Dim compositions containing probiotic agent

Abstract

Provided herein, in some embodiments, is a composition for use in treating a skin condition, such as an inflammatory or infectious skin condition. In some embodiments, the composition comprises a first component comprising a substituted or unsubstituted diindolylmethane; and a second component comprising at least one probiotic agent, wherein the composition is essentially free of substituted or unsubstituted retinoid compounds, wherein the composition is essentially free of CYP450 enzyme modulators, and wherein the composition is essentially free of anti-protozoal agents.

Description

DIM COMPOSITIONS CONTAINING PROBIOTIC AGENTBACKGROUND OF THE DISCLOSURE

[0001] Diindoyly methane (DIM) is a component of Indole-3-carbinol (13 C) found in members of the Brassica family. Most notably broccoli, kale, and cauliflower.SUMMARY OF THE DISCLOSURE

[0002] Provided herein, in some embodiments, is a composition for use in treating a. skin condition, such as an inflammatory' or infectious skin condition. In some embodiments, the composition comprises a first component comprising a substituted or unsubstituted diindolylmethane; and a second component comprising at least one probiotic agent, wherein the composition is essentially free of substituted or unsubstituted, retinoid compounds, wherein the composition is essentially free of CYP450 enzyme modulators, wherein the composition is essentially free of antibiotic agents, and wherein the composition is essentially free of antiprotozoal agents.

[0003] In some embodiments, the additional therapeutic agent selected from one or more of, a substituted or unsubstituted azelaic acid compound, an oral contraceptive compound, sulphur, a sulphur-containing compound, a. substituted or unsubstituted salicylic acid compound, a substituted or un substituted resorcinol compound, a plant product, a mineral, a peroxide (such as benzoyl peroxide), a vitamin, and a neutraceutical product.

[0004] In some embodiments, the excluded substituted or unsubstituted retinoid compounds include substituted or unsubstituted first-generation retinoids, substituted or unsubstituted second- generation retinoids, and substituted or unsubstituted third-generation retinoids.

[0005] In some embodiments, the excluded substituted or unsubstituted first-generation retinoids include substituted or unsubstituted retinols, substituted or unsubstituted retinals, substituted or unsubstituted tretinoins, substituted or unsubstituted isoretinoins, and. substituted or unsubstituted alitretinoins.

[0006] In some embodiments, the excluded second-generation retinoids include substituted or unsubstituted etretinates and substituted or unsubstituted acitretins.

[0007] In some embodiments, the excluded third-generation retinoids include substituted or unsubstituted tazarotenes, substituted or unsubstituted bexarotenes, and substituted or unsubstituted adapalenes.

[0008] In some embodiments, the first component comprises a diindolylmethane of Formula 1:wherein the R groups are independently selected from hydrogen atoms and C1-C6hydrocarbon substituents; and wherein the indolyl groups are independently selected from indole-3 -yl and indole-2 -yl groups; and wherein the indolyl groups are unsubstituted, or are substituted with one or more C1-C6hydrocarbon substituents.

[0009] In some embodiments, the first component comprises a substituted or unsubstituted 3,3’diindolylmethane, of Formula 2, or a substituted or unsubstituted 2,2’diindolylmethane, of Formula 3:wherein the R groups are independently selected from hydrogen atoms, and C1-C6hydrocarbon substituents.

[0010] In some embodiments, the first component comprises a 3,3’diindolylmethane, of Formula 4 or a 2,2’diindolylmethane, of Formula 5:

[0011] In some embodiments, the first component comprises 3,3’diindolylmethane.

[0012] In some embodiments, the first component is in a form suitable for providing a. daily substituted or unsubstituted diindolylmethane dosage of up to 1500 mg or less. In some embodiments, the first component is in a form suitable for providing a daily substituted or unsubstituted diindolylmethane dosage of about 10 mg to about 750 mg.

[0013] In some embodiments, the substituted or unsubstituted diindolylmethane is adapted for increased bioavailability. In some embodiments, the substituted or unsubstituted diindolylmethane is microencapsulated. In some embodiments, the first component comprises one or more lipophilic compounds selected from vitamin E and phosphatidylcholine. In some embodiments, the substituted or unsubstituted diindolylmethane is BR-DIM.

[0014] In some embodiments, the composition is formulated for oral administration. In some embodiments, the composition is in the form of a tablet or capsule.

[0015] In some embodiments, the composition is formulated for topical administration. In some embodiments, the composition is in the form of a gel, cream, ointment, or a liquid.

[0016] In some embodiments, the first and second components are combined into one dosage form. In some embodiments, the first and. second components are in separate dosage forms.

[0017] In some embodiments, the first component is in a form suitable for providing a daily substituted or unsubstituted diindolylmethane dosage of about 10 mg to about 750 mg.

[0018] In some embodiments, the first component is in a. form suitable for providing a daily substituted or unsubstituted diindolylmethane dosage of about 150 mg to about 650 mg, about 250 mg to about 550 mg, about300 mg to about 500 mg, about 400 mg to about 450 mg, or about 500 mg.

[0019] In some embodiments, the first component is present in a dose which is a fraction of the daily dose, preferably a half of the daily dose, or a quarter of the daily dose.

[0020] In some embodiments, the composition is essentially free of any additional therapeutic agents other than DIM and probiotic agent.DESCRIPTION OF THE DRAWINGS

[0021] FIG. 1 includes photographs and Eventis redness images of a clinical study patient in DIM+Probiotic Group (Participant 2), showing noticeable improvements in redness.

[0022] FIG. 2 includes the photographs and Eventis redness images of a clinical study patient DIM+Probiotic Group (Participant 3), showing decreased inflammatory lesions and improvements in redness on the cheeks.

[0023] FIG. 3 includes the photographs and Eventis redness images of a clinical study patient DIM+Probiotic Group (Participant 4), showing noticeable improvements in redness.DETAILED DESCRIPTION OF THE DISCLOSURE

[0024] The present application discloses a pharmaceutical composition or kit comprising a substituted or unsubstituted, diindolylmethane (DIM) for use in treating skin diseases or conditions. In some embodiments, the composition comprises one or more other components in addition to the substituted or unsubstituted DIM in order to enhance the pharmaceutical effect of the substituted or unsubstituted DIM.Certain Definitions

[0025] Unless defined otherwise, ail technical and scientific terms used herein have the same meaning as is commonly understood by one of skill in the art to which the claimed subject matter belongs. In the event that there is a plurality of definitions for terms herein, those in this section prevail.

[0026] It is to be understood that the foregoing general description and the following detailed description are exemplary and explanatory' only and are not restrictive of any subject matter claimed. In this application, the use of the singular includes the plural unless specifically stated otherwise. It must be noted that, as used in the specification and the appended claims, the singular forms "a", "an" and "the" include plural referents unless the context clearly dictates otherwise. It should also be noted that use of "or" means "and / or" unless stated otherwise. Furthermore, use of the term "including" as well as other forms, such as "include", "includes", and "included" is not limiting.

[0027] As used herein, the term “about” is used synonymously with the term “approximately.” Illustratively, the use of the term “about” with regard to a certain therapeutically effective pharmaceutical dose indicates that values slightly outside the cited values, e.g., plus or minus 0.1 % to 10%, are also effective and safe.

[0028] The term "patient", "subject" or "individual" are used interchangeably. As used herein, they refer to individuals suffering from a disorder, and the like, encompasses mammals and nonmammals. None of the terms require that the individual be under the care and / or s upervision of a medical professional. Mammals are any member of the Mammalian class, including but not limited to humans, non-human primates such as chimpanzees, and other apes and monkey species; farm animals such as cattle, horses, sheep, goats, swine; domestic animals such as rabbits, dogs, and cats; laboratory' animals including rodents, such as rats, mice and guinea pigs, and the like. Examples of non-mammals include, but are not limited, to, birds, fish and the like. In someembodiments of the methods and compositions provided herein, the individual is a mammal. In preferred embodiments, the individual is a human.

[0029] The terms "treat," "treating" or "treatment," and other grammatical equivalents as used herein, include alleviating, abating or ameliorating a. disease or condition or one or more symptoms thereof, preventing additional symptoms, ameliorating or preventing the underlying metabolic causes of symptoms, inhibiting the disease or condition, e.g., arresting the development of the disease or condition, relieving the disease or condition, causing regression of the disease or condition, relieving a condition caused by the disease or condition, or stopping the symptoms of the disease or condition, and. are intended to include prophylaxis. The terms further include achieving a therapeutic benefit and / or a prophylactic benefit. By therapeutic benefit is meant eradication or amelioration of the underlying disorder being treated. Also, a. therapeutic benefit is achieved, with the eradication or amelioration of one or more of the physiological symptoms associated with the underlying disorder such that an improvement is observed in the individual, notwithstanding that the individual is still be afflicted with the underlying disorder. For prophylactic benefit, the compositions are administered to an individual at risk of developing a. particular disease, or to an individual reporting one or more of the physiological symptoms of a. disease, even though a diagnosis of this disease has not been made.

[0030] The terms "administer," "administering”, "administration," and the like, as used herein, refer to the methods that may be used to enable deliver}' of compounds or compositions to the desired site of biological action. These methods include, but are not limited to topical routes, oral routes, intraduodenal routes, parenteral injection (including intravenous, subcutaneous, intraperitoneal, intramuscular, intravascular or infusion), topical and rectal administration. Those of skill in the art are familiar with administration techniques that can be employed with the compounds and methods described herein.

[0031] The term "acceptable" as used herein, with respect to a formulation, composition or ingredient, means having no persistent detrimental effect on the general health of the individual being treated.

[0032] Tire term "pharmaceutically acceptable" as used herein, refers to a material, such as a carrier or diluent, which does not abrogate the biological activity or properties of the compounds described herein, and is relatively nontoxic, i.e., the material may be administered to an individual without causing undesirable biological effects or interacting in a deleterious manner with any of the components of the composition in which it is contained.

[0033] Die term “DIM” as used herein, refers to a. substituted or unsubstituted diindolylmethane compound.

[0034] The term “BR-DIM or “BioResponse DIM” as used herein, refers to an unsubstituted DIM, provided by BioResponse LLC.

[0035] The term “maximum concentration” or Cmax as used herein, refers to the Cmax refers to the maximum (or peak) serum concentration that the substituted or unsubstituted diindolylmethane achieves in the plasma after it has been administrated and prior to the administration of a second dose.

[0036] The term “time to maximum concentration” or Tmax as used herein, refers to the time at which the Cmax is observed.

[0037] The term “area under the curve” or AUC or AUCo-inf as used herein, refers to the area under the curve, also known as the definite integral, in a. plot of concentration of drug in blood plasma against time.

[0038] The term “unit dose” as used herein, refers to an amount of substituted or unsubstituted diindolylmethane contained in one discreet pharmaceutical dosage form. Examples of pharmaceutical dosage form that contains a unit dose include but are not limited to a tablet, a. capsule, a buccal tablet, a sub-lingual tablet, an orally-disintegrating tablet, an effervescent tablet, a. lollipop, a lozenge, a troche, a liquid solution or suspension, powder or liquid or solid crystals packed within a single tablet or capsule, a cream, a. gel, an ointment, a lotion.

[0039] DIM

[0040] As such, provided herein, in one embodiment, are methods and compositions for use in treating skin conditions, such as inflammatory skin conditions, wherein the composition comprises a first component, optionally a second component, wherein the first component comprises a substituted or unsubstituted diindolylmethane (DIM), and the second component comprises a substituted or unsubstituted retinoid compound. In some embodiments, are provided kits containing the compositions for use in treating skin conditions, such as inflammatoiy skin conditions, as described herein. In some embodiments, the DIM is a substituted or an unsubstituted DIM. In some embodiments, the retinoid compound is a substituted or an unsubstituted retinoid compound. In some embodiments, the DIM is BioResponse DIM (BR-DIM). In some embodiments, the DIM is any compound comprising a diindolylmethane group. In some embodiments, the DIM has the following structure (Formula 1):wherein the R groups are be the same or different substituents. In some embodiments, the R substituents comprise any organic group and / or one or more atoms from any of groups IIIA, IVA, VA, VIA or VILA of the Periodic Table, such as a B, Si, N, P, O, or S atom or a halogen atom (e.g, F, Cl, Br or I).

[0041] In some embodiments, the R substituent comprises an organic group. In some embodiments, the organic group comprises a hydrocarbon group. In some embodiments, the hydrocarbon group comprises a straight chain, a branched chain or a cyclic group. In some additional embodiments, the hydrocarbon group comprises an aliphatic or an aromatic group. In some additional embodiments, the hydrocarbon group comprises a saturated or an unsaturated group.

[0042] In some embodiments, the hydrocarbon comprises an unsaturated group comprising one or more alkene functionalities and / or one or more alkyne functionalities. In some embodiments, the hydrocarbon comprises a straight or branched chain group comprising one or more primary, secondary and / or tertian,7alkyl groups. In some embodiment, the number of carbon atoms in the hydrocarbon group is between 1-40. In some embodiments, the hydrocarbon group is a lower hydrocarbon. In some embodiments, the number of carbon atom in the lower hydrocarbon is between 1 and 6. In some additional embodiments, the hydrocarbon group is a higher hydrocarbon. In some embodiments, the number of hydrocarbons in the higher hydrocarbon is higher than 7. In some embodiments, the number of carbon atoms in the higher hydrocarbon is between 7 and 40.

[0043] In some embodiments, the hydrocarbon comprises a cyclic group comprising an aromatic ring, an aliphatic ring, a heterocyclic group, and / or fused ring derivatives of these groups. In some embodiments, the cyclic group comprises a benzene, a naphthalene, an anthracene, an indene, a fluorene, a pyridine, a quinoline, a thiophene, a benzothiophene, a furan, a benzofuran, a pyrrole, an indole, an imidazole, a thiazole, and / or an oxazole group, as well as regioisomers of the above groups. In some embodiments, the number of atoms in the ring of the cyclic group is between 3 and 10. In some embodiments, the number of atoms in the ring of the cyclic group is 3, 4, 5, 6, or 7.

[0044] In some embodiments, the cyclic groups comprising heteroatoms described above, as well as any of the other groups defined above, further comprise one or more heteroatoms from any ofgroups IIIA, IVA, VA, VIA or VILA of the Periodic Table, such as a B, Si, N, P, O, or S atom or a halogen atom (e.g.. F, Cl, Br or I).

[0045] In some embodiments, the R substituent comprises one or more of any of the common functional groups in organic chemistry, such as hydroxy groups, carboxylic acid groups, ester groups, ether groups, aldehyde groups, ketone groups, amine groups, amide groups, imine groups, thiol groups, thioether groups, sulphate groups, sulphonic acid groups, and phosphate groups etc. In some embodiments, the substituent comprises one or more derivati ves of any of the common functional groups in organic chemistry, such as hydroxy groups, carboxylic acid groups, ester groups, ether groups, aldehyde groups, ketone groups, anime groups, amide groups, imine groups, thiol groups, thioether groups, sulphate groups, sulphonic acid groups, and phosphate groups etc. Exemplary derivatives include but are not limited to, carboxylic acid anhydrydes and carboxylic acid halides.

[0046] In some embodiments, any R substituent comprises a combination of two or more of the substituents and / or functional groups defined above.

[0047] In some embodiments, the R substituents are selected from hydrogen atoms and C1-C6hydrocarbon substituents such as C1-C6alkyl groups (e.g,. methyl, ethyl, propyl, isopropyl, and butyl groups). In some embodiments, both R substituents are hydrogen atoms.

[0048] In some embodiments, the indolyl groups in formula 1 are the same or different. In some embodiments, both indolyl groups are indole-3-yl groups. In some embodiments, both indolyl groups are indole-2-yl groups. In some embodiments, one indolyl group is indole-3-yl and the other indolyl group is indole-2-yl. In some embodiments, the indolyl groups comprise same or different substituents. In some embodiments, the indolyl groups are unsubstituted, such that all substituents are hydrogen atoms. In some embodiments, the indolyl groups are substituted with one or more of any of the R substituents defined above. In some embodiments, the substituents are selected from hydrogen atoms and C1-C6hydrocarbon substituents such as C1-C6alkyl groups (e.g, methyl, ethyl, propyl, isopropyl, and butyl groups).

[0049] In some embodiments, the composition comprises a first component comprising a substituted or unsubstituted 3,3’diindolylmethane, of Formula 2. In some embodiments, the composition comprises aa first component comprising a a. substituted or unsubstituted 2,2’diindolylmethane, of Formula 3.wherein the R groups are independently selected from hydrogen atoms, or C1-C.5 hydrocarbon substituents.

[0050] In some embodiments, the composition comprises a first component comprising a 3,3’drindolylmethane, of Formula 4. In some embodiments, the composition comprises a first component comprising a 2,2’diindolylmethane, of Formula 5.Probiotic gents

[0051] In some embodiments, the second component has a beneficial effect supplementary’ to and / or complementary to the effect of the first component. In some embodiments, the compositions described herein comprise probiotic agents. The probiotic agents can be pre-biotic agents or post-biotic agents. The probiotic agents include, but are not limited to. Lactobacillus rhamnosus, Lactobacillus paracasei. Lactobacillus acidophilis, Lactobacillus salivarius. Lactobacillus sakei. Lactobacillus johnsonii, Lactobacillus reuteri, Lactobacillus bulgaricus. Lactobacillus plantarum. Lactobacillus sporogenes, Bifidobacterium lactis, Bifidobacterium breve, Bifidobacterium longum, Bifidobacterium bifidum. Bifidobacterium infantis, Roseomonas mucosa, Propriombactenum, Vitreoscilla filiformis, Streptococcus thermophilus, and Enterococcus faecalis.

[0052] In some embodiments, the composition described herein comprises Lactobacillus paracasei and Saccharomyces boulardii. Such a bacteria and. yeast may be used to improve the appearance of the skin, normalise the concentration of sebum on the skin, and may be used to improve the appearance of inflammatory skin diseases or even treat such diseases. A composition comprising Lactobacillus paracasei and Saccharomyces boulardii may be particularly useful in reducing the appearance of, or treating inflammatory skin diseases such as acne vulgaris, atopic dermatitis, psoriasis and rosacea.

[0053] Preferably, the Lactobacillus paracasei comprises the strain Lafti L26 AF, and the Saccharomyces boulardii comprises the strain Saccharomyces cerevisiae boulardii CNCM -I- 1079. These specific strains of Lactobacillus paracasei and Saccharomyces boulardii may be particularly advantageous for improving the appearance of the skin or treating an inflammatory' skin disease over other strains of Lactobacillus paracasei and Saccharomyces boulardii.

[0054] Preferably, the composition at the point of manufacture comprises between about 1 x 109and about 20 x 109colony forming units (CFUs) of Lactobacillus paracasei and between about 1 x IO9and about 4 x IO9of Saccharomyces boulardii, preferably wherein the composition comprises about 12 x 109or more colony forming units (CFUs) of Lactobacillus paracasei and about 2.5 x 109CFU or more of Saccharomyces boulardii. Such a concentration of each of Lactobacillus paracasei and Saccharomyces boulardii may advantageously result in the improvement of the appearance of the skin or treatment of an inflammatory' skin disease of the skin.

[0055] Preferably, the composition according to the first aspect of the invention further comprises Lactobacillus helve ticus, preferably wherein the Lactobacillus helveticus comprises the strain Lafti LI O ND. The presence of Lactobacillus helveticus in the composition may further improve the appearance of the skin of a subject that has orally been administered the composition or may treat inflammatory skin diseases. Preferably, the composition comprises at the point of manufacture between about 1 x 109and about 8 x 109colony' forming units (CFUs) of Lactobacillus helveticus, preferably wherein the composition comprises about 4 x 109or more colony forming units (CFUs) of Lactobacillus helveticus.Preferably, the composition according to the first aspect of the invention further comprises further comprising Bifidobacterium bifldum, preferably' wherein the Bifidobacterium bifidum comprises the strain Rosell-71 (R0071 ). Preferably, the composition comprises at the point of manufacture between about 1 x 109and about 8x 109colony forming units (CPUs) of 'Bifidobacterium bifidum, preferably wherein the composition comprises about 4 x 109or more colony forming units (CPUs) of Bifidobacterium bifidum.

[0056] Preferably, the total amount of Lactobacillus paracasei and Saccharomyces boulardii, and optionally each of Lactobacillus helveticus and Bifidobacterium bifidum is at least 5 x 109colony forming units at the point of oral administration of the composition. Due to the fact that bacteria will naturally die-off once the composition has been formulated, the composition contains at least at least 5 x 10 |9 colony forming units of all yeast and bacteria strains at the point of oral administration. This may alternatively be measured as a concentration of at least at least 5 x 109colony forming units of all yeast and bacteria strains at the best before date of the composition i.e. the date at which at any later date the composition is deemed no longer fit for human use.Microbial Strains

[0057] Compositions according to the invention may in certain embodiments comprise one or more preferred strains as shown in the table below.

[0058] According to certain embodiments, one or more of the microbial species may in an optional embodiment comprise an equivalent, corresponding, similar or derivative of the strain listed above. For example, it may comprise a redeposit of one of the strains listed above. It may be a strain winch is isolated from the same or similar source as one of the strains listed above, or it may be a strain for which one of the strains listed above is an ancestral strain, that is to say it may be a strain winch was been obtained by natural or artificial means from one of the specified strains. Form of microbes

[0059] In preferred embodiments the microbes specified as part of the various aspects of the invention are provided in a viable or substantially viable form. Typically this may be in a freeze dried or otherwise preserved form in which metabolic activity islow but wherein a material proportion of the cells retain the ability to increase their metabolic activity' and optionally multiply under appropriate conditions. Level of specified microbial organisms

[0060] The number of viable cells of each of the specified microbial species is conveniently expressed in terms of colony -forming units (CPUs). This may be assessed by any appropriate method. For example as outlined in Goldman, Emanuel; Green, Lorrence H Practical Handbook of Microbiology, Second Edition (Second ed.). USA: CRC Press, Taylor and Francis Group, p. 864. ISBN 978-0-8493-9365-5 (herein incorporated by reference).

[0061] According to certain embodiments of the invention the total amount of bacteria, present in a composition according to the invention is about 20 x 109CFU at the point of manufacture. In some embodiments, the total amount of Lactobacillus paracasei in a composition of the invention may- be between about 1 x 109and about 20 x 109CFU, and optionally7the total amount of Bifidobacterium bifidum may be between about 1 x 109and about 19 x 109CFU if present in the composition, and optionally the total amount of Lactobacillus helveticus may be between about 1 x 10 =9 and about 19 x 109CFU if present in the composition. In such embodiments, if the composition comprises two or more of 'Lactobacillus paracasei. Bifidobacterium bifidum and Lactobacillus helveticus, the combined total number of CFU of all bacterial species may not exceed 20 x 109CFU. For example, the total amount of Bifidobacterium bifidum may7be 4 x 109CFU, the total amount of Lactobacillus paracasei may be 12 x IO9CFU, and the total amount of Lactobacillus helveticus may be 4 x ID9CFU, at the point of manufacture.

[0062] It is likely' that in most compositions there will also be a certain level of nonviable cells and / or cell debris. However according to certain embodiments, viable microbial cells in total make up at least 5, 10, 20, 30, 40, 50, 60, or 70 % of the total weight of the composition.

[0063] After the point of manufacture, the amount of bacteria present in the composition will decrease owing to die-off A convenient way of expressing this is by determining the minimum amount of bacteria, present at the best before date or the minimum amount of bacteria present at the date the composition is orally administered. According to certain embodiments the total amount of Lactobacillus paracasei and Saccharomyces boulard.it, and optionally each of Lactobacillushelveticus and Bijidobacterium bifldum, in the composition at the point of oral administration and / or at the best before date of the composition is at least 2 x IO9CPUs, at least 5 x 109CPUs, or as much as 10 x 109CPUs, at the point of oral administration of the composition.Excluded Therapeutic Agents

[0064] In some embodiments, the compositions described herein is essentially free of any substituted or unsubstituted retinoid compound. In some embodiments, the excluded retinoid compound is any such compound known in the art that is suitable for use with the skin. For example, in some embodiments, it is selected from a substituted or unsubstituted first generation retinoid, a substituted or unsubstituted second generation retinoid, and a substituted or unsubstituted third generation retinoid. In some embodiments, the retinoid is a substituted or unsubstituted first generation retinoid. In some embodiments, the substituted or unsubstituted first generation retinoid is selected from a substituted or unsubstituted retinol, a substituted or unsubstituted retinal, a substituted or unsubstituted tretinoin (e.g., retinoic acid or Retin A), a substituted or unsubstituted isotretinoin (e.g,. AccutaneTM), and a substituted or unsubstituted alitretinoin. In some embodiments, the retinoid is vitamin A. In some embodiments, the retinoid is a substituted or unsubstituted second generation retinoid selected from a substituted or unsubstituted etretinate, and a substituted or unsubstituted acitretin. In some embodiments, the retinoid is a substituted or unsubstituted, third generate on retinoid selected from a substituted or unsubstituted tazarotene, a substituted or unsubstituted bexarotene, and a substituted or unsubstituted adapalene. In some embodiments, the excluded retinoid compound is a vitamin A compound (e.g, vitamin A palmitate).

[0065] In some embodiments, the compositions described herein is essentially free of CYP450 enzyme modulators. In some embodiments, the excluded CYP450 enzyme modulator is any such compound known in the art that is suitable for use with the skin. For example, in some embodiments, it is selected from CYP1A1, C YP1A2, CYP2D6, CYP2C8, CYP2C9, CYP3A4, CYP2CT9, and CYP19A1.

[0066] In some embodiments, the compositions described herein is essentially free of antibiotic agents. The excluded antibiotic agents include, but are not limited to, lincosamides (such as clindamycin), sulfones (such as dapsone), tetracyclines (such as tetracycline, doxycycline, minocycline, iymecyciine), macrolides (such as erythromycin, azithromycin), sulphonamides (such as sulfamethoxazole, co-trimoxazole), diaminopyrimidines (trimethoprim), penicillin and its derivatives, and cephalosporins and its derivatives,

[0067] In some embodiments, the compositions described herein is essentially free of antiprotozoal agents. In some embodiments, the excluded anti -protozoal agent is any such compound known in the art that is suitable for use with the skin. For example, in some embodiments, it is selected from atovaquone, amodiaquine, amphotericin, butoconazole, clindamycin, eflomithine, fumagillin, iodoquinol (diiodohydroxy quin), clioquinol (iodochlorhydroxyquin), Etanidazole, Benzmdazole, fluoroquinolones, enoxacin, ciprofloxacin, doxycycline, melarsoprol, metronidazole, miltefosine, nifurtimox, nitazoxanide, paromomycin, pentamindine, sodium stibogluconate, suramin, tinidozole, pyrimethamine, proguaml (chloroguanide), spiramycin, and sulfadoxine. Natural product derived antiprotozoal drugs useful for combined use include sesquiterpene lactones related to artemisinin from Artemisia annua, particularly artemisinin, dihydroartemisinin, artemether, artesunate, and further derivatives of artemisinin, quinolines like quinine derived from the bark of the South American chinchona tree, including quinine and quinine-related quinolines, halofantrine, mefloquine, lumefantrine, amodiaquine, pyronaridine, prperaquine, chloroquine, hydroxychloroquine, napthoquine, primaquine, and tafenoquine, curcummoids derived from curcumin, an extract from Curcuma domestica, including 6-gingerol and 6-paradol, coronaridine, 18- methoxycoronaridine, selected flavonoids, including luteolin, extracts the fruit pericarp of Sapindus mukorossi, and extracts of Yucca schidigera. Excluded antiprotozoal agent also include artemisinin derivatives, atovaquone, chloroquine, iodoquinol (diiodohydroxy quin), clioquinol (iodochlorhydroxy' quin), sodium stibogluconate, and curcumin.. Excluded antiprotozoal agent also include antiprotozoal natural products such as teas and extracts made from Artemisia, annua, teas and extracts made from Curcuma domestica, extracts from garlic which include allicin and other thiosulfinates, root extracts of Uvana chamae (Annonaceae) and Hippocratea Africana (Hippocrateaceae), and root extracts of Homalium letestui.Bioavailability of substituted or unsubstituted DIM

[0068] Dimdolylmethane (DIM) is a natural compound, formed during the autolytic breakdown of glucobrassicin present in food plants of the Brassica genus, including broccoli, cabbage, Brussels sprouts, cauliflower and kale. The autolytic breakdown of glucobrassicin requires the catalytic reaction of the enzyme myrosinase, which is endogenous to these plants and released upon rupture of the cell w all. The compound is normally manufactured by chemical synthesis but in some embodiments is also prepared by natural means from the extracts of Brassica vegetables, as listed above, particularly from sprouting broccoli or from broccoli seeds. Thus, the substituted or unsubstituted DIM in some embodiments is synthetic, or in someembodiments is a. natural product obtained from a. Brassica plant, as discussed above. In some embodiments, the composition described herein, includes as a first component, any substituted, or unsubstituted DIM, or a combination of the substituted or unsubstituted DIMs described above.

[0069] In some embodiments, the substituted or unsubstituted DIM is adapted for increased bioavailability, in order to reduce the required dosage. In some embodiments, the DIM with increased bioavailability is an unsubstituted DIM. In some embodiments, the DIM with increased bioavailability is BR-DIM (from BioResponse LLC).Dosage of administration

[0070] In some embodiments, the composition comprising a first component comprising a substituted or unsubstituted DIM, is administered at a dosage that is sufficiently low to avoid toxicity, whilst still maintaining the required pharmaceutical effect. In some embodiments, the dosage depends on the bioavailabihty of the substituted or unsubstituted DIM. In some embodiments, the dosage varies depending upon whether it is a natural or synthetic product. In some embodiments, the dosage of the first component comprising the substituted or unsubstituted. DIM is determined by whether or not the DIM has been adapted to improve bioavailabilitw

[0071] In some embodiments, the first component comprising a substituted or unsubstituted DIM is in a form suitable for providing a daily DIM dosage of 1500 mg or less, 1000 mg or less, 900 nig or less, 800 mg or less, 700 mg or less, 600 mg or less, 500 mg or less, from 10-750 mg, from 150-650 mg, from 250-550 mg, and from 300-500 mg. In some embodiments, the first component comprising a. substituted or unsubstituted DIM is in a form suitable for providing a. daily DIM dosage of about 300 mg or about 350 mg, or about 400 mg or about 450 mg, or about 500 mg. In some embodiments, the first component comprising a BR-DIM is in a form suitable for providing a daily BR-DIM dosage of 1500 mg or less, 1000 mg or less, 900 mg or less, 800 mg or less, 700 mg or less, 600 mg or less, 500 mg or less, from 10-750 mg, from 150-650 mg, from 250-550 mg, and from 300-500 mg. In some embodiments, the first component comprising a BR-DIM is in a form suitable for providing a daily BR-DIM dosage of about 300 mg or about 350 mg, or about 400 mg or about 450 mg, or about 500 mg. In some embodiments, the first component comprising a BR-DIM that has not been adapted to improve bioavailabihty, is administered in higher dosages.

[0072] In some embodiments, the first component comprising a substituted or unsubstituted DIM is in a form suitable for providing an intermediate daily DIM dosage, such as from 50-350 mg, 100-200 mg, 140-160 mg, or about 150 mg, or alternatively 250-350 mg, 290-310 mg or about 300 mg. In some embodiments, the first component comprising a substituted or unsubstituted DIM is in a form suitable for providing a low daily DIM dosage, such as from 15-100 mg, 50-100 mg, 18-80 mg, 18-75 mg, 70-80 mg, or about 75 mg. In some embodiments, the first component comprising a BR-DIM is in a form suitable for providing an intermediate daily BR-DIM dosage, such as from 50-350 mg, 100-200 mg, 140-160 mg, or about 150 mg, or alternatively 250-350 mg, 290-310 mg or about 300 mg. In some embodiments, the first component comprising a. BR-DIM is in a form suitable for providing a low daily BR-DIM dosage, such as from 15-100 mg, 50-100 mg, 18-80 mg, 18-75 mg, 70-80 mg, or about 75 mg.

[0073] In some embodiments, the first component comprising a substituted or unsubstituted DIM that has been adapted to improve bioavail ability is administered in intermediate and lower dosages. In some embodiments, the first component comprising a substituted or unsubstituted DIM that has been adapted to improve bioavailability is administered in any of the above dosages, including the higher dosages, if desired. In some embodiments, the DIM that has been adapted to improve bioavailability is BR-DIM.

[0074] In some other embodiments, the first component comprising a substituted or unsubstituted DIM is present in a dose which is a fraction of the daily dose, such as a half of the daily dose, or a quarter of the daily dose, and thus is present in a half or a quarter of any of the dosages recited above. In these embodiments, each dose fraction is taken separately over time to spread the dose across the day. In some other embodiments, the first component comprising a BR-DIM is present in a dose which is a fraction of the daily dose, such as a half of the daily dose, or a quarter of the daily dose, and thus is present in a haff or a. quarter of any of the dosages recited above. In these embodiments, each dose fraction is taken separately over time to spread the dose across the day.

[0075] In some embodiments, the second component comprising an additional therapeutic agent is administered at a. dosage that it is sufficiently low to avoid, toxicity, whilst still maintaining the required pharmaceutical effect. In some embodiments, the dosage of the second component depends on the bioavailability of the retinoid compound. In some embodiments, the bioavailability of the additional therapeutic agent, varies depending upon whether it is a natural or synthetic product. In some embodiments, the bioavailability of the additional therapeutic agent, may vary depending on whether it has been adapted to improve its bioavailability. In some embodiments, the second component comprising an additional therapeutic agent is in a form suitable for providing a daily dosage of 250 mg or less, 200 mg or less, 150 mg or less,100 mg or less, 50 mg or less, or 15 mg or less. In some embodiments, the second component comprising an additional therapeutic agent is in a form suitable for providing a. daily dosage of 10,000 μg or less, 5,000 μg or less, or 3,000 μg or less. In some embodiments, the second component comprising an additional therapeutic agent is in a. form suitable for providing a dailydosage of 50 μg or more, 100 μg or more, or 200 μg or more. In some embodiments, the second, component comprising an additional therapeutic agent is in a form suitable for providing a dailydosage from 200-15,000 μg, from 200-5000 μg, or from 200-3000 μg.

[0076] In some embodiments, daily dosage of the composition comprising a first component and a second, component is provided in the form of one or more capsules or tablets. In some embodiments, daily dosage of the composition comprising a first component and a second component is provided in the form of 2 or 4 capsules or tablets. In these embodiments the capsules or tablets are taken during the course of a. single day, such as one in the morning and one in the evening, or four spread evenly across the day, or two capsules or tablets simultaneously twice a day.Dosage Form

[0077] In some embodiments, the compositions described herein are used to prepare a formulation suitable for administration to a human or other subject. In some embodiments, the composition components are formulated for oral or topical administration. In some embodiments, the composition components, are provided in the form of a tablet, capsule, liquid (such as in the form of a drink, e.g., a yoghurt drink), gel, cream, or ointment. In some embodiment, the composition components comprise the first component present in a first formulation, and the optional second component present in a second formulation, wherein the first and. second formulations are co-administered to a patient, either concurrently or in a sequential manner. In some embodiment, the composition and / or the kit components comprises the first component and second component present in one formulation that is administered to a patient.Methods of Preparation

[0078] In some embodiments, are provided, methods for preparing the compositions described herein, suitable for use in treatment of skin conditions, such as inflammatory skin conditions. In some embodiments, any methods known in the art for blending or mixing various components of the composition are employed. In some embodiments, the methods employed are methods for blending and / or mixing powders. In some embodiments, the method comprises mixing a first component with one or more excipients and / or additives, optionally with a second component.and optionally a further component to form the composition. In some embodiments, the first, second, and / or further components are each, separately from each other, mixed with one or more excipients anchor additives before being mixed together to form the composition. In some additional embodiments, the components, additives, and / or excipients are added sequentially to the mixture during the mixing process.

[0079] In some embodiments, the selection of the excipients and the method of blending are adapted in order to overcome any mixing, flow and fill issues or punch issues with the composition. In some embodiments, the composition comprising a first component comprising a substituted or unsubstituted DIM or a. BR-DIM, is provided in micro-encapsulated form, such that the powder particles have a tendency to clump together. In some embodiments, the composition comprising a first component, comprising a substituted or un substituted DIM or a. BR-DIM is blended using a method that is adapted to avoid creating hot spots of increased concentrations of the active ingredients. In some embodiments, the composition comprising a first component, optionally a second component, and optionally a further component, is blended using a method that involves short processing / blending times, to protect the composition from light and air, wherein the composition is hygroscopic and light sensitive. In some embodiments, the composition comprising a first component, optionally a second component, and optionally a further component, is prepared in the form of a. powder, and the powder is protected from both light and air, during storage.

[0080] In some embodiment, one or more of microcrystalline cellulose, magnesium silicate, tricalcium phosphate, and magnesium stearate (a traditional lubricant) are employed as additives and excipients, in preparing the compositions described herein, to help with flow characteristics and / or lubrication. In some embodiments, other additives and excipients known in the art are employed if desired. In some embodiments, the composition comprises 50.0-65.0% by weight of tri-calcium phosphate. In some embodiments, the composition comprises, 55.0-60.0% by weight, or 57.0-59.0% by weight of tri-calcium phosphate. In some embodiments, the composition comprises about 58% by weight of tri-calcium phosphate. In some embodiments, the composition comprises about 58.3% by weight of tri-calcium phosphate.

[0081] In some embodiments, the correct blending of all ingredients is desirable in achieving uniform capsule fills of the compositions as described herein. In some embodiments, the correct blending of all ingredients is desirable in achieving uniform capsule fills of the compositions as described herein. In some embodiments, a V -blender is highly effective for successful mixing. In some embodiments, a. minimum 316 grade stainless steel vessel is used, for the mixingprocess. In some embodiments, sieving is performed at one or more of the start, the middle, and the end of the mixing process. In some embodiments, blend studies to confirm blend uniformity are completed to validate the method and formulation, using methods and techniques known in the field.NON-LIMITING EMBODIMENTS

[0082] The following specific, non-limiting embodiments are to be construed as merely illustrative, and do not limit the present disclosure of the scope of the disclosure. Without further elaboration, it is believed that one skilled in the art can, based on the description herein, utilize the present disclosure to its fullest extent.1. A composition for use in treating a skin disease or condition, the composition comprising: a first component comprising a substituted or unsubstituted diindolylmethane: and a second component comprising at least one probiotic, agent, wherein the composition is essentially free of substituted or unsubstituted retinoid compounds, wherein the composition is essentially free of CYP450 enzyme modulators, and wherein the composition is essentially free of anti-protozoal agents.2. The composition of embodiment 1, wherein the composition further comprises a further component selected from one or more of, a substituted or unsubstituted azelaic acid compound, an oral contraceptive compound, sulphur, a sulphur-containing compound, a substituted or unsubstituted salicylic acid compound, a substituted or unsubstituted resorcinol compound, a peroxide (optionally benzoyl peroxide), a. plant product, a mineral, a vitamin, and a neutraceutical product.3. The composition of embodiment 1-2, wherein the substituted or unsubstituted retinoid compounds comprise substituted or unsubstituted first-generation retinoids, substituted or unsubstituted second-generation retinoids, and substituted or unsubstituted third-generation retinoids.4. The composition according to embodiment 3, wherein the substituted or unsubstituted first-generation retinoids comprise substituted or unsubstituted retinols, substituted or unsubstituted retinals, substituted or unsubstituted tretinoins, substituted or unsubstituted isoretinoins, and substituted or unsubstituted alitretinoins.5. The composition according to embodiment 3, wherein the second- generation retinoids comprise substituted or unsubstituted etretinates and substituted or unsubstituted acitretins.6. The composition according to embodiment 3, wherein the third-generation retinoids comprise substituted or unsubstituted tazarotenes, substituted or unsubstituted bexarotenes, and substituted or unsubstituted adapalenes.7. The composition of embodiment 1-6, wherein the first component comprises a dimdolylmethane of Formula 1:wherein the R groups are independently selected, from hydrogen atoms and C1-C6hydrocarbon substituents; and wherein the indolyi groups are independently selected from indole-3-yl and indole-2-yl groups; and wherein the indolyi groups are unsubstituted, or are substituted with one or more C1-C6hydrocarbon substituents.8. The composition of embodiment. 1-6, wherein the first component comprises a substituted or unsubstituted 3,3’diindolylmethane, of Formula 2, or a. substituted or unsubstituted 2,2 / dimdolylmethane, of Formula 3:wherein the R groups are independently selected from hydrogen atoms, and C1-C6hydrocarbon substituents.9. The composition of embodiment 1-6, wherein the first component comprises a 3,3’diindolylmethane, of Formula 4 or a 2,2’diindolylmethane, of Formula 5:10. The composition of embodiment 1-6, wherein the first component compri s es 3 , 3 ’ dii ndoly 1 meth ane.11. The composition of embodiment 1-10, wherein the first component is in a form suitable for providing a daily substituted or unsubstituted diindolylmethane dosage of up to 1500 mg or less.12. The composition of embodiment 1-10, wherein the first component is in a form suitable for providing a daily substituted or unsubstituted diindolylmethane dosage of about 10 mg to about 750 mg.13. The composition of embodiment 1 -12, wherein the substituted or unsubstituted diindolylmethane is adapted for increased bioavailability.14. The composition of embodiment 13, wherein the substituted or unsubstituted, diindolylmethane is microencapsulated.15. The composition of embodiment 14, wherein the first component comprises one or more lipophilic compounds selected from vitamin E and phosphati dylchohne.16. The composition of embodiment 13, wherein the substituted or unsubstituted diindolylmethane is BR-DIM.17. The composition of embodiment 1-16, wherein the composition is formulated for oral administration.18. The composition according to embodiment 17, wherein the composition is in the form of a tablet or capsule.19. The composition of embodiment 1-16, wherein the composition is formulated, for topical administration.20. The composition of embodiment 19, wherein the composition is in the form of a gel, cream, ointment, or a liquid.21. The composition of embodiment 1-20, wherein the first and second components are combined into one dosage form.22. The composition of embodiment 1-20, wherein the first and second components are in separate dosage forms.23. The composition of embodiment 1-22, wherein the first component is in a form suitable for providing a daily substituted or unsubstituted dimdolylmethane dosage of about 10 mg to about 750 mg.24. The composition of embodiment 1-23, wherein the first component is in a. form suitable for providing a daily substituted or unsubstituted diindolyhnethane dosage of about 150 mg to about 650 mg, about 250 mg to about 550 mg, about300 mg to about 500 mg, about 400 mg to about 450 mg, or about 500 mg.25. The composition of embodiment 1 -24, wherein the first component is present in a dose which is a fraction of the daily dose, preferably a half of the daily dose, or a quarter of the daily dose.26. The composition of embodiment 1-25, wherein the composition is essentially free of any additional therapeutic agent.27. Use of the composition of embodiments 1 -26, for treating a skin condition.28. The use of embodiment 27, wherein the skin condition is an inflammatory skin condition.NON-LIMITING EXAMPLES

[0083] The following specific, non-limiting examples are to be construed as merely illustrative, and do not limit the present disclosure of the scope of the disclosure. Without further elaboration, it is believed that one skilled in the art can, based on the description herein, utilize the present disclosure to its fullest extent.Example 1: Oral DIM Compositions Containing Probiotic AgentManufacture of composition

[0084] A batch of DIM formulation containing an additional therapeutic agent was manufactured. The details of manufacture and formulation are set out below'.Formulation1 capsule contains:75 mg BR-DIM (BioResponse LLC)0.01-500mg of an additional therapeutic agent175 mg tri -calcium phosphate20 mg microcrystalline cellulose6 10 mg fumed, silicon dioxide (or a fine particle precipitated silica)6 mg magnesium stearateBlendingmethod for formulation

[0085] Mixing was carried out in a sequential process according to the following protocol:1. The BR-DIM was sieved through a fine sieve and then mixed with the magnesium stearate, half of the trical ciuni phosphate and the silicon dioxide for 15 mins in aV mixer. The mixed powder w-as transferred into a food grade bag.2. Separately, the additional therapeutic agent were mixed with the second half of the tricalcium phosphate and. the microcrystalline cellulose for 15 mins in a V mixer.3. The mixed powders obtained from 1 and 2 were blended together for 20 mins in aV mixer, then sieved, and mixed further for 1 hour and 45 minutes in a V mixer.4. The blended powder was transferred to a suitable opaque and air-tight container to ensure minimal contact with light and oxygen.5. A sample was taken from the top and bottom of the batch to test for uniformity of mixing.Encapsulation.

[0086] The DIM referred to above in some embodiments is encapsulated. The BR-DIM is spray dried into particles of approximately 8 to 12 microns in diameter. BR-DIM is known to be light sensitive. Therefore, opaque capsules are preferred, typically, size “0” capsules. Light resistant containers are preferred.Example 2 - Topical Gel DIM Compositions Containing Probiotic Agent

[0087] In this non-limiting example, the topical DIM composition is formulated as an emulsion gel, in which the oil phase contains the DIM compounds and the aquesous phase comprises a hydrogel. Below is a list of the ingredients for the emulsion gel formulation.[00881 Water - aquesous carrier

[0089] Glycerin - carrier

[0090] DIM - active agent (0.1-10 wt%)

[0091] Probiotic agent (0.1-10 wt%)

[0092] Dicaprylyl carbonate ester - emollient

[0093] Ammonium aciyloyldimethyltaurate / VP copolymer - rheology modifier

[0094] Carbomer - rheology modifier

[0095] Pol yglycery 1-10 laurate - emulsifier

[0096] Levulinic acid - skin conditioner and. preservative

[0097] Sodium anisate - preservative

[0098] Sodium levulinate - skin conditioner and preservative

[0099] Sodium hydroxide - pH adjusting agent

[0100] Tocopherol - moisturizer / antioxidant

[0101] To make the formulation, various ingredients were mixed together until a uniform emulsion gel is obtained.Example 3 - Topical Cream DIM Compositions Containing Probiotic Agent

[0102] In this non-limiting example, the topical DIM composition is formulated as an emulsion cream, in winch the oil phase contains the DIM compounds and the aqueous phase comprises a base cream. Below is a list of the ingredients for the emulsion cream formulation.

[0103] Water - aquesous carrier

[0104] Prunus amygdalus dulcis (sweet almond) oil

[0105] DIM - active agent (0.1-10 wt%)

[0106] Probiotic agent (0.1-10 wt%)

[0107] Dicaprylyl carbonate ester - emollient

[0108] Glvcerin - carrier

[0109] Stearyd alcohol fatty alcohol

[0110] Polyglyceryl-3 stearate - emulsifier

[0111] Potassium cetyl phosphate - emulsifier

[0112] Glyceryl stearate citrate - emulsifier

[0113] Xanthan gum - rheology7modifier / gum

[0114] Levulinic acid - skin conditioner and preservative

[0115] Ammonium acryloyldimethyltaurate / VP copolymer rheology7modifier - rheology modifier

[0116] Sodium anisate - preservative

[0117] Sodium levulinate - skin conditioner and preservative

[0118] Melianthus annuus (sunflower) seed oil

[0119] Sodium hydroxide - pH adjusting agent

[0120] Rosmarinus officinalis (rosemary') leaf extract

[0121] Tocopherol - moisturizer / antioxidant

[0122] To make the formulation, various ingredients were mixed together until a uniform emulsion cream is obtained.Example 4 - Clinical study of the effectiveness of oral DIM and a probiotic in patients with moderate to severe acne vulgarasIntroductionAn oral formulation of Diindolylmethane (DIM) + Vitamin A has been marketed as, or included in food supplements, in the UK and other countries since 2012, under the trade name of SKIN ACCUMAX™. It is currently sold with the claim to support ‘problem skin', which includes conditions such as acne. Studies conducted in Australia and USA have shown Skin Accumax™ (oral and sub-lingual respectively) to be effective at decreasing inflammation and improving acne. Case reports submitted by therapists over the years have supported this, showing benefit in acne and other inflammatory' skin conditions.An oral probiotic formulation, marketed under the tradename SKIN CLEAR BIOME™ was developed and is marketed specifically for problem skin. Case reports submitted by therapists over the years have also supported the benefit of this probiotic in the management of acne. We postulated, that DIM (without vitamin A) taken together with a. probiotic such as Skin Clear Biome™ would be more effective in treating acne than DIM (without vitamin A) alone or Skin Clear Biome™ alone.ObjectiveThis exploratory pilot study aimed to assess the effectiveness DIM taken together with a probiotic in patients with moderate to severe acne, versus the use of DIM alone or a probiotic such as Skin Clear Biome™ alone.Method3.1 Participants, procedure and product10 men and women, aged 18 to 41 years (average 25 yrs), with moderate to severe facial acne vulgaris (> 20 inflammatory' lesions / papules on the face, excluding the nose), were recruited for this trial. Participants were healthy, from a range of ethnic groups.Participants were excluded if they had used oral retinoids in the 3 months prior to screening, topical retinoids in the one month prior to screening, or were pregnant or lactating. If on hormonal contraception the prescription needed to remain the same over the 3 months prior to screening and for the duration of the trial. The use of nicotine was prohibited in the month before screening, and the duration of the trial.For 2 weeks prior to the onset of testing and for the duration of the trial, participants were instructed not to use prescribed medication, topical medication and / or self-tanning products, have any facial procedures / facials / chemical peels / dermabrasion, avoid excessive UV light exposure, take food supplements except for medically prescribed supplements such as iron or vitamin Bl 2, or drink more than 14 units of alcohol per week. They were also required to not make any changes to their skincare routine. The day before testing they were allowed to follow their normal skincare routine, ho wever on the day of testing, they were instructed to only wash their face / neck or affected areas with water and not wear any cosmetics. All subjects provided informed consent before participation.Participants were divided into 3 groups: group 1 (DIM+SCB) took 2 capsules of oral DIM twice daily (total daily dose of 75mg DIM) and I capsule of Skin Clear Biome™ once daily; group 2 (DIM only) took 2 capsules of oral DIM twice daily; and group 3 (SUB only) took 1 capsule of Skin Clear Biome™ once daily. One capsule of Skin Clear Biome™ contains 5 billion CPUs of Lactobacillus paracasei Lafti® L26 AF, Bifidobacterium bifidum Rosell®-71 (R0071 ), Lactobacillus helveticus Lafti® LI 0 ND and Saccharomyces boulardii, and Zinc 2mg. 3.2 AssessmentThis study was conducted o ver 16 weeks, with assessments at baseline (week 0) and weeks 4, 8, 12 and 16. Assessment consisted of physiological measurements, imaging, clinical assessments and self-assessment questionnaires.3.2.1 Physiological measurementsThe Courage & Khazaka™ equipment was used to measure properties of the skin. Courage & Khazaka™ are worldwide leaders in the production of dermatology testing devices.Direct measurement of skin using Courage & Khazaka™ probes: o Hydration in the stratum comeum is assessed with a Comeometer® which measures the capacitance of the di-eiectric properties of the stratum comeum. Hydration is measured in arbitrary units (AU) between 0 and 120, with very dry being < 30, dry being 30 - 40 and normal > 40. o Transepidermal water loss (TEWL) is assessed with a Tewameter® which uses open-chamber evaporimeters to measure TEWL. This together with the electrical method used by the Comeometer are the preferred techniques for measuring the water balance in the stratum comeum. TEWL is measured in g / li / 'm2, with values > 20 defined as increased.o Skin elasticity is assessed with a. Cutometer®, winch uses suction methods to deform skin and a non-contact optical measurement system to measure the deformity and recoil, which are used to calculate the elastic and viscoelastic properties of skin. There are several assessment parameters. We used R2 which measures the proportion between the amplitude after suction (Uf) and the ability to recover to the initial position (Ua). This is expressed as Ua / Uf (relaxation / suction) and is reported as a %. The higher the % the better. o Melanin and erythema (haemoglobin) are assessed with a Mexameter®, which emits 3 specific wavelengths of light and measures the reflectance of skin (light reflected back). Erythema is grouped into: none < 170, minimal 170 - 330, diffuse redness 330 - 450, high 450 - 750 and extreme > 570. Melanin varies per skin type. Both erythema and melanin are measured with arbitrary units (AU), between 0 -- 999. o Sebum levels are assessed with the Sebumeter®, which uses grease spot photometry. Sebum is measured in μg / cm2. On the forehead and t-zone, a value of < 100 is defined as dry / less sebum, 100 - 180 as normal, and > 180 as oily.3.2.2 Imaging<s High-resolution imaging of the skin surface with the Courage and Khazalca™ Visioscan®, a unique high resolution, UVA light video camera. This allows assessment of wrinkle depth, skin smoothness and skin roughness.Imaging of the full face using the E vends equipment, This uses cross-polarized and UV lighting to record and measure surface and subsurface skin erythema and pigmentation.3.2.3 Clinical assessments of acneThe Clinical Erythema Index (CEI)The CEI is a quantitative method for assessing the degree and extent of erythema which is one of the features of moderate and severe acne. The outcome is measured in each of four quadrants of the face. For each of the quadrants, a. score is given on an ordinal 5-point scale, with values ranging from 0 (none) to 4 (very severe). The total CEI score is a. sum of the four quadrants, giving a total score from 0 to 16.The Comprehensive Acne Severity Scale (CASS)The CASS is used to assess the severity of acne present on the face and / or upper back and chest.It consists of an ordinal 6-point scale, with values ranging from 0 (clear) to 5 (very severe).The Global Score for Inflammatory and Non-Inflammatosj LesionsAlthough various investigators have proposed acne severity grading schemata based upon counts and position on various quadrants of the body, none have been thoroughly validated. In this trial the number of non-inflammatory (NI) and inflammatory (I) lesions was counted, on the face (divided into 5 areas), the chest and the upper back. This approach has been accepted by the FDA in registration studies for Adapalene Cream 0.1 & 0.5% and Dapsone Gel 5%.& The Cardiff Acne Disability Index (CADI)This is a short, five-item questionnaire derived from the longer Acne Disability Index that is designed for use in adolescents and young adults to assess acne-related quality of life impairment. It is self-explanatory and can be simply handed to the patient who is asked to complete it without the need for detailed explanation. It is usually completed in one minute. A review by Abdelrazik et al., concluded that the questionnaire is reliable for measuring the impact of acne on the subject, but that no studies to date have yet calculated the smallest CADI score change required for the subject to have experienced a positive benefit from treatment. The CADI score is calculated by summing the score of each question resulting in a possible maximum of 15 and a minimum of 0. The higher the score, the more the quality of life is impaired,3,2.4 Self-assessment questionnaires s A self-assessment questionnaire was administered after initial application of the product, and at the end of weeks 1, 2, 4, 8, 12 and 16.3.3 Statistical analysis3.3.1 Normality Testing lire normality of the data sets was evaluated using the Data Analysis Toolpak in Microsoft Excel to guide the selection of appropriate statistical methods. Levene’s test showed that all data sets followed a. normal distribution, enabling the use of one-way ANOVA for comparing independent groups.3.3.2 ANOVA and t-testsTo compare the skin parameters, including stratum corneum hydration, elasticity, erythema, melanin, TEWL and sebum production one-way ANOVA was applied to analyse differences between each group over different time points (weeks 0, 4, 8, 12, and 16). Additionally, independent t-tests were applied to assess differences between each time point within each group.Individual subject analyses were also conducted to evaluate individual variations over the 16-week study period. One-way ANOVA was applied to assess differences within eachparticipant across different time points (weeks 0, 4, 8, 12, and 16), and independent t-tests were performed to compare changes between consecutive time intervals.Results4.1 DemographicsA total of 10 participants were included in the trial: 4 in the DIM+SCB group, 3 in the DIM only group and 3 in the SCB only group. One of the participants in the DIM+SCB group had only completed 12 weeks at the time of data analysis. The remainder of the participants completed the full 16 weeks. Table 1 show's the demographics of participants in the trial.Table 1. Demographics of the DIM+SCB, DIM only and SCB only groupsData between the 3 groups has been compared, however as the sample size of each group was small, the results of each individual are also presented.4,2 Physiological measurements4.2.1 BaselineAverage TEWL was slightly high in the DIM+SCB group (24.41 g / h / m2), while normal in the DIM only and SCB only groups (17.67 and 18.64 g / h / m2respectively); average hydration was in the normal range in all groups (56.39, 61.38 and 49.84 AU in the DIM+SCB, DIM only and SCB only groups respectively) and average erythema levels showed diffuse, approaching high ery thema in all groups (419.07, 407.97 and 430.06 AU in the DIM+SCB, DIM only and SCB only groups respectively). These changes are in keeping with the pathophysiology' of acne vulgaris, with erythema indicating inflammation and possible disruption of the skin barrier. Values are shows in table 2.4,2,2. During the trial - group resultsIn the DIM only group there was an increase in TEWL from 17.67 to 21.02 g / h / m2(p=0.05) from week 0 to 16, taking it from normal to just into the abnormal range. No other significant changes were seen. The lack of significance is likely a result of the small sample sizes. Results for sebum were not included due to several missing data, points as a result of equipment failure. Values are shown in table 2.Table 2. Measurement of skin parameters in the DIM+SCB group, DIM only group and SCB only group at weeks 0 (baseline), 4, 8, 12 and 164.2.3. During the trial - individual resultsDIM+SCB groupMeasurements of skin parameters are shown in table 3. Significant changes were seen in participants 3 and 4.Participant 3: baseline TEWL was high (39.35 g / h / m2). This decreased significantly over the 16 week trial with a 45% decrease from week 0 to week 8 (p=0.009), 38% decrease from week 0 to week 12 (p=0.015) and 43% decrease from week 0 to week 16 (p=0.005). There was also a significant increase in hydration over the trial with a. 39% increase from w'eek 0 to week 4 (p=0.007), 40% increase from week 0 to week 12 (p=0.008) and 52% increase from week 0 to week 16 (p=0.002).Participant 4: baseline erythema was high (508.67 AU). This decreased significantly over the 16 week trial with a 17% decrease from week 0 to week 4 (p=0.006), 14% decrease fromweek 0 to week 8 (p::::0.022) and 17% decrease to week 0 to week 12 (p::::0.002). The participant hadn’t yet had his 16 week appointment at the time of data analysis.Table 3. Measurement of skin parameters of each individual in the DIM+SCB group at weeks 0 (baseline), 4, 8, 12 and 16DIM only groupMeasurements of skin parameters are shown in table 4. Significant changes were seen in participant 3 only. Baseline erythema was high (493.80 AU). This decreased by 28% from week 0 to week 12 (p=0,012), however increased by week 16 so that there were no significant differences between weeks 0 and 16. There were no other significant changes.Table 4. Measurement of skin parameters of each individual in the DIM only group at weeks 0 (baseline), 4, 8, 12 and 16Week 0 Week 4 Week s Week 12 Week 16SOI only groupMeasurements of skin parameters are shown in table 5. Significant changes were seen in participant 1 only. Baseline TEWL was high (27.38 g / h / m2). This increased by 84% from week 0 to week 4 (p<0.0001) and by 35% from week 0 to week 16 (p:=:0.03). There were no other significant changes.Table 5. Measurement of skin parameters of each individual in the SCB only group at weeks 0 (baseline), 4, 8, 12 and 164.3 Clinical assessmentsThe CADI score in the DIM+SCB group improved by 6 points from week 0 to week 12 (p=0.008) and. by 7 points from week 0 to week 16 (p=0.015). No other significant changes were seen. Clinical assessment scores are shown in table 6.Table 6. Clinical assessment scores at weeks 0 (baseline), 4, 8, 12 and 16 in participants with acne vulgaris in in the DIM+SCB group, DIM only group and. SCB only group4.4 Self-assessments questionnairesTable 7 shows the percentage of participants who AGREED to self-assessment statements at weeks 4, 8 and 12. AH 4 participants in the DIM+SCB group agreed that continuous use of the products led to improvements in pigmentation at weeks 4, 8 and 12; improvements in acne, redness and hydration, and healthier looking skin at weeks 8 and 12; and improvements in blemishes, skin texture and reduced acne scarring at week 12.In the DIM only group all 3 participants reported improvements in acne, pigmentation and redness, and healthier looking skin at weeks 8 and 12; and improvements in blemishes, skin texture and scarring at week 12. Only 2 out of the 3 reported improved pigmentation at week 0, and improved hydration at weeks 8 and 12.In the SCB only group all 3 participants reported improved hydration and healthier looking skin at week 12. Only 2 out of the 3 reported improved pigmentation at weeks 4, 8 and. 12; improved acne, pigmentation, and redness at weeks 8 and 12; and only 1 of the 3 reported improved blemishes at week 12. The data is shown in table 7.Table 8 show's the percentage of participants who STRONGLY AGREED to self- assessment statements at week 16. All 4 participants in the DIM+SCB group strongly agreed that continuous use of the products decreased acne, pigmentation, redness, blemishes and acne scarring; improved hydration and skin texture and that their skin looked healthier. They also said that the products calmed and soothed, their skin and improved acne more than any other product tned.In the DIM only group, all 3 participants reported decreased redness, blemishes and acne scarring, and improved hydration. 2 out of 3 reported decreased, acne, improved skin texture andhealthier looking skin; and only 1 reported decreased pigmentation. AH 3 participants also said that the product calmed, and soothed their skin and improved acne more than any other product tried.In the SCB only group, 2 of the 3 participants reported decreased acne, redness, blemishes and acne scarring, and improved hydration, skin texture and healthier looking skin; and only 1 reported decreased pigmentation. 2 participants said that the product improved acne more than any other product tried and only 1 said that the product calmed and soothed their skin. Table 7. Participants in the DIM+SCB group, DIM only group and SCB only group who AGREED to the following statements after continuous use of the productsTable 8. Participants in the DIM+SCB group, DIM only group and SCB only group who STRONGLY AGREED to the following statemen ts after continuous use of the products at week 1DiscussionAt the start of the trial, participants of all 3 groups had diffuse approaching high erythema, indicating inflammation in keeping with the pathophysiology' of acne.Comparing the 3 groups, there was a slight increase in TEWL in the DIM only group. While this would suggest worsening of the skin barrier, scores were still very close to normal. No other significant changes were seen, likely because of the small sample sizes.Only the DIM+SCB group had a significant change in one of the assessment scores, with an improved CADI score over the trial, indicating improved quality of life.As the sample size of each group was small, the results of each individual were also analysed. Significant changes were seen in 2 of the participants in the DIM+SCB group. One participant had an improvement in TEWL and hydration indicating improved skin barrier function, with images showing decreased spots and redness. The second participant had an improvement in erythema indicating reduction in inflammation, which was also visible in the images.Significant changes were seen in only 1 participant in the DIM only group who had a reduction in ery thema, which was also seen in the images; and in only 1 participant in the SCB only group who had an increase in TEWL indicating potentially increased disruption of the skin barrier.Visual improvement was seen in the photographs and Eventis redness images in 3 of the participants in the DIM + SCB group, 2 in DIM only group and 1 in SCB only group.While all 3 groups had positive responses to the self-assessment questionnaires, the responses from the DIM+SCB were more consistently positive than the DIM only group which was more positive than the SCB only group.At 16 weeks, all 4 participants in the DIM+SCB group strongly agreed that the product improved acne, pigmentation, redness, blemishes, scarring, hydration and skin texture, and that their skin looked healthier. They also strongly agreed that the products calmed and soothed their skin and improved acne more than any other product tried. There were fewer positive responses m the DIM only group, and even fewer in the SCB only group.Safety outcomesNo adverse events were reported.ConclusionWhile improvements were seen in all 3 groups, the use of oral DIM together with Skin Clear Biome1Mled to more consistently positive results, despite the small sample sizes.

[0123] While specific embodiments described herein have been shown and described in detail to illustrate the application of the principles of the disclosure, it will be understood that the disclosure may be embodied otherwise without departing from such principles. Numerous variations, changes, and substitutions will be understood by those skilled in the art without departing from the disclosure. It should be understood, that various alternatives to the specific embodiments of the disclosure described herein may be employed in practicing the disclosure. It is intended that the claims define the scope of the disclos ure, and that methods and str uctures within the scope of these claims and their equivalents be covered thereby.

Claims

CLAIMSWhat is claimed as;1 . A composition for use in Treating a skin disease or condition, the composi tion comprising: a first component comprising a substituted or unsubstituted diindolylmethane: and a second component comprising at least one probiotic agent, wherein the composition is essentially free of substituted or unsubstituted retinoid compounds, wherein the composition is essentially free of CYP450 enzyme modulators, and wherein the composition is essentially free of anti-protozoal agents2. The composition of claim 1, wherein the composition further comprises a further component selected from one or more of, a substituted or unsubstituted azelaic acid, compound, an oral contraceptive compound, sulphur, a sulphur-containing compound, a substituted or unsubstituted salicylic acid compound, a substituted or unsubstituted resorcinol compound, a peroxide (optionally benzoyl peroxide), a plant product, a mineral, a vitamin, and a neutraceutical product.

3. The composition of claim I , wherein the first component comprises a diindolylmethane of Formula 1:wherein the R groups are independently selected from hydrogen atoms and Ci-Co hydrocarbon substituents; and wherein the indolyl groups are independently selected from indole-3-yl and mdole-2-yl groups: and wherein the indolyl groups are unsubstituted, or are substituted with one or more C1-C& hydrocarbon substituents.

4. The composition of claim 1, wherein the first component comprises a substituted or unsubstituted 3,3’diindolylmethane, of Formula 2. or a substituted or unsubstituted 2,2’diindolylmethane, of Formula 3:wherein the R groups are independently selected from hydrogen atoms, and C1-C6hydrocarbon substituents.

5. The composition of claim 1 , wherein the first component comprises a.3.3'diindolylmethane, of Formula 4 or a 2,2‘diindolylmethane, of Formula 5:

6. The composition of claim 1, wherein the first component comprises 3 , 3 ’ dii n d oly 1 in eth ane .

7. The composition of claim 1. wherein the first component is in a form suitable for providing a daily substituted or unsubstituted diindolylmethane dosage of up to 1500 mg or less.

8. The composition of claim 1, wherein the first component is in a form suitable for providing a daily substituted or unsubstituted diindolylmethane dosage of about 10 mg to about 750 mg.

9. The composition of claim 1 , wherein the substituted or unsubstituted diindolylmethane is adapted for increased bioavaiiability.

10. The composition of claim 9. wherein the substituted or unsubstituted diindolylmethane is microencapsulated.1 1. The composition of claim 10, wherein the first component comprises one or more lipophilic compounds selected from vitamin E and phosphatidylcholine.

12. The composition of claim 10, wherein the substituted or unsubstituted diindolylmethane is BR-DIM.

13. The composition of claim I , wherein the composition is formulated for oral administration.

14. The composition according to claim 13, wherein the composition is in the form of a tablet or capsule.

15. The composition of claim 1 , wherein the first and second components are combined into one dosage form.

16. The composition of claim 1, wherein the first and second components are in separate dosage forms.

17. The composition of claim 1, wherein the first component is in a form suitable for providing a daily substituted or unsubstituted diindolylmethane dosage of about 10 mg to about 750 mg.

18. The composition of claim 1. wherein the first component is in a form suitable for providing a daily substituted or unsubstituted diindolylmethane dosage of about 150 mg to about 650 mg, about 250 mg to about 550 mg, aboutSOO mg to about 500 mg. about 400 mg io about 450 mg. or about 500 mg.

19. The composition of claim 1, wherein the composition is essentially free of any additional therapeutic agent.

20. Use of the composition of claims 1, for treating a skin condition, optionally an inflammatory skin condition.

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