one embodiment, said herb B is of the species Salvia mirzayanii. Salvia mirzayanii also known as S. mirzayanii, Rech. F. & Esfand. The skilled person is aware of that Salvia mirzayanii has several common names and is also known as for example Morpujo (referred to as in Persian), Iranian sage, Marv-e-Talkh, Moor-e-Talkh. In particular embodiments, said herb B may be a subspecies of Salvia mirzayanii Said herb B may be a wild herb or a cultivated herb. As such, said herb B may be a wild herb. As disclosed herein, the present inventors have found that a component of an herb of the genus Salvia has beneficial effects in the present composition when combined with the other components. As discussed earlier, the person skilled in the art appreciates that beneficial active agent(s) of an herb may be present in several components, such as parts of said herb. Thus one or more components of the herb B may be advantageous in the composition as disclosed herein. Thus, said component of herb B may selected from the group consisting of

[0047] - an element of the subterranean subsystem of said herb B;

[0048] - an element of the aerial subsystem of said herb B; and any combinations thereof.

[0049] As it will be appreciated by the skilled person, said component of herb B used in the context of the present invention may be an extract thereof.

[0050] In one embodiment, said component of herb B is selected from the group consisting of an element of the subterranean subsystem of said herb B; and an element of the aerial subsystem of said herb B.

[0051] Said element of the subterranean subsystem of said herb B may be a root, such as a root selected from the group consisting of a taproot, a lateral root, root hairs and any combinations thereof. Said element of the aerial subsystem of said herb B may be selected from the group consisting of a stem, a leaf, a flower, a bud, an internode, a node, a fruit, a seed, a trunk, a branch, a twig and any combinations thereof, such as the group consisting of a stem, a leaf, a flower, a bud, an internode, a node, a fruit, a seed, a trunk, a branch and a twig.

[0052] In one embodiment, said component of herb B may be an element of the aerial subsystem of said herb B.

[0053] The present inventors envision that several parts of herb B comprise similar active agents and thus said parts could be exchanged for each other. Thus, in certain embodiments, said component of herb B is selected from the group consisting of a root, a leaf, a flower, a seed and any combinations thereof, such as the group consisting of a root, a leaf, a flower and a seed. In one embodiment, said component of herb B is selected from the group consisting of a leaf, a flower, a seed and any combinations thereof, such as the group consisting of a leaf, a flower and a seed. In one embodiment, said component of herb B is selected from the group consisting of a leaf, a flower, and any combinations thereof, such as the group consisting of a leaf and a flower. In a particular embodiment, said element of the aerial subsystem is a leaf.

[0054] Said element may be a leaf or an extract thereof as will be appreciated based on the discussion above.

[0055] Said component of herb B may be processed, such as mechanically processed. The skilled person appreciates that mechanical processing does not change the composition of an herb, or a component of the herb, however affects its structural properties and non-limiting examples of mechanical processing include chopping and grinding. Said component may be processed into a powder. The skilled person appreciates that heating can be considered mechanical, such a thermo-mechanical processing. In certain embodiments, said component of herb B is processed by drying, heating and / or grinding. In one embodiment, said component of herb B is not processed chemically and / or enzymatically. In some embodiments, it may be beneficial to also treat the component of herb B by a chemical and / or enzymatic process. In one embodiment, said component of herb B is processed chemically and / or enzymatically.

[0056] As appreciated by the person skilled in the art, the developmental stage of an herb may affect the presence and / or amount of active agents of said herb and / or of a component of said herb. Thus, an herb at a certain developmental stage may be particularly beneficial for the compositions as described herein. Accordingly, in one embodiment said herb B may be in a developmental stage selected from the group consisting of sprouting stage, seeding stage, vegetative stage, budding stage, flowering stage and ripening stage; such as the group consisting of flowering stage and ripening stage; such as is in a ripening stage. In one particular embodiment said herb B is fully ripe. Similarly, in one embodiment said herb A may be in a developmental stage selected from the group consisting of sprouting stage, seeding stage, vegetative stage, budding stage, flowering stage and ripening stage; such as the group consisting of flowering stage and ripening stage; such as is in a ripening stage. In one particular embodiment said herb A is fully ripe.

[0057] Antimicrobial and / or anti-inflammatory properties are considered beneficial for would healing.

[0058] In particular, the present inventors have found that the combination of the properties of said components of herb A and herb B is of beneficial effect for wound healing in the present composition. Said component of herb A may exhibit antimicrobial activity and / or anti-inflammatory activity. Said component of herb B may exhibit antimicrobial activity and / or anti-inflammatory activity. In certain embodiments, said component of herb A and said component of herb B both exhibit antimicrobial activity and / or anti-inflammatory activity. Said antimicrobial activity may be antibacterial activity. Said activity or activities is / are considered advantageous in the treatment of wounds. The present inventors envision that said antimicrobial and / or anti-inflammatory activity of said components together in the composition as disclosed herein is beneficial in the disinfection of wounds and / or in preventing the reoccurrence of an infection of a wound. Thus, said antimicrobial activity is considered to be particularly beneficial for in the treatment of infected wounds and / or chronic wounds as demonstrated in the appended Examples.

[0059] In particular, herb A exhibits potent antioxidant capabilities, as it comprises compounds effectively scavenge free radicals, preventing oxidative damage that can hinder the wound healing process and promoting a conducive environment for effective wound healing. Thus, the presence of a component of herb A in the composition as disclosed herein is considered beneficial for wound healing.

[0060] Herb B, in particular but not limited to leaves thereof, is considered to exhibit high concentration of bioactive compounds, particularly rosmarinic acid, luteolin and other flavonoids. Such bioactive compounds have antimicrobial, anti-inflammatory, and antioxidant properties, which are considered important for efficient wound healing while minimizing the risk of adverse infections. For the sake of clarity and avoidance of any doubt, as used herein in the context of beneficial properties of said compositions as well as of said components as disclosed herein, “antimicrobial and / or anti-inflammatory properties” and “antimicrobial and / or anti-inflammatory activities” are used interchangeably. The skilled person appreciates that the discussion about the antimicrobial and / or anti-inflammatory properties equally applies to said component of herb A, said component of herb B and said biomatrix of animalorigin in a composition according to the present invention, as well as to the compositions as such, as disclosed herein.

[0061] As discussed above, the present inventors show that a biomatrix of animalorigin in combination with a component of an herb from the genus Pistacia and a component of an herb from the genus Salvia have a surprisingly beneficial effect in promoting wound healing.

[0062] In one embodiment, said biomatrix of animal-origin is a naturally occurring biological material. As used herein, the term “naturally occurring biological material” refers to a material that exists in the nature and is obtained without applying any substantial modification thereto, for example is not produced by human intervention, such as by biotechnological processing. As used herein, it is to be understood that mechanical processing, such as thermo-mechanical processing of said biomatrix of animal-origin is not considered to be a substantial modification of said biomatrix of animal-origin.

[0063] It is envisioned that said biomatrix of animal-origin may be artificially produced, such as produced by biotechnological methods, for example using cells of animal-origin. In another embodiment, said biomatrix of animal-origin is an artificially produced biological material. As used herein, the term “artificially produced biological material” refers to a material that is (i) either a naturally occurring biological material processed by human intervention, such as by biotechnological processing; or (ii) produced based on the understanding of a naturally occurring biological material, for example by biotechnological methods and / or advanced medicinal technologies. The skilled person appreciates that an artificially produced biological material may be similarly beneficial in the compositions disclosed therein if said artificially produced biological material mimics the properties of said naturally occurring biological material.

[0064] In certain embodiments, said biomatrix of animal-origin is autologous. Autologous in the present context is to be understood that said biomatrix of animal-origin derives from a patient, wherein said patient is to be treated by a composition according to the present disclosure. In other embodiments, said biomatrix of animal-origin is donor derived.

[0065] The present inventors have shown that certain biomatrices of animal-origin in the compositions disclosed herein may be particularly beneficial for wound healing. Thus, in one particular embodiment, said biomatrix of animal-origin is an omentum and / or an equivalent thereof, such as wherein said biomatrix of animal-origin is an omentum. It is understood, that “omentum” is a large flat adipose tissue layer nestling the surface of the intra-peritoneal organs of an animal. Without being bound by theory, the present inventors envision that said biomatrix provides lipid scaffolding and endogenous angiogenic growth factors that promote fibroblast activation and vascularization. In particular, it is envisioned that the combination of said properties of the biomatrix with the components of herb A and herb B leads to significant improvement in wound healing and / or tissue regeneration.

[0066] As shown in the appended examples, the inventors have found that compositions as disclosed herein, comprising said omentum in combination with said components of herb A and herb B, are particularly advantageous in the treatment of wounds, wherein said omentum may exhibit functions beneficial in immune-regulation and tissue regeneration.

[0067] The skilled person appreciates that the omentum comprises a greater omentum and a lesser omentum, which may be both beneficial in the compositions as described herein. Thus, in one embodiment said omentum comprises the greater omentum or the lesser omentum. In another embodiment, said omentum comprises both the greater omentum and the lesser omentum. The present inventors envision that that certain components of the omentum, such as at least one component of the omentum or a selection of more components of the omentum may have similar advantageous properties in wound healing to that of the omentum. Thus, in one embodiment, said biomatrix of animal-origin comprises at least one component of an omentum, such as a selection of components of said omentum. Said biomatrix of animal-origin thus may consists of a component of said omentum or a selection of components of said omentum. The omentum may be utilized in the composition disclosed herein without any enzymatic digestion and / or chemical extraction. It may be beneficial that the omentum is mechanically fragment prior to mixing of the components of the composition. Such fragmentation may follow cryopreservation of the omentum.

[0068] In one embodiment, said biomatrix of animal-origin is derived from a mammal. Said biomatrix of animal-origin may be derived from an animal of the family Bovidae, such as of the subfamily Caprinae, such as of the tribe Caprini.

[0069] In another embodiment, said biomatrix of animal-origin is derived from an animal of the genus Ovis.

[0070] In a particular embodiment, said biomatrix of animal-origin is derived from an animal of the species Ovis aries. Said biomatrix of animal-origin may derive from Iran-Black sheep.

[0071] In another embodiment, said biomatrix of animal-origin is derived from an animal of the genus Capra.

[0072] In a particular embodiment, said biomatrix of animal-origin is derived from an animal of the species Capra aegagrus hircus. The skilled person is aware of that Capra aegagrus hircus is a homotypic synonym to Capra hircus and it is understood that Capra aegagrus hircus and Capra hircus define the same species. The skilled person is also aware that said species Capra aegagrus hircus is referred to as “boz” in Persian.

[0073] In one embodiment, said biomatrix of animal-origin is derived from an animal of the species Ovis aries or from an animal of the species Capra aegagrus hircus. In another embodiment, said biomatrix of animal-origin is derived from an animal of the family Rodentia, such as wherein said biomatrix of animal-origin is derived from an animal of the species Mus musculus and / or Rattus norvegicus. The skilled person appreciates that these animals may be beneficial animal models for wound healing.

[0074] In yet another embodiment, said biomatrix of animal-origin is derived from an animal of the order Primates. The skilled person appreciates that in certain applications, such as medical applications, it is advantageous that said biomatrix of animal-origin is derived from a human. Thus, in a particular embodiment, said biomatrix of animal-origin is derived from a human.

[0075] As discussed above, the present inventors have found that a biomatrix of animal-origin that has beneficial properties which promote wound healing and which are particularly beneficial in the compositions as disclosed herein. Nonlimiting examples of these properties are tissue regenerative properties, antimicrobial activity and / or anti-inflammatory activity. Thus in a particular embodiment, said biomatrix of animal-origin has tissue regenerative properties, antimicrobial activity and / or anti-inflammatory activity. In another embodiment, said biomatrix of animal-origin has tissue regenerative properties. In yet another embodiment, said biomatrix of animal-origin has antimicrobial activity, such as antibacterial activity. Moreover, in another embodiment, said biomatrix of animal-origin has anti-inflammatory activity. It is understood that a biomatrix of animal-origin may be particularly beneficial in a composition according to the present disclosure if said biomatrix of animalorigin have tissue regenerative properties, antimicrobial activity as well as anti-inflammatory activity.

[0076] In one embodiment, said biomatrix of animal-origin is processed, such mechanically processed. As used herein, it is to be understood that mechanical processing, such as thermo-mechanical processing of said biomatrix of animal-origin is not considered to be a substantial modification of said biomatrix of animal-origin. The skilled person appreciates that mechanical processing does not change the composition of a biomatrix of animal-origin, however affects its structural properties. Non-limiting examples of mechanical processing include chopping and grinding. The skilled person appreciates that freezing can be considered mechanical, such a thermomechanical processing. Thus in one embodiment, said biomatrix of animalorigin is processed by freezing and / or grinding.

[0077] In one embodiment said composition comprises approximately equal amounts of said component of herb A, and of said component of herb B by weight. The composition comprises approximately 1 to 2 times more of the biomatrix of animal-origin by weight than of said component of herb A, and of said component of herb B by weight. In one embodiment, the weight ratio of said component of herb A, said component of herb B and said biomatrix of animalorigin in a composition according to the disclosure is in the range of 1 :1 :1 to 1 :1 :2. In one particular embodiment, the weight ratio of said component of herb A, said component of herb B and said biomatrix of animal-origin in a composition according to the disclosure is approximately 1 :1 :1.5, respectively.

[0078] As discussed above in the case of individual components of the compositions as disclosed herein, the present inventors have found that the combination of the properties of said components of herb A and herb B together with the properties of said biomatrix of animal-origin in the present composition is of beneficial effect for wound healing. Non-limiting examples of these properties are tissue regenerative properties, antimicrobial activity and / or antiinflammatory activity. Thus in a particular embodiment, a composition as disclosed herein has tissue regenerative properties, antimicrobial activity and / or anti-inflammatory activity. In another embodiment, a composition as disclosed herein has tissue regenerative properties. In yet another embodiment, a composition as disclosed herein has antimicrobial activity, such as antibacterial activity. Moreover, in another embodiment, a composition as disclosed herein has anti-inflammatory activity. It is understood that a composition as disclosed herein may be particularly beneficial in a composition according to the present disclosure if said biomatrix of animal-origin have tissue regenerative properties, antimicrobial activity as well as anti-inflammatory activity. The properties of a composition as disclosed herein are unexpectedly advantageous in the treatment of wounds. For example, the present inventors have found that a composition as disclosed herein may be useful in the treatment of bleeding wounds, wherein said composition reduces and / or prevents bleeding. Moreover, the present inventors have found that a composition as disclosed herein is beneficial in the disinfection of wounds and / or in preventing the reoccurrence of an infection of a wound. Thus, said composition is particularly beneficial in the treatment of infected wounds and / or chronic wounds. Without being bound by theory, the inventors envision that the compositions as disclosed herein are beneficial in the treatment of infections, such as infections of the stomach and / or the intestines. Moreover, the present inventors demonstrate that said composition may be particularly effective in tissue regeneration, for example in the treatment of severe wounds, which are normally characterized by extensive tissue damage. Without being bound by theory, it is envisioned that a composition as disclosed herein is beneficial in wound healing by being efficient in the disinfection of a wound, by reducing inflammation as well as by being effective in promoting tissue regeneration of the damaged tissues. As demonstrated in the appended examples, the composition as disclosed herein leads to a reduction of inflammatory cell infiltration at the wound area, such as at least 30%, such as at least 40%, such as at least 50%, such as at least 55%, such as approximately 60%, such as at least 60%, reduction in inflammatory cell infiltration compared inflammatory cell infiltration in untreated control wounds.

[0079] The skilled person will appreciate that the embodiments discussed above in relation to the first aspect of the present disclosure, are equally relevant and applicable to the second, third and further aspects disclosed herein. This particularly applies to embodiments relating to the wound healing properties of the disclosed compositions. For the sake of brevity these will not be repeated in relation to the below aspects or will only be briefly mentioned. Thus, in a second aspect of the present disclosure, there is provided a preparation method for a composition as defined herein, wherein said method comprises a step of

[0080] (i) mixing of said component of herb A or said herbal extract thereof; said component of herb B or said herbal extract thereof; and said biomatrix of animal-origin; wherein a mixture is obtained.

[0081] The skilled person appreciates that for a preparation method of a composition which comprises several components, it may be advantageous to further process said mixture, for example by mechanical processing. Thus, in one embodiment said method further comprises a step of

[0082] (ii) grinding of the mixture obtained in step (i); wherein a ground mixture is obtained.

[0083] In another embodiment, said method further comprises a step of

[0084] (iii) cooling of said mixture obtained in step (i); or said ground mixture obtained in step (ii).

[0085] In yet another embodiment, said method further comprises a step of

[0086] (iv) grinding of said mixture obtained in step (i); or said ground mixture obtained in step (ii); or the cooled mixture obtained in step (iii); wherein a reprocessed mixture is obtained.

[0087] In one embodiment, the method comprises processing the mixture obtained in step (i) by two, or all three, of steps (ii), (iii) and (iv). It will be appreciated that the compositions of the present disclosure beneficially comprises components evenly distributed within in the compositions.

[0088] It is also appreciated, that for a preparation method of a composition which comprises several components, it may be advantageous to process the individual components of said composition prior to mixing them together. Thus, in one embodiment, prior to step (i), said method comprises at least one step selected from the group consisting of:

[0089] - freezing said component of herb A; - grinding of said component of herb A; and

[0090] - sieving of said component of herb A.

[0091] In one embodiment, prior to step (i), said method comprises

[0092] - freezing said component of herb A;

[0093] - grinding of the frozen component of herb A; and

[0094] - sieving of the ground component of herb A.

[0095] As discussed above, it should be understood that said freezing, grinding and sieving are non-limiting examples of mechanical processing, and they do not change the component of said herb A, however they affect its structural properties.

[0096] The freezing temperature may be between approximately -5 to -25 °C, such as between approximately -5 to -15 °C, such as between approximately -5 to - 10 °C, such as approximately -8 °C. Moreover, the freezing duration may be at least approximately 1 hour to 24 hours, such as at least approximately 12 hours to 36 hours, such as approximately 24 hours. The skilled person appreciates that a freezing duration may be chosen based on the freezing temperature. It is also appreciated that a freezing temperature at -5°C or below may be used. It should be understood that by the end of the freezing step, said component of herb A is frozen. It will be appreciated that a shorter freezing duration may be used if the freezing temperature is lower compared to when a higher freezing temperature is used. A benefit of freezing is that a grinding step of a component of herb A is more efficient. It is also appreciated that freezing and / or grinding of a component of herb A may be beneficial for obtaining a homogeneous mixture of said components. It is to be understood that the component may be ground in frozen state.

[0097] In one embodiment, said sieving is performed with a mesh size of approximately 1 mm or less, such as approximately 500 pm or less, such as approximately 250 pm or less, such as approximately 200 pm or less, such as approximately 150 pm or less, such as approximately 100 pm or less, such as approximately 50 pm or less. The mesh size may be a mesh size used for sieving wheat-flour. The skilled person appreciates that said sieving results in the removal of non-desired particles having a bigger size than that of the mesh size. It is furthermore appreciated that said sieving may be useful for obtaining an even particle size of said desired component of herb A. As such, said desired particle size may be a particle size that enables thorough mixing of said component of herb A with the other components of the compositions as disclosed herein. In a particular embodiment, said component of herb A after the sieving step is a powder, such as a powder having a homogeneous particle size.

[0098] As solutions, such as aqueous solutions, such as water, may affect a concentration of an active agent in a component of an herb, the skilled person appreciates that it may be beneficial to avoid the use such solutions in the above processing method of said herb A. Thus, in one embodiment, said method does not comprise a cleaning step of said component of herb A, such as a cleaning step using a solution, such as a cleaning step using an aqueous solution, such as water.

[0099] In one embodiment, prior to step (i), said method comprises

[0100] - drying said component of herb B.

[0101] The skilled person appreciates that said drying may be performed by different means. For example, said drying may be performed by heating. Alternatively, drying may be air drying with or without application of heat. As discussed above, drying and heating are to be understood non-limiting examples of mechanical processing, and they do not change the component of said herb B, however they affect its structural properties. The heating temperature may be between approximately 50 °C to 250 °C, such as between approximately 100 °C to 200 °C, such as between approximately 150 °C to 200 °C, such as between approximately 170 °C to 180 °C, such as approximately 175 °C. The heating duration may be approximately 1 min to approximately 5 min, such as approximately 1 min to approximately 4 min, such as approximately 1 min to approximately 4 min, such as approximately 2 min to approximately 4 min, such as approximately 2 min to approximately 3 min. The heating duration may be approximately 1 min. The skilled person appreciates that a heating duration may be chosen based on the heating temperature. Moreover, it is appreciated that said drying and / or heating does not deteriorate beneficial effects of said component of herb B in wound healing when used in compositions as disclosed herein. The present inventors consider that drying of a component of herb A makes it easier to obtain a mixture of the components in the processing method. Said dried component of herb B may be processed into a powder prior to step (i).

[0102] In one embodiment, prior to step (i), said method comprises

[0103] - freezing said biomatrix of animal-origin.

[0104] The freezing temperature may be between approximately -5 to -25 °C, such as between approximately -5 to -15 °C, such as between approximately -5 to - 10 °C, such as approximately -8 °C. In one embodiment, the freezing duration is at least approximately 1 hour. Moreover, the freezing duration may be at least approximately 1 hour to 24 hours, such as at least approximately 12 hours to 36 hours, such as approximately 24 hours. As discussed above, the skilled person appreciates that a freezing duration may be chosen based on the freezing temperature. It is also appreciated that a freezing temperature at -5°C or below may be used. It should be understood that by the end of the freezing step, said biomatrix of animal-origin is frozen. It will be appreciated that a shorter freezing duration may be used if the freezing temperature is lower compared to when a higher freezing temperature is used. Moreover, said freezing duration may be prolonged if said biomatrix of animal-origin is to be stored prior to further processing. It is to be understood that the biomatrix of animal-origin may be ground in frozen state as further discussed below.

[0105] For the composition as disclosed herein which comprises several components, it is beneficial to apply grinding, such as thorough grinding, in a preparation method for such composition. Thus, in one embodiment, said grinding in step (ii) and / or (iv) is performed using a mixer with a rotation of at least approximately 5000 RPM, such as between approximately 5000 RPM to 25000 RPM, such as between approximately 10000 RPM to 25000 RPM, such as between approximately 15000 RPM to 20000 RPM.

[0106] It is advantageous if the mixture of said components of the compositions as disclosed herein is a homogeneous mixture, as said components are evenly distributed in said composition. Thus, in certain embodiments, the ground mixture obtained in step (ii) and / or said reprocessed mixture obtained in step (iv) is a homogeneous mixture. As used herein, the term “homogeneous mixture” refers to that said mixture is thoroughly mixed and said components are evenly distributed in said composition.

[0107] In one embodiment, the cooling temperature in step (iii) is between approximately 0 to 10 °C, such as between approximately 0 to 8 °C, such as between approximately 4 to 8 °C, such as between approximately 4 to 5 °C or such as between approximately 6 to 8 °C. Moreover, the cooling duration in step (iii) may be between approximately 1 hour to 96 hours, such as between approximately 24 hours to 96 hours, such as between approximately 48 hours to 96 hours, such as between approximately 60 hours to 80 hours, such as approximately 72 hours. Similarly, to the freezing temperature and duration as discussed above, the skilled person appreciates that a cooling duration may be chosen based on the cooling temperature. It should be understood that by the end of the cooling step, said mixture is cold. In one embodiment, said cooling duration is at least approximately 72 hours, such as approximately 72 hours. Without being bound by theory, it is envisioned that said cooling in step (iii) aids in providing the beneficial effects of a composition disclosed herein. In one embodiment, the grinding duration in step (ii) and / or (iv) is at least approximately 10 min, such as at least approximately 15 min, such as at least approximately 20 min, such as at least approximately 30 min. Moreover, the grinding duration in step (ii) and / or (iv) may be between approximately 10 min to 60 min, such as between approximately 20 min to 40 min, such as approximately 30 min. The skilled person appreciates that said grinding duration may be chosen based on the means used for said grinding, and for example may be dependent on the type of mixer used for said grinding, such as the rotation thereof.

[0108] In one embodiment of the preparation method, approximately equal amounts of said component of herb A, and of said component of herb B by weight are mixed in step (i). The composition comprises approximately 1 to 2 times more of the biomatrix of animal-origin by weight than of said component of herb A, and of said component of herb B by weight. Thus, approximately 1 to 2 times more of the biomatrix of animal-origin by weight are mixed in step (i). In one embodiment, the weight ratio of said component of herb A, said component of herb B and said biomatrix of animal-origin in a composition according to the disclosure is in the range of 1 :1 :1 to 1 :1 :2. In one particular embodiment, the weight ratio of said component of herb A, said component of herb B and said biomatrix of animal-origin mixed in step (i) is approximately 1 :1 :1.5, respectively.

[0109] In certain embodiments, the compositions as disclosed herein may be stored frozen, such as at a freezing temperature between approximately -5 to -25 °C, such as between approximately -5 to -15 °C, such as between approximately -5 to -10 °C, such as approximately -8 °C. It is appreciated that a freezing temperature at -5 °C or below may be used. In certain embodiments said compositions may be stored for between approximately 1 to approximately 12 months, or even longer. For example, in one embodiment, the composition is stored for approximately 6 months. It is considered that said storage at said freezing temperatures does not affect the beneficial properties of the composition as disclosed herein.

[0110] As discussed above, said preparation methods as disclosed herein are shown to be advantageous for obtaining compositions according to the present disclosure which are proven to be useful in wound healing.

[0111] Thus, in a related third aspect of the present disclosure, there is provided a composition prepared by the herein disclosed preparation methods.

[0112] It is appreciated that the composition described herein may be formulated as a pharmaceutical formulation.

[0113] Thus, in a fourth aspect of the present disclosure, there is provided a pharmaceutical formulation comprising a composition as described herein and at least one excipient(s), such as at least one pharmaceutically acceptable excipient(s). As used herein, the term “excipient” encompasses adjuvants, carriers, diluents, and vehicles. Furthermore, the composition may comprise at least one pharmaceutically acceptable stabilizer. Said adjuvants, carriers, diluents, vehicles and stabilizers are within the knowledge of the skilled person to make the appropriate choice thereof. The skilled person appreciates that the pharmaceutical composition may be adapted to be suitable for the selected administration route as well as desired administered dose.

[0114] The pharmaceutical formulation may be formulated for topical administration. The composition may be formulated as a paste, a hydrogel, a bioactive bandage, or a topical dressing. In another embodiment, said pharmaceutical formulation is formulated for oral administration.

[0115] The skilled person appreciates that different types wounds may require the selection of different administration routes. For example, internal wounds, such as wounds of the gastrointestinal tract, such as gastric ulcers, may require oral administration of a composition as disclosed herein. Other types of wounds, such as external wounds, such as penetrating wounds, ulcers and / or boils, which are accessible on the external surface of the body, may require topical administration of a composition as disclosed herein.

[0116] In any of these or other routes of administration, the formulation of the composition may be adapted or adjusted according to normal pharmacological procedures, comprising the effective pharmaceutical in a form suitable for the chosen route, together with suitable excipients, such as adjuvants, carriers, diluents and vehicles, conventionally used and well- known to a person skilled in the art.

[0117] Compositions of the present invention suitable for oral administration may be presented as discrete units such as capsules, cachets, pills or tablets each containing a predetermined amount of the composition as disclosed herein; for example as a powder or granules.

[0118] Typical unit dosage compositions are those containing an effective dose, as herein recited, or an appropriate fraction thereof, of a composition as herein disclosed. It should be understood that in addition to the components particularly mentioned above, the compositions of this invention may include other agents conventional in the art having regard to the type of formulation in question, for example those suitable for oral administration may include flavoring agents.

[0119] The term “pharmaceutical formulation” is used in its widest sense, encompassing all pharmaceutically applicable compositions containing at least one active substance and optional carriers, adjuvants, diluents, constituents etc. The manufacture of pharmaceutical formulations for different routes of administration falls within the capabilities of a person skilled in the art.

[0120] The terms “administration” and “mode of administration” as well as “route of administration” are also used in their widest sense. The preparation of such compositions and formulations is generally known to those skilled formulation arts and may be applied to the formulation of the composition of the present disclosure.

[0121] It should be understood that the compositions as disclosed herein may be useful as a therapeutic agents on their own rights. Without being bound by theory, it is envisioned that a direct therapeutic effect may be accomplished by the combination of said components of herb A and herb B together with said biomatrix of animal-origin.

[0122] Thus, in a fifth aspect of the present disclosure, there is provided a composition as described herein, or a pharmaceutical formulation as described herein for use as a medicament.

[0123] As explained above, the inventors demonstrate that the compositions according to the present disclosure are beneficial in promoting wound healing. It is thus envisioned that said compositions are advantageous for use in the treatment of wounds. As discussed earlier, said compositions are particularly beneficial for use in the treatment of severe wounds, infected wounds and / or chronic wounds. Non-limiting examples of wounds that may be treated by the use of the disclosed compositions include penetrating wounds, boils and / or ulcers.

[0124] As used herein, the term “wound” refers to an injury that involves lacerated or punctured skin or mucus membrane.

[0125] As used herein, the term “penetrating wound” refers to an injury that involves lacerated or punctured skin damaging the epidermis of the skin. It is to be understood that penetrating wounds encompass wounds wherein all layers of the skin are damaged, as well as wounds wherein tissue underneath the skin is also damaged. For example, such tissue underneath the skin may be adipose tissue, muscle tissue or other tissue.

[0126] As used herein, the term “boil” refers to is a deep folliculitis, which is an infection of the hair follicle and may also be referred to as a furuncle.

[0127] As used herein, the term “ulcer” refers to open sore on an external or internal surface of the body, caused by a break in the skin or mucous membrane which fails to heal. Ulcers range from small, painful sores in the mouth to bedsores and serious lesions of the stomach or intestine.

[0128] The skilled person is familiar with the terms wound, boil and ulcer.

[0129] Thus, in a sixth aspect of the present disclosure there is provided a composition as described herein, or a pharmaceutical formulation as described herein for use in the treatment of a wound. For example, said use may be for wound healing. In particular, said treatment of a wound or said wound healing may be by means of tissue re-generation, such as increased tissue regeneration. In particular, said treatment of a wound or said wound healing may be by means of re-epithelialization, such as increased re- epithelialization. For example, said use may be for promoting tissue regeneration, such as re-epithelialization of a wound.

[0130] As used herein, the term “treatment” is used in the context of therapeutic treatment and relates to the treatment, such as causative or symptomatic treatment, of a disease or disorder, the alleviation of symptoms thereof and / or prevention of said disease or disorder. For example, it is envisioned that a wound may be treated and / or alleviated. It is also envisioned that a wound, such as wound that tends to reoccur is prevented by said treatment. As discussed above, it is envisioned that the compositions and / or pharmaceutical formulations are useful in the treatment of different types of wounds. In one embodiment, said wound is a penetrating wound, a boil or an ulcer.

[0131] In another embodiment, said wound is a penetrating wound. The present inventors demonstrate that said composition is particularly beneficial in the treatment of penetrating wounds, such as severe penetrating wounds.

[0132] The person skilled in the art appreciates that the disclosed compositions may be useful in the treatment of several types of penetrating wounds. Nonlimiting examples of such wounds are surgical wounds and / or incisions, bums, bites, abrasions, lacerations and skin tears. Thus, in one embodiment, said penetrating wound is selected from the group consisting of a surgical wound and / or an incision, a bum, a bite, an abrasion, a laceration and a skin tear.

[0133] Said penetrating wound may be a bum. The skilled person appreciates that a bum may be for example a thermal bum, a chemical bum and / or an electric bum. In one embodiment, said bum is a thermal bum.

[0134] As discussed above, the person skilled in the art will appreciate that a composition as described herein may be formulated based on the type of the penetrating wound which is to be treated by using said composition via topical administration. As such it may be formulated in a shape of plaster or a bandage for example. It is to be understood that said wound is in contact with the composition during topical administration. Therefore, the dose of the composition or pharmaceutical formulation which is to be applied during topical administration may be dependent on the size, extension and / or depth of said wound.

[0135] Without being bound by theory, said properties of the compositions according to the present disclosure are considered particularly useful in the treatment of ulcers. Thus, in one embodiment, said wound is an ulcer.

[0136] The present inventors have found that the disclosed compositions are useful in the treatment of different types of ulcers. Non-limiting examples of such ulcers include peptic ulcers, mouth ulcers, arterial ulcers, venous ulcers, pressure ulcers, diabetic ulcers and genital ulcers. Thus, in one embodiment, said ulcer is selected from the group consisting of peptic ulcers, mouth ulcers, arterial ulcers, venous ulcers, pressure ulcers, diabetic ulcers and genital ulcers.

[0137] As discussed above, the person skilled in the art will appreciate that some ulcers require different administration routes than other ulcers. For example for a peptic ulcer, such as a gastric ulcer, a composition as described herein may be formulated for oral administration and administered orally. For example for a pressure ulcer, such as a bedsore, a composition as described herein may be formulated for topical administration and administered topically.

[0138] Said ulcer may be selected from the group consisting of peptic ulcers and mouth ulcers, such as gastric ulcers and / or duodenal ulcers. Said ulcer may be a gastric ulcer.

[0139] The skilled person is aware of that boils and / or ulcers, such as diabetic ulcers, pressure ulcers and / or boils may be particularly difficult to heal. Thus, in one embodiment, said ulcer is selected from the group consisting diabetic ulcers, boils and pressure ulcers, such as bed sores. In one embodiment, said ulcer is selected from the group consisting diabetic ulcers and pressure ulcers, such as bed sores. In one particular embodiment, said ulcer is a pressure ulcer. In another embodiment, said wound is a boil, such as an open boil.

[0140] In one embodiment, said wound is an acute wound. Moreover, as discussed above, the present inventors show that compositions according to the disclosure are particularly beneficial in the treatment of chronic wounds. Accordingly, in another embodiment, said wound is a chronic wound. In yet another embodiment, said wound is an infected wound. The skilled person appreciates that infected wounds may be difficult to heal and become chronic. Thus, in yet another embodiment, said wound is a chronic infected wound. In some embodiments, the wound is selected from the group consisting of diabetic ulcers, pressure sores, burn injuries, surgical incisions and infected wounds.

[0141] It is envisioned, that the compositions and / or pharmaceutical formulations as disclosed herein may be administered by one of the routes as explained in relation to the fourth aspect of the disclosure.

[0142] In one embodiment, the discussed uses of the composition or the pharmaceutical formulation comprise topical administration of said composition or said pharmaceutical formulation to a patient in need thereof. Said patient may be for example an animal. Said patient may be a human. In one embodiment, said administration is a one-time administration or a repeated administration.

[0143] Without being bound by theory, it is envisioned that it is beneficial that the composition is in contact with the wound for a significant time, such as at least two hours or more, such that the active agent(s) can exert their effects. In one embodiment, said composition or said pharmaceutical formulation is in contact with said wound for at least approximately 8 hours, such as at least approximately 10 hours, such as at least approximately 12 hours, such as at least approximately 18 hours, such as at least approximately 24 hours, per administration.

[0144] In another embodiment said composition or said pharmaceutical formulation is in contact with said wound between 8 hours to 48 hours, such as between 12 hours to 48 hours, such as between 12 hours to 36 hours, such as between 12 hours to 24 hours, per administration.

[0145] The skilled person will appreciate that the composition according to the disclosure may be in contact with the wound for a less than 8 hours per administration depending on the patient’s need. Moreover, the composition according to the disclosure may be in contact with the wound for longer than 24 hours per administration depending on the patient’s need. In one embodiment, the wound is an infected wound. In one embodiment, visible signs of the infection have disappeared after one administration, such as approximately after 24 hours. In one embodiment, visible signs of the infection have disappeared after two administrations, such as approximately after 48 hours.

[0146] Without bound by theory, it is envisioned that the duration of an administration using a composition according to the disclosure may be shorter in case of children and / or young adults, due to their ability to heal faster than adults and / or elderly people. Furthermore, it is envisioned that the number of administrations and / or the duration of each individual administration event may be chosen based on the type of the wound to be treated and / or adjusted according to the progression of wound healing during a treatment using a composition as disclosed herein. It is furthermore envisioned that that the number of administrations and / or the duration of each individual administration event may be adjusted according to the age and / or medical condition(s) of the patient to be treated using a composition as disclosed herein.

[0147] In certain embodiments, there is a time interval between said repeated administrations, wherein said composition or said pharmaceutical formulation is not administered to said patient.

[0148] Said time interval may be at least 30 min, such as wherein said time interval is approximately 30 min. It is appreciated that the duration of said time interval may be adjusted according to the needs of said patient and / or the type of the wound.

[0149] In some embodiments, during said time interval a disinfection of the wound is performed. Thus, for example, it is appreciated that said time interval may be adjusted to the duration that is needed for said disinfection. In some embodiments, a disinfection of the wound is performed prior to said administration. The skilled person appreciates that this may be particularly useful in case of the treatment of infected wounds. Thus, the treatment may comprise an initial disinfection of the wound and optionally disinfections between at least two consecutive administrations of the composition. In some embodiments, one or more disinfection of the wound is performed after a last administration of said composition or said pharmaceutical formulation. It is appreciated that said disinfection may be beneficial for preventing an occurrence and / or a reoccurrence of an infection.

[0150] Any suitable wound disinfection agent may be used. In certain embodiments, wherein disinfection occurs, the wound is disinfected by using an oil, such as an oil derived from the species Pistacia atlantica, such as wherein the oil is derived from the seeds of the species Pistacia atlantica.

[0151] The skilled person appreciates that a disinfecting agent other than the Pistacia atlantica tree oil may be used for the disinfection of a wound.

[0152] As discussed above a composition or a pharmaceutical formulation as described herein, may be beneficial for use in tissue regeneration.

[0153] Thus, in a related seventh aspect of the present disclosure there is provided a composition or a pharmaceutical formulation as described herein, for use in in tissue regeneration. Said tissue regeneration may be re-epithelialization. As demonstrated in the appended Examples, the composition as disclosed herein promotes tissue regeneration, such as re-epithelialization. It will be appreciated that the embodiments disclosed in connection with the sixth aspect, including embodiments related to dose, frequency of administration and route of administration, are equally relevant for this seventh aspect and are not repeated here merely for the sake of brevity.

[0154] In a related eighth aspect, there is provided a method for treatment of a wound comprising administration of a composition, or a pharmaceutical formulation as described herein, to a patient in need thereof. It will be appreciated that the embodiments disclosed in connection with the sixth aspect, including embodiments related to dose, frequency of administration and route of administration, are equally relevant for this eighth aspect and are not repeated here merely for the sake of brevity.

[0155] In one embodiment of the method, said wound is a penetrating wound, a boil or an ulcer.

[0156] In one embodiment, said wound is a penetrating wound. Said penetrating wound may be selected from the group consisting of a surgical wound and / or an incision, a bum, a bite, an abrasion, a laceration and a skin tear. Said penetrating wound may be a burn.

[0157] In one embodiment, said wound is an ulcer.

[0158] Said ulcer may be selected from the group consisting of peptic ulcers, mouth ulcers, arterial ulcers, venous ulcers, pressure ulcers, diabetic ulcers and genital ulcers.

[0159] Said ulcer may be selected from the group consisting of peptic ulcers and mouth ulcers, such as gastric ulcers and / or duodenal ulcers.

[0160] Said ulcer may be selected from the group consisting diabetic ulcers and pressure ulcers, such as bed sores.

[0161] In one embodiment, said wound is a boil, such as an open boil.

[0162] In certain embodiments, said wound is an acute wound.

[0163] In other embodiments, said wound is a chronic wound.

[0164] Said wound may be an infected wound.

[0165] In one embodiment there is provide the method for treatment as disclosed herein, wherein said administration is topical administration. Said administration may a one-time administration. Moreover, said administration may be a repeated administration.

[0166] In one embodiment there is provide the method for treatment as disclosed herein, wherein said composition or said pharmaceutical formulation is in contact with said wound for at least approximately 8 hours, such as at least approximately 10 hours, such as at least approximately 12 hours, such as at least approximately 18 hours, such as at least approximately 24 hours, per administration.

[0167] In another embodiment, said composition or said pharmaceutical formulation is in contact with said wound between 8 hours to 48 hours, such as between 12 hours to 48 hours, such as between 12 hours to 36 hours, such as between 12 hours to 24 hours, per administration.

[0168] In certain embodiments there is provide the method for treatment as disclosed herein, wherein there is a time interval between said repeated administrations, wherein said composition or said pharmaceutical formulation is not administered to said patient.

[0169] Said time interval is at least 30 min, such as wherein said time interval is approximately 30 min.

[0170] In an embodiment, during said time interval a disinfection of the wound is performed.

[0171] In another embodiment, a disinfection of the wound is performed prior to said administration.

[0172] In yet another embodiment, one or more disinfection of the wound is performed after a last administration of said composition or said pharmaceutical formulation.

[0173] Said wound may be disinfected by using an oil, such as an oil derived from the species Pistacia atlantica, such as wherein the oil is derived from the seeds of the species Pistacia atlantica.

[0174] As discussed above a composition or a pharmaceutical formulation as described herein, is considered by in the inventors to be beneficial for use in tissue regeneration, such as for tissue regeneration by re-epithelialization. Thus, in a related nineth aspect of the present disclosure there is provided a method for tissue regeneration comprising administration of a composition as, or a pharmaceutical formulation as described herein, to a patient in need thereof.

[0175] It will be appreciated that the embodiments disclosed in connection with the sixth aspect, including embodiments related to dose, frequency of administration and route of administration, are equally relevant for this nineth aspect and are not repeated here merely for the sake of brevity.

[0176] In a related tenth aspect of the present disclosure there is provided a use of

[0177] - a component of herb A, wherein said herb A is selected from a species of the genus Pistacia;

[0178] - a component of herb B, wherein said herb B is selected from a species of the genus Salvia; and

[0179] - a biomatrix of animal-origin, in the manufacture of a medicament for the treatment for the treatment of wounds.

[0180] In one embodiment, there is provided a use of

[0181] - an herbal extract derived from a component of herb A, wherein said herb A is selected from a species of the genus Pistacia',

[0182] - an herbal extract derived from a component of herb B, wherein said herb B is selected from a species of the genus Salvia', and

[0183] - a biomatrix of animal-origin.

[0184] In the manufacture of a medicament for the treatment of wounds.

[0185] It will be appreciated that the embodiments disclosed in connection with the sixth aspect, including embodiments related to dose, frequency of administration and route of administration, are equally relevant for this tenth aspect and are not repeated here merely for the sake of brevity.

[0186] It is noted that, as used in this specification and the appended claims, the singular forms “a”, “an”, and “the” also include plural referents unless the context clearly dictates otherwise.

[0187] As used herein, when the term “about” or “approximately” is used in relation to a numerical value, it is to be interpreted as a range of ± 10 %, such as ± 9 %, such as ± 8 %, such as ± 7 %, such as ± 6 %, such as ± 5 %, such as ± 4 50%, such as ± 3 %, such as ± 2 %, such as ± 1 %. For example, when the value is stated to be about 10, this means that the value is in fact in the range of from 9 to 11 , such as in the range of from 9.9 to 10.9, such as in the range of from 9.8 to 10.8, such as in the range of from 9.7 to 10.7, such as in the range of from 9.6 to 10.6, such as in the range of from 9.5 to 10.5, such as in the range of from 9.4 to 10.4, such as in the range of from 9.3 to 10.3, such as in the range of from 9.2 to 10.2, such as in the range of from 9.1 to 10.1.

[0188] The skilled person knows that numerical values relating to measurements are subject to measurement errors which place limits on their accuracy. For this reason, the general convention in the scientific and technical literature is applied: the last decimal place of a numerical value indicates its degree of accuracy. Where no other error margins are given, the maximum margin is ascertained by applying the rounding-off convention to the last decimal place e.g. for a measurement of 3.5 cm, the error margin is 3.45-3.54. When interpreting ranges of values in patent specifications, the skilled person proceeds on the same basis.

[0189] While the invention has been described with reference to various exemplary aspects and embodiments, it will be understood by those skilled in the art that various changes may be made and equivalents may be substituted for elements thereof without departing from the scope of the invention. Therefore, it is intended that the invention is not limited to any particular embodiment contemplated, but that the invention will include all embodiments falling within the scope of the appended claims. The invention will be further illustrated by the following non-limiting Examples.

[0190] BRIEF DESCRIPTION OF THE FIGURES

[0191] Figure 1 shows images of burn wound on the left leg of a 3.5-year-old male patient treated with the inventive composition. Images were taken on day 0 and day 12.

[0192] Figure 2 shows images of bum wound on the left foot of a 3.5-year-old male patient treated with the inventive composition. Images were taken between day 12 and day 70.

[0193] Figure 3 shows images of ulcer, a chronic wound, of a 55-year-old female patient, treated with the inventive composition. Images were taken between day 1 and 5.

[0194] Figure 4 shows representative images from the wound healing process in a representative animal treated with the composition according to the invention over time. Images from Day 0, 1 , 2, 3, 5, 7, 9, 11 and 14 are shown, the untreated control is presented on the left hand side and the treated wound on the right side.

[0195] Figure 5 shows a bar graph of the morphological assessment of the degree of healing of the wounds on Day 5, 10 and 13. Treated and untreated wounds were compared for each of comparative control compositions AC, AB and BC as well as for composition ABC according to the present invention. Score values are shown on the y-axis and the compositions are presented on the x- axis.

[0196] Figure 6A-D shown representative microscopy images of hematoxylin-eosin stained tissue sections obtained at Day 5 and Day 10 as described in Example 8. Images of wounds treated with comparative control compositions AC, AB or BC; with the composition ABC according to the present invention as well as untreated control wounds are presented in the figure. Black triangle indicates the wound egde and dashed line indicates re-epithelialization. Bar graphs of wound re-epithelialization as well as wound contraction at each time point assessed are presented. * means p < 0.05, ** means p < 0.01 , *** means p < 0.001 and ***’ means p < 0.0001 .

[0197] EXAMPLES

[0198] The following Examples illustrate a composition according to the disclosure for use in the treatment of wounds. Example 1 presents a preparation method for a composition, which is used in the treatment of a burn and an ulcer as described in the case studies according to Examples 2 and 3, respectively. Moreover, Example 4 presents an example, wherein said composition according to Example 1 is tested in an animal model for wound healing.

[0199] Example 1

[0200] A preparation method for a composition according to the disclosure A composition according to the disclosure was prepared according to the following procedure: Materials and Methods

[0201] A) Turpentine gum of the species P. atlantica was collected from P. atlantica trees wildly grown in the province Balochistan in Iran. The turpentine gum was placed in a freezer for 24 hours at a freezing temperature of -8 °C, and thus, frozen turpentine gum was obtained. Following this freezing step, the turpentine gum was ground and then sieved using a mesh having a mesh size of less than approximately 1 mm. By this process, a mechanically processed turpentine gum was obtained with a homogenous particle size less than approximately 1 mm. Thus, said processed turpentine gum had a powder-like texture. In other words, said processed turpentine gum is a turpentine gum powder. The turpentine gum was not cleaned using any aqueous solution, such as water, during this process.

[0202] B) Leaves of the species S. mirzayanii were collected, wherein said S. mirzayanii was wildly grown in the province Balochistan in Iran. The leaves were separated from the stem and the thorns of the herb. The leaves were heated in a pan at a heating temperature of approximately 170 to 180 °C until the leaves became completely dry. The duration of this drying step was approximately 1 min. Drying is useful for the removal of the water content of the leaves. The drying step was continued until the color of the leaves turned dark brown.

[0203] C) Omentum, also called as belly lace fat (referred to pie tori shekam boz, “ in Persian), obtained from an animal of the species Capra aegagrus hircus, was briefly rinsed with water and then placed in a freezer for 24 hours at a freezing temperature of -8 °C to obtain frozen omentum.

[0204] The above processed components were mixed thoroughly in a weight ratio of 1 :1 :1 .5, i.e. in a weight ratio corresponding to one portion of turpentine gum (A), one portion of leaves (B) and one and a half portion of omentum (C). The thereby obtained mix was ground to obtain a homogeneous mixture using an electric mixer. The grinding resulted in a mixture with even distribution of each component in the mixture. Then the ground mixture was kept in the refrigerator for approximately 72 hours at a cooling temperature of approximately 4 to 5 °C. Following the cooling step, the mixture was ground one more time to obtain a reprocessed mixture. The duration of this second grinding step was approximately 30 min. The reprocessed mixture was a homogeneous mixture of the components. Said reprocessed mixture was a gray, medium-hard solid paste with molding ability.

[0205] Results and Conclusions

[0206] The composition had a paste like texture. Moreover, it can easily be formed, for example rolled out and / or cut, to have a desired size. The desired size may be selected based on the type and / or the extension of a wound to be treated. The composition can be shaped for example to have a bandage-like shape or a plaster-like shape for topical administration or can be shaped as a pill for oral administration. Said composition is edible.

[0207] Example 2

[0208] Case study I - Treatment of a severe burn

[0209] Example 2 illustrates the successful treatment of a wound using the composition according to the invention as described herein. A composition was prepared according to Example 1 , and used for the treatment of a burn wound as described below.

[0210] Materials and Methods

[0211] Patient:

[0212] A 3.5-year-old male patient, otherwise healthy, suffered from a car accident and injured a severe bum on the left leg. The bum extended on the thigh (from the knee to the hip) of said patient. The bum was a deep and open wound (Day 0, Fig. 1A). The wound was characterized by a severe tissue damage, such as a severe destruction of the tissue in the affected area. Said wound was also infected.

[0213] Treatment (Day 0 to Day 70): The wound was disinfected using P. atlantica tree oil derived from the seeds of said tree. The wound was cleaned and pus from the damaged tissue was removed. Following the disinfection step, a composition as prepared in Example 1 was placed on the wound of the thigh for 24 hours (referred to as administration event 1 ). During this process, the composition was kept in contact with the wound.

[0214] After 24 hours, the used composition was removed from the wound and the wound were disinfected using P. atlantica tree oil derived from the seeds of said tree. The wound was cleaned and pus from the damaged tissue was removed.

[0215] Approximately 30 minutes after removal of the composition administrated during administration event 1 , a second batch of the composition as prepared in Example 1 was placed on the wound of the thigh for 24 hours (referred to as administration event 2). During this process, the composition was kept in contact with the wound.

[0216] After 24 hours, the used composition was removed from the wound and the wound was disinfected using P. atlantica tree oil derived from the seeds of said tree. The wound was cleaned and pus from the damaged tissue was removed.

[0217] Following the above, the wound was treated by the repeated administration of a new batch of the composition as prepared in Example 1 , wherein during each administration the composition was kept in contact with the wounds for approximately 12 hours. There was an approximately 30 min long interval between each administration event, wherein a composition according to the disclosure was not administered to the patient. During this time interval, the wound was examined and disinfection and / or cleaning of the wound as described above was performed if it was deemed necessary.

[0218] The above treatment process using repeated topical administration of a composition according the disclosure was stopped at Day 12 (Fig 1 B), when the depth of the wound reached approximately 0.2 mm in relation to the surrounding undamaged area.

[0219] Following the administration events comprising topical administration of the composition according present invention as described above, the wound was treated with P. atlantica tree oil derived from the seeds of said tree every 24 hours, until the wound was no longer susceptible for infections.

[0220] Results

[0221] The healing of the wound during the above treatment process is depicted in Fig. 1 and 2. The composition reduced and then diminished the infection of the wounds. The composition promoted regeneration of the injured tissues. Extensive tissue regeneration occurred within a short time (Fig 1 A and 1 B). Tissue of the skeletal muscles and skin was regenerated. It was concluded that revascularization of the tissue and regeneration of the nerves also occurred, as physiological functions of the regenerated tissue were corresponding to that of the surrounding undamaged tissue of the limb. The treatment induced no adverse side effects.

[0222] Conclusions

[0223] The present case exemplifies the benefits of a composition according to the invention. Said composition is advantageous in the treatment of wounds, in particular, in the treatment of severe and deep wounds with substantive tissue damage. It is concluded that said composition exhibited antimicrobial activity, such as antibacterial activity. It was also concluded that said composition exhibited anti-inflammatory activity. Moreover, said composition is advantageous in promoting tissue regeneration. Importantly, the composition is also advantageous in promoting tissue regeneration in case of deep wounds with severe tissue damage. The present inventors consider that said composition is useful in wound healing. In particular, the composition is useful in reducing the time needed for the healing of a wound. Without being bound by theory, it is believed that the wound-healing effect of said composition according to the disclosure may manifest in two stages if a wound to be treated is infected. In such cases, the composition may diminish infection of a wound in a first stage. In a second stage, the composition may promote tissue regeneration of the disinfected wound. It is believed that the composition prevents occurrence and / or reoccurrence of an infection of the wound during the treatment process. Such prevention may be achieved by the antibacterial activity, such as antimicrobial activity, of the composition.

[0224] Example 3

[0225] Case study II - Treatment of an ulcer

[0226] Example 3 illustrates the successful treatment of an ulcer using the composition according to the invention as described herein. A composition was prepared according to Example 1 , and used for the treatment of an ulcer, more specifically a pressure ulcer (also known as bedsore).

[0227] Materials and Methods

[0228] Patient:

[0229] A 55-year-old female patient, diagnosed with severe diabetes, developed an ulcer located on the inner thigh of said patient. The patient had been hospitalized and the ulcer had been surgically treated. More than 40 days after the surgery the ulcer was not healed. In other words, said ulcer was a chronic wound.

[0230] Treatment method:

[0231] A composition as prepared in Example 1 was placed on the ulcer for 24 hours (referred to as administration event 1 ). During this process, the composition was kept in contact with the ulcer.

[0232] After 24 hours (Day 1 , Fig. 3A), the used composition was removed from the ulcer. Approximately 30 minutes after the removal of the composition applied during administration event 1 , a second batch of the composition as prepared in Example 1 was placed on the ulcer for 24 hours (referred to as administration event 2), wherein the composition was kept in contact with the ulcer.

[0233] After 24 hours (Day 2, Fig. 3B), the used composition was removed from the ulcer. Approximately 30 minutes after the removal of the composition applied during administration event 2, a third batch of the composition as prepared in Example 1 was placed on the ulcer for 24 hours (referred to as administration event 3), wherein the composition was kept in contact with the ulcer.

[0234] After 24 hours (Day 3, Fig. 3C), the used composition was removed from the ulcer. Approximately 30 minutes after the removal of the composition applied during administration event 3, a fourth batch of the composition as prepared in Example 1 was placed on the ulcer for 24 hours (referred to as administration event 4), wherein the composition was kept in contact with the ulcer.

[0235] After 24 hours (Day 4, Fig. 3D), the used composition was removed from the ulcer. The depth of the ulcer reached approximately 0.1 mm in relation to the surrounding undamaged area and no further administration events occurred. Fig. 3E shows the ulcer 24 hours after the removal of the last administered composition.

[0236] It is to be understood that during the approximately 30 minutes-long intervals between the administrations, a composition according to the disclosure was not administered to the patient. During the approximately 30 minutes-long intervals, the ulcer was inspected but cleaning and / or disinfection of the ulcer was deemed unnecessary.

[0237] Results

[0238] The healing of the ulcer during the above treatment process is depicted in Fig. 3. The composition reduced and then diminished the infection of the ulcer. The composition promoted regeneration of the injured tissues. Wound healing and tissue regeneration occurred within a short time. The treatment induced no adverse side effects.

[0239] Conclusions

[0240] It is appreciated that certain medical conditions, such as diabetes, predispose a patient to a condition, wherein said patient develops chronic wounds, such as ulcers, and / or the healing duration of a wound of said patient may be prolonged substantially due to the existence of said medical condition. It is apparent from the present results that a composition according to the invention is useful in the treatment of chronic wounds. Specifically, said composition is beneficial in the treatment of wounds, wherein the healing duration of a wound is normally prolonged. Moreover, said composition is useful in the treatment of wounds, wherein said wounds do not respond to any other treatment.

[0241] Example 4

[0242] Hypothetical Example - Animal models of wound healing

[0243] There are several preclinical models by which therapeutic effects of a composition as disclosed herein may be measured and confirmed (for example as disclosed in Chen et al. 2013, Methods Mol Biol. 1037:265-274; Masson-Meyers et al. 2020, Int. J. Exp. Path. 101 :21-37 and Grada et al. 2018, J. of Investigative Dermatology 138:2095-2105). Non-limiting examples are experimental animal models, such as rodents, which are predictive for studying human wound healing in vivo.

[0244] In brief, the experiments for creating wound models for evaluating and confirming the therapeutic effects of a composition as disclosed herein are carried out as follows:

[0245] General considerations

[0246] Primarily full-thickness wound models are used, wherein the wounds extend through all layers of the skin including the dermis, epidermis and the subcutaneous tissue as well as the panniculus carnosus. It is to be understood that in case of the following excisional wound models, in some instances assessment of wound healing may occur in full-thickness wound models, wherein the wounds extend into the skeletal muscle tissue to assess healing of severe wounds.

[0247] Preferably, in case of incisional and excisional wound models, wherein the wounds are created on the dorsum of the animals, at least two wounds are created, thus each animal can act as its own internal control, enabling the application of both the treatment and control on the same animal.

[0248] To minimize the effect of hair follicles on wound healing, the wounds are preferably created during the telogen phase of the hair cycle. It is to be understood that adult C57BL / 6 mice used in the following nonlimiting examples may be adapted for a more desired age and / or more desired strain. Moreover, a wild-type mouse model may be exchanged to a desired transgenic mouse model. As such, e.g. young and / or aged mice may be used to assess wound healing which recapitulates wound healing in young and / or elderly human subjects. Furthermore, transgenic animal models which correspond to a pre-existing condition which affects wound healing (such as diabetes) may be used to assess wound healing recapitulating such conditions in human. Non-limiting example for the use of such transgenic models is the below diabetic and obesity models as well as e.g. IL-6 KO mice which are characterized by significantly delayed wound healing.

[0249] A) Incisional model

[0250] Incisional models are considered to resemble acute wounds. Adult mice, e.g. seven- to eight-week-old male C57BL / 6 mice, are anesthetized, and the dorsal flank is shaved and sterilized with an alcohol swab. Two linear 1 cm scalpel full-thickness incision are made 1 cm apart on the dorsum of mice and left to heal by secondary intention, that is the skin edges are not sutured together. Postoperatively, mice are housed individually to minimize wound disruption with access to food and water ad libitum.

[0251] B) Excisional model

[0252] Excisional wound models are the most commonly used wound healing models. They are considered clinically relevant for both acute wounds as well as chronic wounds, such as non-healing ulcers. This model allows e.g. the investigation of hemorrhage, inflammation, granulation tissue formation, re- epithelialization, angiogenesis and re-modelling.

[0253] B1) Non-splinted excision model

[0254] Adult mice, e.g. seven- to eight-week-old male C57BL / 6 mice, are anesthetized, and the dorsal flank is shaved and sterilized with an alcohol swab. Two full-thickness circular 6 mm biopsy punch excisions are made 1 cm apart on the dorsum of mice and left to heal by secondary intention, that is the skin edges are not sutured together. Postoperatively, mice are housed individually to minimize wound disruption with access to food and water ad libitum.

[0255] B2) Splinted excision model

[0256] In this model, a splinting technique is used to minimize myofibroblast- mediated contraction of the wound, and to allow healing through granulation and re-epithelization similar to humans.

[0257] Adult mice, e.g. seven- to eight-week-old male C57BL / 6 mice, are anesthetized, and the dorsal flank is shaved and sterilized with an alcohol swab. Two full-thickness circular 6 mm biopsy punch excisions are made 1 cm apart on the dorsum of mice and left to heal by secondary intention, that is the skin edges are not sutured together. Silicon splints having an outer diameter of 16 mm and an inner diameter of 10 mm are glued and secured concentrically around the wound by evenly spaced interrupted stiches. Postoperatively, mice are housed individually to minimize wound disruption with access to food and water ad libitum.

[0258] C) Mouse tail model

[0259] This model is used to recapitulate delayed wound closure in animals. In comparison to back dorsal wounds, tail wounds require up to 21 days for full resurfacing, thus they are suitable to assess treatment of the wound in an extended timeframe.

[0260] Adult mice, e.g. seven- to eight-week-old male C57BL / 6 mice, are anesthetized, and the tail is shaved and sterilized with an alcohol swab. A rectangular (0.3 x 1 cm) full-thickness excision is made 1 cm distal to the body of the mouse on the dorsal aspect of the tail. Postoperatively, mice are housed individually to minimize wound disruption with access to food and water ad libitum.

[0261] D) Ischemia reperfusion model

[0262] Pressure ulcers are thought to be mediated by cycles of ischemia followed by reperfusion injury. This model is thus clinically relevant for pressure ulcers and the method described herein can achieve a stage III pressure ulcer. In principle, two ceramic magnetic disks are placed to “pinch” the dorsal skin and apply 50 mmHg of pressure which has been shown to decrease blood flow by 80 %. This is followed by removal of the magnets to allow for a period of reperfusion injury.

[0263] Adult mice, e.g. seven- to eight-week-old male C57BL / 6 mice, are anesthetized, and the dorsal flank is shaved and sterilized with an alcohol swab. Two ceramic magnets are placed made 1 cm apart on the dorsum of mice, each 5 mm from the mid-line, so that a fold of skin is pinched between the magnets. The skin fold includes epidermis, dermis, subcutaneous fat, panniculus carnosus, and subcutaneous loose connective tissue layer (hypodermis), but not muscle. After 12 hours the magnets are removed from each mouse. The 24-hour ischemia-reperfusion cycle is repeated for a total of three cycles. During the ischemia-reperfusion cycles and postoperatively, mice are housed individually to minimize wound disruption with access to food and water ad libitum. Ulcers become visible after 3-4 days and reach a maximum depth at 10 days.

[0264] E) Ischemic flap model

[0265] In this model, a three-sided full thickness peninsular flap is created on the dorsum of the animals, wherein an impermeable sheet is placed under the flap to prevent neovascularization from below the wound, leaving only the flap pedicle as a vascular source. This model is clinically relevant to assess neovascularization, which consists of both angiogenesis (sprouting of new vessels from existing endothelium) as well as vasculogenesis (creating of de novo vessels by progenitor cells).

[0266] Adult mice, e.g. seven- to eight-week-old male C57BL / 6 mice, are anesthetized, and the dorsal flank is shaved and sterilized with an alcohol swab. A longitudinal rectangle (10 mm x 25 mm) is marked on the dorsal midline, wherein the cranial edge of the rectangle is at the level of the shoulder blades. Along the caudal and longitudinal edges of the rectangle, the skin is incised and the cranial edge is kept intact as the pedicle. A fullthickness peninsular flap is raised. A 10 mm x 25 mm silicone sheet is placed within the wound between the muscle and the skin, and moist is kept in the wound by periodically wetting with PBS. The flap is sutured with interrupted stiches. Postoperatively, mice are housed individually to minimize wound disruption with access to food and water ad libitum. The flap does not develop necrosis but remains ischemic for up to 2 weeks.

[0267] F) Burn wound model

[0268] Adult mice, e.g. seven- to eight-week-old male C57BL / 6 mice, are anesthetized, and the dorsal flank is shaved and sterilized with an alcohol swab. A burn wound of the dorsal skin is created by direct application of heat to the skin by a hot metal plate which induces damage of all skin layers.

[0269] G) Diabetic wound model

[0270] The above (A-F) procedures are repeated using diabetic mouse models to assess wound healing in models which is clinically relevant for wound healing under a pre-existing condition which delays and / or prevents the healing process.

[0271] For this purpose, transgenic animal models of type 2 diabetes, insulin resistance and / or obesity are used, such as nonobese diabetic (i.e. NOD) mice, leptin receptor-deficient diabetic mice (db / db) and obese / obese (ob / ob) mice.

[0272] Moreover, animal models wherein hyperglycemia and / or type 1 diabetes is chemically induced using intraperitoneal or caudal vein injection of alloxan or streptozotocin, which induce [3-cell necrosis, are also tested. These animals are allowed to manifest hyperglycemia for several weeks prior to creating a wound as described herein. Furthermore, animal models wherein metabolic syndrome and / or obesity is induced by high fat diet are also tested.

[0273] In these cases, the wound creation methods are adapted according to standard experimental observations if needed, for example in case of db / db mice, when an ischemia reperfusion model is applied for wound creation, 12- hour ischemia-reperfusion cycles are applied to induce pressure ulcers.

[0274] H) Infected wound model

[0275] A characteristic of chronic human wounds is bacterial infection of the wounds. Infection of the wounds normally delays and / or inhibits the healing process, which enables assessment of the therapeutic effect of an inventive composition as disclosed herein when applied to wounds, which take long time to heal or tend to recur. Thus, the above (A-G) procedures are repeated and after the described wound creation, a bacterial suspension, such as a bacterial suspension of Pseudomonas aeruginosa or Streptococcus aureus, is applied to the surface of the wound. Bacterial concentration is adjusted according to pathogenicity, virulence, and the extent of the immune response of the host following standard protocols.

[0276] Conclusions

[0277] The above non-limiting examples of wound healing mouse models recapitulate different types of wounds subjected to the present disclosure and are expected to be suitable for evaluating and confirming the therapeutic effects as disclosed herein.

[0278] Example 5

[0279] Hypothetical Example - Study design and treatment methods

[0280] An inventive composition as disclosed herein, for example a composition according to Example 1 , is administered topically in different wound healing models, wherein wounds are created as described in Example 4. The inventive composition is tested together with appropriate controls, such as according to the following study groups:

[0281] Study groups

[0282] - Study group I (control): non-treated controls

[0283] - Study group II: topical administration of a composition as disclosed herein

[0284] - Study group III (comparative example): topical administration of a composition consisting of a component of herb A and a biomatrix of animal origin as disclosed herein - Study group IV (comparative example): topical administration of a composition consisting of a component of herb B and a biomatrix of animal origin as disclosed herein

[0285] - Study group V (comparative example): topical administration of a composition consisting of a biomatrix of animal origin as disclosed herein

[0286] It is to be understood that Study groups I, III, IV and V are control and comparative example groups, collectively referred to as control groups. Wound healing in Study group II, wherein animals are treated with an inventive composition as disclosed herein, is expected to be accelerated and / or improved in comparison the these control groups.

[0287] In certain experimental setups, an inventive composition according example 1 is applied in Study group II, and compositions according to Study group III, IV and V are prepared according to the method disclosed in Example 1 but lacking the respective components as indicated for the individual control groups.

[0288] In certain experimental setups, for example in case of incisional and excisional wound models, wherein the wounds are created on the dorsum of the animals, at least two wounds are created, thus each animal can act as its own internal control, enabling the application of both the treatment and control on the same animal.

[0289] Treatment

[0290] Topical administration of the compositions according to the above study groups is applied on the wounds created by the different wound models. The length of individual administration events as well as the number of repeated administration events are dependent on the type of the wound to be treated, and are determined experimentally. For example in case of acute wound models, it is expected that shorter administration events and / or fewer administration events are sufficient to achieve improved wound healing in comparison to the treatment methods of a chronic, diabetic and / or infected wound. Thus adapted topical administration methods are applied, such as the following non-limiting examples of treatment processes:

[0291] Treatment process A)

[0292] - wound creation

[0293] - first administration event for 24 hours

[0294] - second administration event for 24 hours

[0295] - third administration event for 12 hours

[0296] - fourth administration event for 12 hours Treatment process B)

[0297] - wound creation

[0298] - first administration event for 12 hours

[0299] - second administration event for 12 hours

[0300] - third administration event for 12 hours

[0301] - fourth administration event for 12 hours Treatment process C)

[0302] - wound creation

[0303] - first administration event for 6 hours

[0304] - second administration event for 6 hours

[0305] Treatment process D)

[0306] - wound creation

[0307] - one administration event for 24, 12 or 6 hours

[0308] It is to be understood that during said administration events the compositions according to the study groups are kept in contact with the wounds, and for each repeated administration event a new composition (i.e. not the removed composition) is placed on the wound. It is also to be understood that in case of non-treated controls, no composition is applied to the wounds, thus the natural healing process can be observed.

[0309] In between repeated administration events the wounds are inspected and documented via in vivo evaluation methods as disclosed in Example 6. It is to be understood that during the documentation periods the compositions are not kept in contact with the wounds.

[0310] Before, during and after the treatment process the wounds are inspected and if deemed necessary, cleaning and / or disinfection of the wound is performed. The need and number of administration events are determined an optimized according to the wound healing process based on the in vivo documentation and inspection of the wounds.

[0311] Conclusions

[0312] It is expected that the above study groups and treatment methods enable evaluation and confirmation of the therapeutic effect of the inventive compositions as disclosed herein in the treatment of wounds, in comparison to appropriate controls as discussed above, as well as in comparison to the normal wound healing process.

[0313] Example 6

[0314] Hypothetical Example - Evaluation methods

[0315] Inventive compositions as disclosed herein, for example as defined in Example 1 , are tested according to the study groups and treatment methods as described in Example 5 using wound models as presented in Example 4. The therapeutic effect of the present inventive compositions in the treatment of wounds is evaluated during the healing process in vivo that allows the monitoring of the wound progression over time, as well as using tissue samples collected from the wounds at different time points of the healing process.

[0316] Non-limiting examples of the evaluation methods are described below, wherein analysis is performed between the study groups using e.g. picture and / or samples collected at the same time point during wound healing:

[0317] A) In vivo visual inspection, documentation and image analysis

[0318] The wounds are evaluated by visual inspection and digital imaging using a standardized photographic protocol at least on a daily base and between administration events. Wound healing and re-epithelization are assessed by wound tracing and calculating the wound healing rate, i.e. the rate of change in the wound surface area, wherein the endpoint of successful treatment is the complete permanent wound closure. Wound healing rate (WHR) or ulcer healing rate (UHR) index are calculated based on [(Ai - Af) / Ai], where Ai represents the initial wound area and Af represents the final area / measurement. Said wound areas are quantified in digital processing of the images taken during the treatment process using appropriate software tools, such as Imaged. The results are expressed in %, wherein 100% means complete re-epithelization while 0% means no signs of re-epithelization.

[0319] It is expected that at a time point when results from Study group II correspond to 100% or near 100% WHR / UHR index, all the other control groups show significantly lower values. The largest difference is expected between the non-treated control group and Study group II.

[0320] During image analysis, not only wound dimensions, such as the surface area and volume (wound depth) are assessed but clinical aspects of the wound healing process are also evaluated, such as by colorimetric analysis for monitoring changes in the surface area by tissue type and its respective colors, such as granulation tissue (red), fibrin / sphacelus (yellow) and necrosis (black).

[0321] It is expected that the analyzed clinical aspects of wounds in Study group II correspond to accelerated wound healing process in comparison to the controls, as well as lack any signs of necrosis.

[0322] B) Sample collection

[0323] Based on the inspection and documentation of the wound healing process in vivo, tissue from the wounds is harvested at different timepoints for further analysis. It is to be understood, that tissue may be harvested as tissue biopsies (e.g. punch biopsies) from living animals, with a method wherein the sample is taken at the edge of the wound for comparison of the wounded area to adjacent healthy tissue. For in depth analysis, animals from different study groups are sacrificed at different time points, wherein the entire wound beds are dissected. Tissue harvesting for analysis of acute phases of wound healing for example occurs at day 0, 3, 5, 7 and 14 postoperatively. It is to be understood that the number of animals in each study group is selected to be sufficient for statistical validity. For in-depth analysis of the wounds, the animals are sacrificed and the wound bed is excised by keeping an approximately 2 mm of adjacent normal tissue. This enables direct comparison of damaged and healthy tissue. The wound is dissected into halves, and one half is further divided into quarters. The larger tissue specimen is fixed e.g. using 4% paraformaldehyde for histological analysis. The two smaller specimens are snap-frozen in liquid nitrogen and are processed for RNA and protein isolation. In certain experimental setups, e.g. for flow cytometry, fresh tissue samples are used for immediate analysis without freezing.

[0324] C) Histopathological analysis and immunohistochemistry

[0325] The fixed tissue specimens are subjected to embedding, sectioning and staining. Tissue sections are stained using hematoxylin-eosin staining and / or trichrome staining to visualize and quantify while blood cells (macrophages, mast cells, lymphocytes and neutrophils) for evaluating the inflammatory phase of wound healing, blood vessels to assess angiogenesis, fibroblasts and collagen to evaluate tissue remodeling and the final scar appearance after wound closure. These staining methods are also useful in evaluating muscle regeneration and keratin levels during the wound healing process. It is expected that results of Study group II will correspond to reduced inflammation, improved angiogenesis and / or more prominent tissue remodeling in comparison to the control groups.

[0326] Immunohistochemistry is applied for detecting and quantifying proteins which are relevant molecular determinants of the wound healing process. Suitable markers to monitor healing are tested using IHC such as collagen I, III and IV, IL-1 [3, IL-10, IL-17, TGF-p, VEGF, cytokeratins 10, 14, vimentin, fibronectin, laminin, a-SMA, CD31 and CD34.

[0327] In is expected that molecular markers promoting wound healing are present at higher levels in tissue samples of Study group II in comparison to the control groups. For example, CD31 and / or CD34 staining indicates increased blood vessel density in tissue sections from Study group II, wherein the wounds are treated with an inventive composition as disclosed herein. D) Immunological methods

[0328] Factors which stimulate endogenous wound repair mechanisms (such as signaling molecules, e.g. cytokines and growth factors produced during wound healing) are identified and quantified using standard immunological methods (such as ELISA) from harvested non-fixed tissue specimens. It is expected an elevated levels of factors which stimulate endogenous wound repair mechanisms are characteristics of the wound healing process in Study group II, and are higher than that of the control groups.

[0329] E) Biochemical and biophysical methods

[0330] Hydroxyproline assay from non-fixed tissue samples is performed according to standard protocols for quantifying hydroxyproline content as biomarker for tissue collagen and collagen turnover. It is to be understood that hydroxyproline may also be evaluated from samples collected post-mortem or from tissue biopsies using chromatography, mass spectrometry, colorimetric and / or enzymatic methods after strong acid or alkali hydrolysis of the tissue samples.

[0331] Collagen is the major component of the connective tissue, and it is expected that in samples from Study group II, the level of hydroxyproline content is increased in comparison to the control groups, indicating increased collagen synthesis and enhanced wound healing.

[0332] Myeloperoxidase (MPO) assay from non-fixed tissue samples is performed according to standard protocols for estimating the recruitment and accumulation of neutrophils and thus evaluating the inflammatory phase of wound healing.

[0333] N-acetylglucosaminidase (NAG) assay from non-fixed tissue samples is performed according to standard protocols for estimating the recruitment and accumulation of macrophages, which are present at all phases of wound healing.

[0334] It is expected that these assay results indicate accelerated and improved wound healing in Study group II in comparison to the results of the control groups. Oxidative stress is assessed during wound healing by measuring biomarkers of cellular stress and antioxidant defense mechanisms for example determined by titration against thiobarbituric acid (TBA) and / or by the ferrous oxidation xylenol orange assay (FOX).

[0335] It is expected that the results of Study group II indicate no or reduced oxidative damage in comparison to the control groups.

[0336] Moreover, flow cytometry and macrophage polarization testing are also applied for evaluating cell recruitment in response to treatment in wounds. For example, such evaluation is performed using markers for endothelial progenitor cells (CD133), endothelial cells (CD31 , CD34 and VEGFR-2), macrophages (CD68), T cells (CD3, CD4, CD8) and B cells (CD20).

[0337] It is expected that the results of these experiments indicate improved and / or accelerated wound healing in contrast to the control groups. As such, a switch between M1 and M2 macrophages (a switch between inflammatory macrophages, which after injury infiltrate the wound and responsible for the cleaning thereof from microorganisms, debris and dead cells, and reparatory macrophages, which along with the infiltration of fibroblasts, keratinocytes and endothelial cells restore the tissue, such as dermis, epidermis and vasculature and orchestrate tissue remodeling) occurs earlier during wound healing in Study group II in comparison to wound healing in the control groups.

[0338] Furthermore, standard methods such as Western blot, and real-time PCR are applied to characterize the presence and amount of biomarkers for wound healing, wherein these methods are expected to reveal biomarkers and increased expression levels thereof indicating improved and / or accelerated healing of wounds, which are treated with an inventive composition as disclosed herein.

[0339] It is to be understood that biomechanical and / or biophysical assessment of wound healing may also be performed to further confirm improved and / or accelerated healing of wounds, which are treated with an inventive composition as disclosed herein, such as diffuse near-infrared spectroscopy (DNIRS) to indicate increased oxygen levels in the blood at the wound site.

[0340] Conclusions

[0341] It is expected that overall wound healing in Study group II, wherein wounds are treated with an inventive composition as disclosed herein, in comparison to the control groups is prominently improved and / or accelerated. It is to be understood that documentation in vivo as well as the analyzed tissue samples are compared from the different study groups at corresponding time points of the wound healing process and the results obtained by the above evaluation methods at each time point indicate improved and / or accelerated wound healing in Study group II in comparison to the control groups.

[0342] For example, in case of certain wounds, a normal wound healing process in mice takes up to 5, 7 or 14 days (e.g. certain acute wounds or chronic wounds), while for example tail wounds require up to 21 days for full resurfacing. It is expected that a treatment using an inventive composition as disclosed herein leads to the acceleration of the wound healing process, thus it takes less time than the normally expected time frames above. This difference is expected to be understood from the comparison of different study groups as described in Example 5. As such, it is expected that study group II exhibits the fastest wound healing process. It is also expected study group II exhibits more desirable values of parameters measured according to the above evaluation methods, which indicate for example improved wound healing and / or accelerated wound healing.

[0343] Example 7

[0344] The purpose of this Example was to assess the effect of the composition according to the present invention on the wound healing process using an excisional wound model in female rats. The assessment included clinical examination, assessment of general appearance as well as macroscopic wound monitoring. Materials and Methods

[0345] Sprague Dawley female rats were sourced from Charles River (Germany). All aspects related to animal housing and care were undertaken in accordance with the national and international regulations concerning animal testing. The food and the water were administered ad libitum during the entire experiment period. The animals were kept under standard conditions of temperature 18- 22°C, humidity 45-55% and in light-controlled conditions (12 h / 12 h light and dark cycles). During duration of the study, the animals were housed into individual ventilated cages.

[0346] Female rats, weighing 200-300 g, 12 weeks age were surgically operated to excise one full-thickness skin patch, in the dorsal region. The animals were anesthetized using isoflurane (3.5%). The back of the animals was shaved, and the selected area was disinfected using 70% ethanol and 0.5% chlorhexidine solution. The animals were placed on the lateral side and two circular full-thickness wound (10 mm in diameter) were made on the dorsum cervical region (one circle on each side of the sagittal axis), using a sterile biopsy skin punch (Kai, Skandivet, Sweden), straight surgical scissors, a tissue forceps and a scalpel blade (no. 24). Two wounds, one of the left side and one of the right side, were made per animal. Bleeding was controlled with gauze compresses until hemostasis.

[0347] The composition according to the present invention was prepared according to Example 1 and is referred to in Table 1 as composition ABC. Comparative control compositions (AB, AC and BC) comprising only two components as specified in Table 1 were prepared analogously. It was noted that composition AB had a different consistency and was less malleable than compositions AC and BC.

[0348] Table 1. Summary of compositions used in study.

[0349] The animals were divided into four experimental groups as outlined below:

[0350] Group 1 : 15 rats - treatment with ABC Group 2: 7 rats - treatment with AB Group 3: 7 rats - treatment with AC Group 4: 7 rats - treatment with BC

[0351] The inventors have attempted to test potential wound healing properties of a composition comprising component C alone in the herein described woundhealing model, but decided for discontinuing the treatment due to ethical reasons and unexpectedly adverse effects on animal health.

[0352] In the experimental set up, the composition (ABC or comparative control AB, AC or BC) was applied to the left wound and the C4 control composition was applied to the right wound of each test animal.

[0353] Wound healing was observed over time and clinical examination of the animals was performed and wounds were measured using a vernier caliper (length and width). Wounds measurements, macroscopic description of the lesions and evaluation of the healing process were performed every 24-48 h over a period of 14 days. To ensure reproducibility, the measurement of the wound area was performed by a single observer throughout the experimental period. Animals were kept until the wounds were healed and were thereafter sacrificed by exposure to carbon dioxide followed by decapitation.

[0354] Results and conclusions All the animals showed good general health condition throughout the study, as assessed by their weight gain (Table 2), food consumption and mobility.

[0355] Table 2. Weight of 10 representative animals in grams over the period of the experiment. Animals were treated with composition ABC.

[0356] Temporarily, some of the animals presented pruritus on the dorsal region, but without interfering with the wound healing process. No signs of intercurrences in wound healing, such as edema, erythema or suppuration in the wound area were observed during the experiment. No signs of infection or inflammation were observed during the whole treatment period.

[0357] Macroscopic images of the wound healing process from a representative animal treated with the composition ABC according to the invention over time are presented in Figure 4. Images from Day 0, 1 , 2, 3, 5, 7, 9, 11 and 14 are shown, the untreated control is presented on the left hand side and the treated wound on the right side.

[0358] It was noted that the wounds treated with the inventive composition or the comparative control compositions exhibited a “build up” of a brownish layer on Day 2. The layer was not characterized as infection and no sign of erythema around the wound was observed.

[0359] Importantly, it was observed that the wounds treated with the inventive composition retained their shape and the tissues covering the site of the wound were generated (as assessed by morphological examination). In contrast, the untreated wounds exhibited a shrinking in wound diameter, which is a commonly observed effect in relation to wound healing in rodents.

[0360] Example 8

[0361] This Example aims to more depth evaluate the wound healing process using an experimental animal model, comparing untreated wounds to those treated with the inventive composition ABC or treated with the comparative compositions AB, AC and BC comparative control (as described in Example 7).

[0362] Materials and Methods

[0363] Sprague Dawley rats (8-10 weeks old) were purchased from a commercial supplier. Rats were housed under standard conditions as follows temperature 18-22°C, humidity 45-55% and in light-controlled conditions (12 h / 12 h light and dark cycles). All aspects related to animal housing and care were undertaken in accordance with the national and international regulations concerning animal testing. Animals were divided to different treatment groups according to Table 3.

[0364] Table 3. Summary of treatment groups.

[0365] The upper dorsal part of each rat was shaved with clippers and sanitized using 70% ethanol. Two full-thickness excisional wounds (approximately 10 mm in diameter) on the dorsal midline of each rat were created. One wound was left untreated as a control (the right side), while the other would was treated with the inventive composition ABC or with one of the comparative compositions (the left side) for three consecutive days after surgery (DO, D1 , D2). The compositions were prepared as described in Example 7 with reference to Example 1 . The animals received analgetic treatment (subcutaneous buprenorfin) for two days and the wound was protected with short stretch bandages (Bastos Viegas s.a.) and large scanpor tape (Norgeplaster). Bandages were avoided when no treatment composition was applied to the wounds to minimize the risk of the bandage sticking to the wound and affecting the healing process.

[0366] In a subset of animals, wound healing was monitored daily by measuring wound size with a caliper and image documentation. The degree of healing of the wounds was evaluated morphologically according to the following categories and score values on the cicatrization below were assigned to each wound:

[0367] Score value 0 No Recovery - Absence of visible tissue regeneration or re- epithel ial ization. The wound remains unhealed with no evidence of dermal or epidermal restoration. Score value 1: Partial Re-epithelialization - Initial signs of epidermal regeneration with partial coverage of the wound bed by new keratinocyte layers, though the skin barrier remains incomplete.

[0368] Score value 2: Advancing Re-epithelialization or Scab Formation - Noticeable formation of a new epithelial layer covering the wound, or presence of a healing eschar (scab), indicating ongoing tissue repair and remodeling.

[0369] Score value 3 Complete Healing or Residual Scab - Full closure of the wound with restored epithelial integrity, minimal to no visible disruption of the skin surface, or presence of a minor residual scab undergoing final desquamation.

[0370] For histological analysis animals were sacrificed on Day 5 or Day 10 postsurgery. The treated and the non-treated wounds together with surrounding tissue were removed carefully with a scalpel and sharp scissors. The wound and the surrounding tissue were fixed in paraformaldehyde (4% PFA) overnight and thereafter transferred to 70% ethanol until further processing. The tissue was embedded in paraffin blocks and sectioned using a microtome at 3-5 pm thickness and mounted on glass slides. Staining using hematoxylin- eosin (H&E) was performed according to standard procedure. Image analysis was performed used Auxioscan 7 (Zeiss).

[0371] The wound contraction and wound re-epithelialization were assessed based on measurement of area. Obtained values were normalized against the control value and are shown in the bar graphs in Figures 6A-D, where the value on the y-axis represents the wound contraction or wound re- epithelialization.

[0372] Results and conclusions

[0373] The degree of healing of the wounds was evaluated morphologically and each sample was assigned a score value between 0 and 3. The results showing the score value assigned the wounds treated with one of the inventive composition ABC, the comparative compositions AB, AC and BC and untreated wounds at Day 5, Day 10 and Day 13 are presented in Figure 5. The data shows that treatment with composition ABC increased the rate of wound healing significantly. At Day 5, wounds treated with composition ABC exhibited a score value of above 2, while wounds treated with either of compositions AB, AC or BC and untreated wounds exhibited a score value of approximately 1 . At Day 10, the ABC treated wounds had reached a value of nearly 3 and at Day 13 the ABC treated wounds were assigned the score value of 3, representing complete healing or residual scap. Taken together, the data shows that wounds treated with the ABC composition exhibited a markedly faster rate of healing compared to wounds treated with the comparative control compositions AB, AC or BC as well as untreaded controls.

[0374] Histological analysis of hematoxylin-eosin stained tissue sections obtained from animals on Day 5 and Day 10 for each of the treatment groups AB, BC, AC and ABC was performed and compared with the untreated controls. Wound re-epithelization as well as wound contraction were assessed.

[0375] As shown in Figure 7A, treatment with the AB composition resulted in significantly improved wound re-epithelization as seen on Day 5 and Day 10. The level of re-epithelization compared to the control was between 1 .2-1 .3 times higher at both evaluated time points. On Day 5 and 10 it was also seen that the treatment with composition AB increased wound contraction and compared to the controls. The presence of eschar was observed.

[0376] As shown in Figure 7B, treatment with the AC composition resulted in significantly higher wound re-epithelization as well as wound contraction compared to the untreated controls. On D5 the wound re-epithelialization of the AC treated wounds was approximately 1 .3 times higher compared to the control, while on Day 10 the level had increased to approximately 1 .6 times. Wound contraction was approximately 1 .4 times higher after AC treatment at both Day 5 and Day 10 compared to the controls. The presence of eschar was observed.

[0377] As shown in Figure 7C, treatment with the BC composition resulted in higher level of wound re-epithelization as well as wound contraction compared to the untreated controls as assessed on Day 5. On Day 10 it was observed that the wound re-epithelization was slightly lower in the BC treated wounds (approximately 1 .2) compared to the controls. On Day 5 and Day 10 it was also seen that the treatment with composition BC increased wound contraction compared to the controls. The presence of eschar was observed.

[0378] As shown in Figure 7D, treatment with the ABC composition according to the invention resulted on Day 5 in significantly improved wound re-epithelization compared to the untreated controls. It is noted that would re-epithelization relative to the control was improved already at Day 5 for the ABC composition (approximately 1.6) compared to an approximate value of 1 .2-1 .3 for the comparative control compositions. At Day 10 wound epithelization was significantly improved (approximately 2.1 times) in comparison to the untreated control and was much higher than that observed for any of the comparative controls. Observed wound contraction was higher in the ABC treated wounds compared to the control and was approximately 1 .4-1 .5 times that of the untreated controls at Day 5 and at Day 10. Importantly, the wound treated with the composition according to the present invention was fully healed already by Day 10. In line with this, an absence of eschar issue was noted.

[0379] Thus, at Day 10 re-epithelialization of wounds treated with comparative control compositions AB, AC or BC was improved in comparison to untreated controls. The wounds treated with compositions AB, AC or BC showed 1 .2- 1 ,5 times more re-epithelialization of the wound compared to the untreated control wounds. Surprisingly, the wounds treated with composition ABC according to the present invention showed strong improved level of re- epithelialized corresponding to more than 2 times of the untreated control wounds. The present inventors conclude that, based on the results herein, treatment with the composition of the present invention significantly improves wound healing and improved re-epithelialization of wounds and results in significantly increased rate of re-epithelialization.

[0380] Example 9

[0381] In this Example, the present inventors assessed inflammatory cell infiltration of wounds treated with the compositions according to the present invention in comparison with untreated control as well as wounds treated with the comparative control compositions.

[0382] Materials and Methods

[0383] Hematoxylin-eosin stained slides of samples obtained as described in Example 8 were digitally scanned and analyzed for the presence of inflammatory cells (particularly neutrophils and macrophages) using the image Processing Platform OpenCV, which employs HSV filtering and morphological segmentation. The digital histological slides were processed using calibrated color thresholds to identify hematoxylin-positive nuclei corresponding to inflammatory cells. Morphological filters enhanced the segmentation of individual cell contours, enabling semi-quantitative cell enumeration. Data from representative images is presented in Table 4a and 4b.

[0384] Results and conclusions

[0385] The analysis was based on semi-automated digital image processing techniques applied to standard H&E-stained slides. While hematoxylin- stained nuclei provide a reasonable approximation of inflammatory cells (particularly neutrophils and macrophages), the method does not differentiate between specific immune cell subtypes with immunohistochemical precision. As such, the reported cell counts are considered a semi-quantitative measurement of relative inflammation. Q7

[0386] As shown in Tables 4a and 4b, the ABC treated samples demonstrated the highest reduction in inflammatory cell count at Day 5 and Day 10, respectively. The data shows a 60% decrease in the number of inflammatory cells in the ABC treated sample relative to the untreated control. It is noted that the inflammatory cell count in samples from untreated controls are in the same range.

[0387] The present inventors conclude that treatment with the inventive composition ABC is the more effective in attenuating inflammation during early wound healing compared to the control compositions as well as to untreated wound. Table 4a. Assessment of number of inflammatory cells at Day 5 in animals 1- 4. Data from treated and untreated wounds are presented for each animal.

[0388] Table 4b. Assessment of number of inflammatory cells at Day 10 for animals 5, 6, 7 and at Day 13 for animal 8. Data from treated and untreated wounds are presented for each animal.

[0389] ITEMIZED LIST OF EMBODIMENTS

[0390] 1 . A composition comprising

[0391] - a component of herb A, wherein said herb A is selected from a species of the genus Pistacia',

[0392] - a component of herb B, wherein said herb B is selected from a species of the genus Salvia', and

[0393] - a biomatrix of animal-origin.

[0394] 2. A composition comprising

[0395] - an herbal extract derived from a component of herb A, wherein said herb A is selected from a species of the genus Pistacia',

[0396] - an herbal extract derived from a component of herb B, wherein said herb B is selected from a species of the genus Salvia', and

[0397] - a biomatrix of animal-origin.

[0398] 3. The composition according to item 1 or 2, wherein said herb A is of the species Pistacia atlantica.

[0399] 4. The composition according to any one of previous items, wherein said herb A is a subspecies of Pistacia atlantica selected from the group consisting of Pistacia atlantica subsp. atlantica, Pistacia atlantica subsp. kurdica, Pistacia atlantica subsp. mutica and Pistacia atlantica subsp. cabulica, such as wherein said herb is of the subspecies of Pistacia atlantica subsp. kurdica. 5. The composition according to any one of the previous items, wherein said herb A is a wild herb or a cultivated herb, such as wherein said herb is a wild herb.

[0400] 6. The composition according to any one of the previous items, wherein said component of herb A is selected from the group consisting of

[0401] - an element of the subterranean subsystem of said herb A;

[0402] - an element of the aerial subsystem of said herb A;

[0403] - an exudate of said herb A; and any combinations thereof.

[0404] 7. The composition according to any one of the previous items, wherein said component of herb A is selected from the group consisting of

[0405] - an element of the aerial subsystem of said herb A;

[0406] - an exudate of said herb A; and any combinations thereof.

[0407] 8. The composition according to any one of the previous items, wherein said component of herb A is an exudate of said herb A.

[0408] 9. The composition according to item 6, wherein said element of the subterranean subsystem is a root, such as a root selected from the group consisting of a taproot, a lateral root, root hairs and any combinations thereof.

[0409] 10. The composition according to item 6 or 7, wherein said element of the aerial subsystem is selected from the group consisting of a stem, a leaf, a flower, a bud, an internode, a node, a fruit, a seed, a trunk, a branch, a twig and any combinations thereof.

[0410] 11 . The composition according to item 10, wherein said element of the aerial subsystem is selected from the group consisting of a leaf, a flower, a fruit and any combinations thereof. 12. The composition according to item 10 or 11 , wherein said element of the aerial subsystem is a fruit.

[0411] 13. The composition according to any one of items 6 to 8, wherein said exudate is an exudate from an element of the subterranean subsystem, which element is a root, such as a root selected from the group consisting of a taproot, a lateral root, root hairs and any combinations thereof.

[0412] 14. The composition according to any one of items 6 to 8, wherein said exudate is an exudate from an element of the aerial subsystem, which element is selected from the group consisting of a stem, a leaf, a flower, a bud, an internode, a node, a fruit, a seed, a trunk, a branch, a twig and any combinations thereof.

[0413] 15. The composition according to item 15, wherein said exudate is an exudate from an element of the aerial subsystem, which element is selected from the group consisting of a trunk, a branch, a twig and any combinations thereof; such as the group consisting of a trunk, a branch and any combinations thereof; such as wherein the element is a trunk.

[0414] 16. The composition according to any one of items 6 to 8 and 13 to 15, wherein said exudate is a resin.

[0415] 17. The composition according to any one of items 6 to 8 and 13 to 15, wherein said exudate is turpentine gum.

[0416] 18. The composition according to any one of the previous items, wherein said component of herb A is processed, such as mechanically processed.

[0417] 19. The composition according to item 18, wherein said component of herb A is processed by freezing and / or grinding. 20. The composition according to any one of the previous items, wherein said herb B is selected from the group consisting of the species Salvia mirzayanii, Salvia macrosiphon, Salvia officinalis and Salvia sharifii, such as the group consisting of the species Salvia mirzayanii, Salvia macrosiphon and Salvia sharifii.

[0418] 21 . The composition according to any one of the previous items, wherein said herb B is of the species Salvia mirzayanii.

[0419] 22. The composition according to any one of the previous items, wherein said herb B is a subspecies of Salvia mirzayanii.

[0420] 23. The composition according to any one of the previous items, wherein said herb B is a wild herb or a cultivated herb, such as wherein said herb is a wild herb.

[0421] 24. The composition according to any one of the previous items, wherein said component of herb B is selected from the group consisting of

[0422] - an element of the subterranean subsystem of said herb B;

[0423] - an element of the aerial subsystem of said herb B; and any combinations thereof.

[0424] 25. The composition according to any one of the previous items, wherein said component of herb B is an element of the aerial subsystem of said herb B.

[0425] 26. The composition according to item 24, wherein said element of the subterranean subsystem is a root, such as a root selected from the group consisting of a taproot, a lateral root, root hairs and any combinations thereof.

[0426] 27. The composition according to item 24 or 25, wherein said element of the aerial subsystem is selected from the group consisting of a stem, a leaf, a flower, a bud, an internode, a node, a fruit, a seed, a trunk, a branch, a twig and any combinations thereof. 28. The composition according to item 27, wherein said element of the aerial subsystem is a leaf.

[0427] 29. The composition according to any one of the previous items, wherein said component of herb B is processed, such as mechanically processed.

[0428] 30. The composition according to item 29, wherein said component of herb B is processed by drying, heating and / or grinding.

[0429] 31 . The composition according to any one of the previous items, wherein said herb B is in a developmental stage selected from the group consisting of sprouting stage, seeding stage, vegetative stage, budding stage, flowering stage and ripening stage; such as the group consisting of flowering stage and ripening stage; such as wherein said herb B is in a ripening stage.

[0430] 32. The composition according to any one of the previous items, wherein said component of herb A exhibits antimicrobial activity and / or anti-inflammatory activity.

[0431] 33. The composition according to item 32, wherein said antimicrobial activity is antibacterial activity.

[0432] 34. The composition according to any one of the previous items, wherein said component of herb B exhibits antimicrobial activity and / or anti-inflammatory activity.

[0433] 35. The composition according to item 34, wherein said antimicrobial activity is antibacterial activity.

[0434] 36. The composition according to any one of the previous items, wherein said biomatrix of animal-origin is a naturally occurring biological material. 37. The composition according to any one of items 1 to 35, wherein said biomatrix of animal-origin is an artificially produced biological material.

[0435] 38. The composition according to any one of the previous items, wherein said biomatrix of animal-origin is an omentum and / or an equivalent thereof, such as wherein said biomatrix of animal-origin is an omentum.

[0436] 39. The composition according to any one of the previous items, wherein said omentum consists of the greater omentum, the lesser omentum or both.

[0437] 40. The composition according to any one the previous items, wherein said biomatrix of animal-origin comprises at least one component of an omentum.

[0438] 41. The composition according to any one of the previous items, wherein said biomatrix of animal-origin is derived from a mammal.

[0439] 42. The composition according to item 41 , wherein said biomatrix of animalorigin is derived from an animal of the family Bovidae, such as of the subfamily Caprinae, such as of the tribe Caprini.

[0440] 43. The composition according to item 42, wherein said biomatrix of animalorigin is derived from an animal of the genus Ovis.

[0441] 44. The composition according to item 43, wherein said biomatrix of animalorigin is derived from an animal of the species Ovis aries.

[0442] 45. The composition according to item 42, wherein said biomatrix of animalorigin is derived from an animal of the genus Capra.

[0443] 46. The composition according to item 45, wherein said biomatrix of animalorigin is derived from an animal of the species Capra aegagrus hircus. 47. The composition according to item 41 , wherein said biomatrix of animalorigin is derived from an animal of the family Rodentia, such as wherein said biomatrix of animal-origin is derived from an animal of the species Mus musculus and / or Rattus norvegicus.

[0444] 48. The composition according to item 41 , wherein said biomatrix of animalorigin is derived from an animal of the order Primates.

[0445] 49. The composition according to item 48, wherein said biomatrix of animalorigin is derived from a human.

[0446] 50. The composition according to any one of the previous items, wherein said biomatrix of animal-origin has tissue regenerative properties, antimicrobial activity and / or anti-inflammatory activity.

[0447] 51 . The composition according to any one of the previous items, wherein said biomatrix of animal-origin has tissue regenerative properties.

[0448] 52. The composition according to any one of the previous items, wherein said biomatrix of animal-origin has antimicrobial activity, such as antibacterial activity.

[0449] 53. The composition according to any one of the previous items, wherein said biomatrix of animal-origin has anti-inflammatory activity.

[0450] 54. The composition according to any one of the previous items, wherein said biomatrix of animal-origin is processed, such mechanically processed.

[0451] 55. The composition according to any one of the previous items, wherein said biomatrix of animal-origin is processed by freezing and / or grinding.

[0452] 56. A preparation method for a composition as defined in any one of the previous items, wherein said method comprises a step of (i) mixing of a component of herb A or an herbal extract thereof as defined in any one of items 1 to 19, 32 and 33; a component of herb B or of an herbal extract thereof as defined in any one of items 1 , 2, 20 to 31 , 34 and 35; and a biomatrix of animal-origin as defined in any one of items 36 to 55; wherein a mixture is obtained.

[0453] 57. The preparation method according to item 56, wherein said method further comprises a step of

[0454] (ii) grinding of the mixture obtained in step (i); wherein a ground mixture is obtained.

[0455] 58. The preparation method according to item 56 or 57, wherein said method further comprises a step of

[0456] (iii) cooling of said mixture obtained in step (i); or of said ground mixture obtained in step (ii).

[0457] 59. The preparation method according to any one of items 56 to 58, wherein said method further comprises a step of

[0458] (iv) grinding of said mixture obtained in step (i); or of said ground mixture obtained in step (ii); or of the cooled mixture obtained in step (iii); wherein a reprocessed mixture is obtained.

[0459] 60. The preparation method according to any one of items 56 to 59, wherein prior to step (i), said method comprises at least one step selected from the group consisting of

[0460] - freezing said component of herb A;

[0461] - grinding of said component of herb A; and

[0462] - sieving of said component of herb A; such as wherein prior to step (i) said method comprises the steps of

[0463] - freezing said component of herb A;

[0464] - grinding of the frozen component of herb A; and

[0465] - sieving of the ground component of herb A 61 . The preparation method according to item 60, wherein the freezing temperature is between approximately -5 to -25 °C, such as between approximately -5 to -15 °C, such as between approximately -5 to -10 °C, such as approximately -8 °C.

[0466] 62. The preparation method according to item 60 or 61 , wherein the freezing duration is at least approximately 1 hour to 24 hours, such as at least approximately 12 hours to 36 hours, such as approximately 24 hours.

[0467] 63. The preparation method according to any one of items 60 to 62, wherein said sieving is performed with a mesh size of approximately 1 mm or less, such as approximately 500 pm or less, such as approximately 250 pm or less, such as approximately 200 pm or less, such as approximately 150 pm or less, such as approximately 100 pm or less, such as approximately 50 pm or less.

[0468] 64. The preparation method according to any one of items 60 to 63, wherein said method does not comprise a cleaning step of said component of herb A, such as a cleaning step using a solution, such as a cleaning step using an aqueous solution, such as water.

[0469] 65. The preparation method according to any one of items 56 to 64, wherein prior to step (i), said method comprises

[0470] - drying said component of herb B.

[0471] 66. The preparation method according to item 65, wherein said drying is performed by heating.

[0472] 67. The preparation method according to item 66, wherein the heating temperature is between approximately 50 °C to 250 °C, such as between approximately 100 °C to 200 °C, such as between approximately 150 °C to 200 °C, such as between approximately 170 °C to 180 °C, such as approximately 175 °C. 68. The preparation method according to any one of items 56 to 67, wherein prior to step (i), said method comprises

[0473] - freezing said biomatrix of animal-origin.

[0474] 69. The preparation method according to item 68, wherein the freezing temperature is between approximately -5 to -25 °C, such as between approximately -5 to -15 °C, such as between approximately -5 to -10 °C, such as approximately -8 °C.

[0475] 70. The preparation method according to item 68 or 69, wherein the freezing duration is at least approximately 1 hour to 24 hours, such as at least approximately 12 hours to 36 hours, such as approximately 24 hours.

[0476] 71 . The preparation method according to any one of items 56 to 70, wherein said grinding in step (ii) and / or (iv) is performed using a mixer with a rotation of at least approximately 5000 RPM, such as between approximately 5000 RPM to 25000 RPM, such as between approximately 10000 RPM to 25000 RPM, such as between approximately 15000 RPM to 20000 RPM.

[0477] 72. The preparation method according to any one of items 56 to 71 , wherein said ground mixture obtained in step (ii) and / or said reprocessed mixture obtained in step (iv) is a homogeneous mixture.

[0478] 73. The preparation method according to any one of items 56 to 72, wherein the cooling temperature in step (iii) is between approximately 0 to 10 °C, such as between approximately 0 to 8 °C, such as between approximately 4 to 8 °C, such as between approximately 4 to 5 °C or such as between approximately 6 to 8 °C.

[0479] 74. The preparation method according to any one of items 56 to 73, wherein the cooling duration in step (iii) is between approximately 1 hour to 96 hours, such as between approximately 24 hours to 96 hours, such as between approximately 48 hours to 96 hours, such as between approximately 60 hours to 80 hours, such as approximately 72 hours.

[0480] 75. The preparation method according to any one of items 56 to 74, wherein the grinding duration in step (ii) and / or (iv) is at least approximately 10 min, such as at least approximately 15 min, such as at least approximately 20 min, such as at least approximately 30 min.

[0481] 76. The preparation method according to any one of items 56 to 75, wherein the grinding duration in step (ii) and / or (iv) is between approximately 10 min to 60 min, such as between approximately 20 min to 40 min, such as approximately 30 min.

[0482] 77. The preparation method according to any one of items 56 to 76, wherein the weight ratio of said component of herb A, said component of herb B and said biomatrix of animal-origin is approximately 1 :1 :1 .5 in step (i).

[0483] 78. A composition according to any one of items 1 to 55, prepared by a process according to any one of items 55 to 77.

[0484] 79. A pharmaceutical formulation comprising a composition as defined in any one of items 1 to 55 and 78 comprising at least one excipient(s).

[0485] 80. The pharmaceutical formulation according to item 79 formulated for topical administration.

[0486] 81 . The pharmaceutical formulation according to item 79 formulated for oral administration.

[0487] 82. A composition as defined in any one of items 1 to 55 and 78, or a pharmaceutical formulation as defined in any one of items 79 to 81 for use as a medicament. 83. A composition as defined in any one of items 1 to 55 and 78, or a pharmaceutical formulation as defined in any one of items 79 to 81 for use in the treatment of a wound, such as for use in wound healing.

[0488] 84. The composition or the pharmaceutical formulation for use according to item 83, wherein said wound is a penetrating wound, a boil or an ulcer.

[0489] 85. The composition or the pharmaceutical formulation for use according to item 83 or 84, wherein said wound is a penetrating wound.

[0490] 86. The composition or the pharmaceutical formulation for use according to item 84 or 85, wherein said penetrating wound is selected from the group consisting of a surgical wound and / or an incision, a bum, a bite, an abrasion, a laceration and a skin tear.

[0491] 87. The composition or the pharmaceutical formulation for use according to any one of items 83 and 86, wherein said penetrating wound is a bum.

[0492] 88. The composition or the pharmaceutical formulation for use according to 83 or 84, wherein said wound is an ulcer.

[0493] 89. The composition or the pharmaceutical formulation for use according to any one of items 83, 84 and 88, wherein said ulcer is selected from the group consisting of a peptic ulcer, a mouth ulcer, an arterial ulcer, a venous ulcer, a pressure ulcer, a diabetic ulcer and a genital ulcer.

[0494] 90. The composition or the pharmaceutical formulation for use according to item 89, wherein said ulcer is selected from the group consisting of a peptic ulcer and a mouth ulcer, such as the group consisting of a gastric ulcer and a duodenal ulcer, such wherein said ulcer is a gastric ulcer. 91 . The composition or the pharmaceutical formulation for use according to item 89, wherein said ulcer is selected from the group consisting of a diabetic ulcer and a pressure ulcer, such as wherein said ulcer is a bed sore.

[0495] 92. The composition or the pharmaceutical formulation for use according to 83 or 84, wherein said wound is a boil, such as an open boil.

[0496] 93. The composition or the pharmaceutical formulation for use according to any one of items 83 to 92, wherein said wound is an acute wound.

[0497] 94. The composition or the pharmaceutical formulation for use according to any one of items 83 to 92, wherein said wound is a chronic wound.

[0498] 95. The composition or the pharmaceutical formulation for use according to any one of items 83 to 94, wherein said wound is an infected wound.

[0499] 96. A composition as defined in any one of items 1 to 55 and 78, or a pharmaceutical formulation as defined in any one of items 79 to 81 for use in tissue regeneration.

[0500] 97. The composition or the pharmaceutical formulation for use according to any one of items 83 to 96, wherein said use comprises topical administration of said composition or said pharmaceutical formulation to the wound of a patient in need thereof.

[0501] 98. The composition or the pharmaceutical formulation for use according to item 97, wherein said administration is a one-time administration.

[0502] 99. The composition or the pharmaceutical formulation for use according to item 97, wherein said administration is a repeated administration.

[0503] 100. The composition or the pharmaceutical formulation for use according to any one of items 97 to 99, wherein said composition or said pharmaceutical formulation is in contact with said wound for at least approximately 8 hours, such as at least approximately 10 hours, such as at least approximately 12 hours, such as at least approximately 18 hours, such as at least approximately 24 hours, per administration.

[0504] 101 . The composition or the pharmaceutical formulation for use according to any one of items 97 to 100, wherein said composition or said pharmaceutical formulation is in contact with said wound between 8 hours to 48 hours, such as between 12 hours to 48 hours, such as between 12 hours to 36 hours, such as between 12 hours to 24 hours, per administration.

[0505] 102. The composition or the pharmaceutical formulation for use according to any one of items 99 to 101 , wherein there is a time interval between said repeated administrations, wherein said composition or said pharmaceutical formulation is not administered to said patient during said time interval.

[0506] 103. The composition or the pharmaceutical formulation for use according to item 102, wherein said time interval is at least 30 min, such as wherein said time interval is approximately 30 min.

[0507] 104. The composition or the pharmaceutical formulation for use according to item 102 or 103, wherein during said time interval a disinfection of the wound is performed.

[0508] 105. The composition or the pharmaceutical formulation for use according to any one of items 97 to 104, wherein a disinfection of the wound is performed prior to said administration.

[0509] 106. The composition or the pharmaceutical formulation for use according to any one of items 97 to 105, wherein one or more disinfection of the wound is performed after a last administration of said composition or said pharmaceutical formulation. 107. The composition or the pharmaceutical formulation for use according to any one of items 104 to 106, wherein said wound is disinfected by using an oil, such as an oil derived from the species Pistacia atlantica, such as wherein the oil is derived from the seeds of the species Pistacia atlantica.

[0510] 108. A method for treatment of a wound comprising administration of a composition as defined in any one of items 1 to 55 and 78, or a pharmaceutical formulation as defined in any one of items 79 to 81 , to a patient in need thereof.

[0511] 109. The method for treatment according to item 108, wherein said wound is a penetrating wound, a boil or an ulcer.

[0512] 110. The method for treatment according to item 108 or 109, wherein said wound is a penetrating wound.

[0513] 111. The method for treatment according to item 109 or 110, wherein said penetrating wound is selected from the group consisting of a surgical wound and / or an incision, a bum, a bite, an abrasion, a laceration and a skin tear.

[0514] 112. The method for treatment according to item 110 or 111 , wherein said penetrating wound is a bum.

[0515] 113. The method for treatment according to item 108 or 109, wherein said wound is an ulcer.

[0516] 114. The method for treatment according to any one of items 108, 109 and 113, wherein said ulcer is selected from the group consisting of a peptic ulcer, a mouth ulcer, an arterial ulcer, a venous ulcer, a pressure ulcer, a diabetic ulcer and a genital ulcer.

[0517] 115. The method for treatment according to item 114, wherein said ulcer is selected from the group consisting of a peptic ulcer and a mouth ulcer, such as the group consisting of a gastric ulcer and a duodenal ulcers, such as wherein said ulcer is a gastric ulcer.

[0518] 116. The method for treatment according to item 115, wherein said ulcer is selected from the group consisting of a diabetic ulcer and a pressure ulcer, such as wherein said ulcer is a bed sore.

[0519] 117. The method for treatment according to item 108 or 109, wherein said wound is a boil, such as an open boil.

[0520] 118. The method for treatment according to any one of items 108 to 117, wherein said wound is an acute wound.

[0521] 119. The method for treatment according to any one of items 108 to 117, wherein said wound is a chronic wound.

[0522] 120. The method for treatment according to any one of items 108 to 119, wherein said wound is an infected wound.

[0523] 121. A method for tissue regeneration comprising administration of a composition as defined in any one of items 1 to 55 and 78, or a pharmaceutical formulation as defined in any one of items 79 to 81 , to a patient in need thereof.

[0524] 122. The method for treatment or for tissue regeneration according to any one of items 108 to 121 , wherein said administration is topical administration.

[0525] 123. The method for treatment or for tissue regeneration according to item 122, wherein said administration is a one-time administration.

[0526] 124. The method for treatment or for tissue regeneration according to item 122, wherein said administration is a repeated administration. 125. The method for treatment or for tissue regeneration according to any one of items 122 to 124, wherein said composition or said pharmaceutical formulation is in contact with said wound for at least approximately 8 hours, such as at least approximately 10 hours, such as at least approximately 12 hours, such as at least approximately 18 hours, such as at least approximately 24 hours, per administration.

[0527] 126. The method for treatment or for tissue regeneration according to any one of items 122 to 125, wherein said composition or said pharmaceutical formulation is in contact with said wound between 8 hours to 48 hours, such as between 12 hours to 48 hours, such as between 12 hours to 36 hours, such as between 12 hours to 24 hours, per administration.

[0528] 127. The method for treatment or for tissue regeneration according to any one of items 124 to 126, wherein there is a time interval between said repeated administrations, wherein said composition or said pharmaceutical formulation is not administered to said patient during said time interval.

[0529] 128. The method for treatment or for tissue regeneration according to item 127, wherein said time interval is at least 30 min, such as wherein said time interval is approximately 30 min.

[0530] 129. The method for treatment or for tissue regeneration according to item 127 or 128, wherein during said time interval a disinfection of the wound is performed.

[0531] 130. The method for treatment or for tissue regeneration according to any one of items 122 to 129, wherein a disinfection of the wound is performed prior to said administration.

[0532] 131 . The method for treatment or for tissue regeneration according to any one of items 122 to 130, wherein one or more disinfection of the wound is performed after a last administration of said composition or said pharmaceutical formulation.

[0533] 132. The method for treatment or for tissue regeneration according to any one of items 129 to 131 , wherein said wound is disinfected by using an oil, such as an oil derived from the species Pistacia atlantica, such as wherein the oil is derived from the seeds of the species Pistacia atlantica.

[0534] 133. The composition according to any one of items 1 to 55 and 78, or the pharmaceutical formulation according to in any one of items 79 to 81 , wherein said composition or said pharmaceutical formulation exhibits antimicrobial activity and / or anti-inflammatory activity.

[0535] 134. The composition or the pharmaceutical formulation according to item 133, wherein said antimicrobial activity is antibacterial activity.

[0536] 135. The composition according to any one of items 1 to 55 and 78, or the pharmaceutical formulation according to in any one of items 79 to 81 , wherein the weight ratio of said component of herb A, said component of herb B and said biomatrix of animal-origin is approximately 1 :1 :1 .5.

[0537] 136. The composition according to any one of items 1 to 55 and 78, or the pharmaceutical formulation according to in any one of items 79 to 81 , wherein said composition or said pharmaceutical formulation has tissue regenerative properties.

[0538] 137. The composition according to any one of items 1 to 55 and 78, wherein said composition consists of

[0539] - a component of herb A, wherein said herb A is selected from a species of the genus Pistacia',

[0540] - a component of herb B, wherein said herb B is selected from a species of the genus Salvia', and

[0541] - a biomatrix of animal-origin. 138. Use of

[0542] - a component of herb A, wherein said herb A is selected from a species of the genus Pistacia; - a component of herb B, wherein said herb B is selected from a species of the genus Salvia; and

[0543] - a biomatrix of animal-origin, in the manufacture of a medicament for the treatment for the treatment of wounds.

[0544] 139. Use of

[0545] - an herbal extract derived from a component of herb A, wherein said herb A is selected from a species of the genus Pistacia',

[0546] - an herbal extract derived from a component of herb B, wherein said herb B is selected from a species of the genus Salvia', and

[0547] - a biomatrix of animal-origin. in the manufacture of a medicament for the treatment for the treatment of wounds.

Claims

1. CLAIMS1 . A composition comprising- a component of herb A, wherein said herb A is selected from a species of the genus Pistacia',- a component of herb B, wherein said herb B is selected from a species of the genus Salvia', and- a biomatrix of animal-origin.

2. The composition according to claim 1 , wherein said herb A is of the species Pistacia atlantica.

3. The composition according to claim 1 or 2, wherein said component of herb A is an exudate of said herb A.

4. The composition according to claim 3, wherein said exudate of herb A is a resin or turpentine gum, such as turpentine gum.

5. The composition according to any one of the previous claims, wherein said herb B is of the species Salvia mirzayanii.

6. The composition according to any one of the previous claims, wherein said component of herb B is an element of the aerial subsystem of said herb B.

7. The composition according any one of the previous claims, wherein said component of herb B is a leaf.

8. The composition according to any one of the previous claims, wherein said biomatrix of animal-origin is a naturally occurring biological material.

9. The composition according to any one of claims 1 to 7, wherein said biomatrix of animal-origin is an artificially produced biological material.

10. The composition according to any one of the previous claims, wherein said biomatrix of animal-origin is an omentum and / or an equivalent thereof, such as wherein said biomatrix of animal-origin is an omentum.11 . The composition according to any one of the previous claims, wherein said biomatrix of animal-origin is derived from a mammal.

12. The composition according to claim 11 , wherein said biomatrix of animalorigin is derived from an animal of the family Bovidae, such as of the subfamily Caprinae, such as of the tribe Caprini.

13. The composition according to claim 11 or 12, wherein said biomatrix of animal-origin is derived from an animal of the species Capra aegagrus hircus.

14. The composition according to claim 11 , wherein said biomatrix of animalorigin is derived from a human.

15. A preparation method for a composition as defined in any one of the previous claims, wherein said method comprises the steps of(i) mixing of a component of herb A as defined in any one of claims 1 to 4; a component of herb B as defined in any one of claims 1 , 5 to 7; and a biomatrix of animal-origin as defined in any one of claims 1 , 8 to 14; wherein a mixture is obtained.

16. The preparation method according to claim 15, optionally further comprising at least one of the following steps (ii), (iii) and (iv):(ii) grinding of the mixture obtained in step (i); wherein a ground mixture is obtained;(iii) cooling of said mixture obtained in step (i) or of said ground mixture obtained in step (ii); and(iv) grinding of said mixture obtained in step (i) or of said ground mixture obtained in step (ii); or of the cooled mixture obtained in step (iii); wherein a reprocessed mixture is obtained.

17. The preparation method according to claim 15 or 16, wherein prior to step (i), said method comprises at least one step selected from the group consisting of:- freezing said component of herb A;- grinding of said component of herb A;- sieving of said component of herb A;- drying said component of herb B; and- freezing said biomatrix of animal-origin.

18. A pharmaceutical formulation comprising a composition as defined in any one of claims 1 to 14 comprising at least one pharmaceutically acceptable excipient.

19. The pharmaceutical formulation according to claim 18 formulated for topical administration.

20. A composition as defined in any one of claims 1 to 14, or a pharmaceutical formulation as defined in claim 18 or 19 for use as a medicament.21 . A composition as defined in any one of claims 1 to 14, or a pharmaceutical formulation as defined in claim 18 or 19 for use in the treatment of a wound, such as for use in wound healing.

22. The composition or the pharmaceutical formulation for use according to claim 21 , for use in promoting tissue regeneration and / or re-epithelialization of a wound.

23. The composition or the pharmaceutical formulation for use according to claim 21 or 22, wherein said wound is a penetrating wound, a boil or an ulcer.

24. The composition or the pharmaceutical formulation for use according to claim 23, wherein said penetrating wound is a bum.

25. The composition or the pharmaceutical formulation for use according to any one of claims 21 to 23, wherein said wound is an ulcer.

26. The composition or the pharmaceutical formulation for use according to claim 25, wherein said ulcer is selected from the group consisting of a peptic ulcer, a mouth ulcer, an arterial ulcer, a venous ulcer, a pressure ulcer, a diabetic ulcer and a genital ulcer.

27. The composition or the pharmaceutical formulation for use according to claim 25 or 26, wherein said ulcer is a diabetic ulcer.

28. A composition as defined in any one of claims 1 to 14, or a pharmaceutical formulation as defined in claim 18 or 19 for use in tissue regeneration, such as for use in tissue regeneration by re- epithelialization.

29. The composition or the pharmaceutical formulation for use according to claim 20 to 28, wherein said use comprises topical administration of said composition or of said pharmaceutical formulation to the wound of a patient in need thereof.

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