WO2024047527A1 - Methods and compositions for preventing and treating chemotherapy-induced peripheral neuropathy - Google Patents

Abstract

Disclosed herein are compositions, kits, and methods for preventing, ameliorating, or treating chemotherapy-induced peripheral neuropathy (CIPN) and/or reducing the severity of one or more risk factors, signs, or symptoms associated with CIPN. In particular, the technology of the present disclosure relates to methods for administering an effective amount of a composition comprising one or more strains of an operational group Bacillus amyloliquefaciens bacteria identified as ART12 and/or ART24 to a subject suffering from or at risk for CIPN.

Description

METHODS AND COMPOSITIONS FOR PREVENTING AND TREATING

CHEMOTHERAPY-INDUCED PERIPHERAL NEUROPATHY

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of and priority to U.S. Provisional Application No. 63/403,075, filed September 1, 2022, the contents of which are incorporated by reference in their entirety for any and all purposes.

TECHNICAL FIELD

[0002] The present technology relates to methods, compositions, and kits for preventing, ameliorating, or treating chemotherapy-induced peripheral neuropathy (CIPN) and/or reducing the severity of one or more risk factors, signs, or symptoms associated with CIPN. In particular, the present technology relates to administering an effective amount of a composition comprising one or more strains of an operational group Bacillus amyloliquefaciens bacteria, identified as ART 12 and ART24, to a subject suffering from or at risk for CIPN.

BACKGROUND

[0003] The following description is provided to assist the understanding of the reader. None of the information provided or references cited is admitted to be prior art to the compositions and methods disclosed herein.

[0004] Chemotherapy causes neuroinflammation and neurodegeneration leading to neuropathy. This is a debilitating problem for many cancer patients and is a limiting factor for chemotherapeutic treatment regimen. There is no standard of care for prevention of chemotherapy-induced peripheral neuropathy (CIPN). Accordingly, there is a need for additional therapies for preventing, treating, or reducing the risk of developing CIPN. SUMMARY

[0005] In one aspect, the disclosure of the present technology provides a method for treating or preventing chemotherapy-induced peripheral neuropathy (CIPN) in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising bacterial strain ART12 (NCIMB Accession No. 43087), ART24 (NCIMB Accession No. 43088), or a combination thereof In some embodiments, the bacterial strain is lyophilized. In some embodiments, the bacterial strain is in the form of a spore. In some embodiments, the bacterial strain is in vegetative form. In some embodiments, the bacterial strain is a mixture of the strain in the form of a spore and in the vegetative form. In some embodiments, the bacterial strain is formulated as a dietary supplement. In some embodiments, the bacterial strain is formulated as an edible product. In some embodiments, the edible product further comprises a carrier, vehicle, or excipient. In some embodiments, the edible product further comprises a fermented food product, soybean, mushroom, mung bean, locus bean, rice, or extracts thereof. In some embodiments, the soybean is fermented soybean or fermented soybean paste. In some embodiments, the fermented soybean or fermented soybean paste is Cheonggukjang, Douchi, Hawaijar, Bekang, Peruyaan, Tungrymbai, Eoyukjang, Kinema, Aakhone, Miso, Natto, or Thua-nao. In some embodiments, the CIPN comprises one or more of myalgias; paresthesia; glove-and-stocking sensory neuropathy, hyperalgia; allodynia, sharp, stabbing pain; burning or shock-like sensations; loss of sensation or complete numbness; trouble with small motor skills; gripping problems; clumsiness; weakness, and balance problems. In some embodiments, treatment or prevention comprises reducing allodynia. In some embodiments, the pharmaceutical composition is administered orally. In some embodiments, the method further comprises separately, sequentially, or simultaneously administering one or more agents selected from the group consisting of a steroid, lidocaine, capsaicin, an anti-seizure medication, an antidepressants, an analgesic, and any combination thereof. In some embodiments, the CIPN is associated with paclitaxel. In some embodiments, the CIPN is associated with cisplatin. [0006] In one aspect, the disclosure of the present technology provides a use of a composition in the preparation of a medicament for treating or preventing chemotherapy- induced peripheral neuropathy (CIPN) in a subject in need thereof, wherein the composition comprises bacterial strain ART 12 (NCIMB Accession No. 43087), ART24 (NCIMB Accession No. 43088), or a combination thereof. In some embodiments, the bacterial strain is lyophilized. In some embodiments, the bacterial strain is in the form of a spore. In some embodiments, the bacterial strain is in vegetative form. In some embodiments, the bacterial strain is a mixture of the strain in the form of a spore and in the vegetative form. In some embodiments, the bacterial strain is formulated as a dietary supplement. In some embodiments, the bacterial strain is formulated as an edible product. In some embodiments, the edible product further comprises a carrier, vehicle, or excipient. In some embodiments, the edible product further comprises a fermented food product, soybean, mushroom, mung bean, locus bean, rice, or extracts thereof. In some embodiments, the soybean is fermented soybean or fermented soybean paste. In some embodiments, the fermented soybean or fermented soybean paste is Cheonggukjang, Douchi, Hawaijar, Bekang, Peruyaan, Tungrymbai, Eoyukjang, Kinema, Aakhone, Miso, Natto, or Thua-nao. In some embodiments, the CIPN comprises one or more of myalgias; paresthesia; glove-and- stocking sensory neuropathy; hyperalgia; allodynia; sharp, stabbing pain; burning or shocklike sensations; loss of sensation or complete numbness; trouble with small motor skills; gripping problems; clumsiness; weakness; and balance problems. In some embodiments, treatment or prevention comprises reducing allodynia. In some embodiments, the pharmaceutical composition is formulated for oral administration. In some embodiments, the method further comprises separately, sequentially, or simultaneously administering one or more agents selected from the group consisting of a steroid, lidocaine, capsaicin, an antiseizure medication, an antidepressants, an analgesic, and any combination thereof. In some embodiments, the CIPN is associated with paclitaxel. In some embodiments, the CIPN is associated with cisplatin.

[0007] In one aspect, the disclosure of the present technology provides a composition used for treating or preventing chemotherapy-induced peripheral neuropathy (CIPN) in a subject in need thereof, wherein the composition comprises bacterial strain ART 12 (NCIMB Accession No. 43087), ART24 (NCIMB Accession No. 43088), or a combination thereof. In some embodiments, the bacterial strain is lyophilized. In some embodiments, the bacterial strain is in the form of a spore. In some embodiments, the bacterial strain is in vegetative form. In some embodiments, the bacterial strain is a mixture of the strain in the form of a spore and in the vegetative form. In some embodiments, the bacterial strain is formulated as a dietary' supplement. In some embodiments, the bacterial strain is formulated as an edible product. In some embodiments, the edible product further comprises a carrier, vehicle, or excipient. In some embodiments, the edible product further comprises a fermented food product, soybean, mushroom, mung bean, locus bean, rice, or extracts thereof. In some embodiments, the soybean is fermented soybean or fermented soybean paste. In some embodiments, the fermented soybean or fermented soybean paste is Cheonggukjang, Douchi, Hawaijar, Bekang, Peruyaan, Tungrymbai, Eoyukjang, Kineraa, Aakhone, Miso, Natto, or Thua-nao. In some embodiments, the CIPN comprises one or more of myalgias; paresthesia; glove-and-stocking sensory neuropathy, hyperalgia; allodynia; sharp, stabbing pain; burning or shock-like sensations; loss of sensation or complete numbness; trouble with small motor skills; gripping problems; clumsiness; weakness; and balance problems. In some embodiments, the treatment or prevention comprises reducing allodynia. In some embodiments, the pharmaceutical composition is formulated for oral administration. In some embodiments, the composition for use further comprises separately, sequentially, or simultaneously administering one or more agents selected from the group consisting of a steroid, lidocaine, capsaicin, an anti-seizure medication, an antidepressants, an analgesic, and any combination thereof. In some embodiments, the CIPN is associated with paclitaxel. In some embodiments, the CIPN is associated with cisplatin.

[0008] In one aspect, the disclosure of the present technology provides a kit for use in treating or preventing CIPN comprising: (a) a composition comprising a bacterial strain selected from the group consisting of: ART12 (NCIMB Accession No. 43087) and ART24 (NCIMB Accession No. 43088); and (b) a package insert with instructions for treating or preventing CIPN in a subject in need thereof. In some embodiments, the kit of claim 52, further comprising a co-therapeutic selected from the group consisting of a steroid, lidocaine, capsaicin, an anti-seizure medication, an antidepressants, an analgesic, and any combination thereof.

BRIEF DESCRIPTION OF THE DRAWINGS

[0009] Figure 1A is a schematic depiction of Taxol induction and treatment.

[0010] Figure IB is a chart showing the mean group response to von Frey force (g) at, from left to right on the x-axis, Baseline, Day 8, and Day 14. Each group (Groups 1-6) is shown at each timepoint, from left to right along the x-axis as: Placebo/no Taxol (Group 1); Placebo (Group 2); ART24 (Group 3); Cryo (Group 4); ART12 (Group 5); and Gabapentin 100 mg/kg (Group 6). # p<0.05 vs. Placebo (Group 2) using Student’s T-test; ## p<0.01 vs. Placebo (Group 2) using Student’s T-test; **** p<0.0001 vs. Placebo (Group 2) using one- way ANOVA, followed by Dunnett’s post-hoc test.

[0011] Figure 2A is a schematic depiction of cisplatin induction and treatment.

[0012] Figure 2B is a chart, showing the mean group response to von Frey force (g) at, from left to right on the x-axis, Baseline, Day 15, and Day 18. Each group (Groups 1-5) is shown at each timepoint, from left to right along the x-axis as: Saline + PBS (Group 1); Cisplatin + PBS (Group 2); Cisplatin + ADS012 (Group 3); Cisplatin + ADS024 (Group 4); and Cisplatin + Gabapentin (Group 5)

. p<0.0001 vs. Cisplatin +PBS (Group 2) using a one-

way ANOVA test followed by Dunnett’s test; p<0.0001 vs. Saline +PBS (Group 1) using student’s T-test.

DETAILED DESCRIPTION

[0013] It is to be appreciated that certain aspects, modes, embodiments, variations and features of the present technology are described below in vari ous levels of detail in order to provide a substantial understanding of the present technology. The definitions of certain terms as used in this specification are provided below. Unless defined otherwise, all technical and scientific terms used herein generally have the same meaning as commonly understood by one of ordinary' skill in the art to which this present technology belongs. Definitions

[0014] The following terms are used herein, the definitions of which are provided for guidance.

[0015] As used herein, the singular forms “a,” “an,” and “the” designate both the singular and the plural, unless expressly stated to designate the singular only.

[0016] The term “about” and the use of ranges in general, whether or not qualified by the term about, means that the number comprehended is not limited to the exact number set forth herein, and is intended to refer to ranges substantially within the quoted range while not departing from the scope of the present technology. As used herein, “about” will be understood by persons of ordinary skill in the art and will vary to some extent on the context in which it is used. If there are uses of the term which are not clear to persons of ordinary skill in the art given the context in which it is used, “about” will mean up to plus or minus 10% of the particular term.

[0017] As used herein, “administration” of an agent, drug, bacterial strain(s) or spore thereof, or composition of the present technology to a subject includes any route of introducing or delivering to a subject a compound to perform its intended function. Administration can be carried out by any suitable route, including orally, intranasally, parenterally (intravenously, intramuscularly, intraperitoneally, or subcutaneously), topically, or by inhalation. In some embodiments, the compositions of the present technology are formulated for enteric administration. In some embodiments, the compositions are formulated for oral, sublingual, or rectal delivery. In some embodiments, the compositions are formulated for use as a probiotic. In some embodiments, the compositions are formulated for use as a live biotherapeutic. As used herein, administration includes self-administration and administration by another.

[0018] As used herein, “ART12” or “ADS012” refers to a bacterial strain, or spore thereof, having been deposited under NCIMB Accession No. 43087, or compositions comprising the strain. ART 12 is considered a member of an “operational group B. amyloliquefaciens that comprises the soil-borne B. amyloliquefaciens, and plant associated Bacillus siamensis and Bacillus velezensis.

[0019] As used herein, “ART24” or “ADS024” refers to a bacterial strain, or spore thereof, having been deposited under NCIMB Accession No. 43088, or compositions comprising the strain. ART24 is considered a member of an “operational group B. amyloliquefac lens” that comprises the soil-borne B. amyloliquefaciens, and plant associated Bacillus siamensis and Bacillus velezensis.

[0020] As used herein, the terms “effective amount,” or “therapeutically effective amount,” and “pharmaceutically effective amount” refer to a quantity sufficient to achieve a desired therapeutic and/or prophylactic effect, e.g., an amount which results in the prevention of a disease, condition, and/or symptom(s) thereof. In the context of therapeutic or prophylactic applications, the amount of a composition administered to the subject will depend on the type and severity of the disease and on the characteristics of the subject, such as general health, age, sex, body weight, and tolerance to the composition drugs. It will also depend on the degree, severity, and type of disease or condition. The skilled artisan will be able to determine appropriate dosages depending on these and other factors. In some embodiments, multiple doses are administered. Additionally or alternatively, in some embodiments, multiple therapeutic compositions or compounds (e.g., pharmaceutical compositions comprising the bacterial strain(s) alone or in combination with additional active agents, such as steroids, lidocaine, capsaicin, anti-seizure medications, antidepressants, or analgesics, such as narcotics or opiate drugs, are administered. In the methods described herein, compositions comprising the bacterial strain(s) of the present technology, or spores thereof, may be administered to a subject having one or more signs, symptoms, or risk factors of CIPN, including, but not limited to, myalgias; painful burning paresthesia; glove-and-stocking sensory neuropathy; hyperalgia; allodynia; paresthesia (tingling or pins-and-needles sensation); sharp, stabbing pain; burning or shock-like sensations; loss of sensation or complete numbness; trouble with small motor skills such as writing, texting, and buttoning; gripping problems (dropping items); clumsiness; weakness, especially in the hands and feet; and/or balance problems. For example, a “therapeutically effective amount” of the compositions of the present technology, includes levels at which the presence, frequency, or severity of one or more signs, symptoms, or risk factors of CIPN are, at a minimum, ameliorated. In some embodiments, a therapeutically effective amount reduces or ameliorates the physiological effects of CIPN, and/or the risk factors of CIPN, and/or the likelihood of developing CIPN. In some embodiments, a therapeutically effective amount is achieved by multiple administrations. In some embodiments, a therapeutically effective amount is achieved with a single administration.

[0021] As used herein, the terms “freeze-dried” or “freeze-drying” and “lyophilized” or “lyophilization” are used interchangeably and refer to a process that removes water from a product after it is frozen and placed under a vacuum and the products produced therefrom.

[0022] As used herein, “pharmaceutically acceptable carrier and/or diluent” or “pharmaceutically acceptable excipient” includes but is not limited to solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like. In some embodiments, the pharmaceutically acceptable carrier comprises a polysaccharide, locust bean gum, an anionic polysaccharide, a starch, a protein, sodium ascorbate, glutathione, trehalose, sucrose, or pectin. In some embodiments, the polysaccharide comprises a plant, animal, algal, or microbial polysaccharide. In some embodiments, the polysaccharide comprises guar gum, inulin, amylose, chitosan, chondroitin sulphate, an alginate, or dextran. In some embodiments, the starch comprises rice starch. The use of such media and agents for biologically active substances is well known in the art. Further details of excipients are provided below. Supplementary active ingredients, such as antimicrobials, for example antifungal agents, can also be incorporated into the compositions.

[0023] As used herein, “pharmaceutically acceptable excipient” refers to substances and compositions that do not produce an adverse, allergic, or other untoward reaction when administered to an animal or a human. As used herein, the term includes all inert, non- toxic, liquid or solid fillers, or diluents that do not react with the therapeutic substance of the present technology in an inappropriate negative manner, solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, preservatives and the like, for example liquid pharmaceutical carriers e.g., sterile water, saline, sugar solutions, Tris buffer, ethanol and/or certain oils.

[0024] As used herein, “probiotic” refers to bacteria comprising a component of the transient or endogenous flora of a subject administered to confer a beneficial prophylactic and/or therapeutic effect on the subject. Probiotics are generally known to be safe by those skilled in the art.

[0025] As used herein, “live biotherapeutic product(s)” refers to live microorganisms that are applicable to the prevention, treatment, or cure of a disease, condition, or disorder. Live biotherapeutic products can further comprise co-therapeutics, pharmaceutically acceptable excipients, or other useful compounds.

[0026] As used herein, “prevention,” “prevent,” or “preventing” of a disorder or condition refers to, in a statistical sample, reduction in the occurrence or recurrence of the disorder or condition in treated subjects/samples relative to an untreated controls, or refers delays the onset of one or more symptoms of the disorder or condition relative to the untreated controls.

[0027] As used herein “subject” and “patient” are used interchangeably. In some embodiments, the subject is an animal subject. In some embodiments, the animal subject is a mammal. In some embodiments, the mammalian subject is a human.

[0028] As used herein, the term “simultaneous” administration refers to the administration of at least two agents by the same route and at the same time or at substantially the same time.

[0029] As used herein, the term “separate” administration refers to an administration of at least two agents at the same time or at substantially the same time by different routes.

[0030] As used herein, the term “sequential” administration refers to administration of at least two agents at different times, the administration route being identical or different. More particularly, sequential use refers to the whole administration of one agent before administration of the other agent(s) commences. It is thus possible to administer one of the agents over several minutes, hours, or days before administering another.

[0031] A “synergistic therapeutic effect” refers to a greater-than-additive therapeutic effect which is produced by a combination of at least two therapeutic agents, and which exceeds that which would otherwise result from the individual administration of the agents. For example, use of the bacterial strain(s) of the present technology in conjunction with other agents for the treatment of CIPN may result in a greater than additive therapeutic effect. In some embodiments, the synergistic effect may permit the use of lower doses of the bacterial strain(s) of the present technology and/or other agents than would be required if each were used alone.

[0032] “Treating,” “treat,” “treated,” or “treatment” of a disease or disorder includes: (i) inhibiting the disease or disorder, i.e.. arresting its development; (ii) relieving the disease or disorder, i.e., causing its regression; (iii) slowing progression of the disorder, and/or (i v) inhibiting, relieving, or slowing progression of one or more symptoms of the disease or disorder. In some embodiments, treating a disease or disorder comprises ameliorating the disease or disorder or a symptom of the disease or disorder.

[0033] It is to be appreciated that the various modes of treatment or prevention of medical diseases and conditions as described are intended to mean “substantial,” which includes total but also less than total treatment or prevention, and wherein some biologically or medically relevant result is achieved.

II. Chemotherapy-Induced Peripheral Neuropathy (CIPN)

[0034] The ART12 and/or ART24 strains, or spores thereof, described herein are useful in methods for treating or preventing chemotherapy -induced peripheral neuropathy (CIPN). In some embodiments the ART 12 and/or ART24 strains, or spores thereof, may be administered to a subject following the onset of CIPN. Thus, the term “treatment” is used herein in its broadest sense and refers to use of an ART 12 and/or ART24 strain, or spores thereof, for a partial or complete cure of the CIPN, a reduction or amelioration of signs or symptoms, and/or a reduction of severity of signs or symptoms.

[0035] In some embodiments, the ART12 and/or ART24 strains, or spores thereof, may be administered to a subject before the onset of CIPN in order to prevent, protect against, and/or provide prophylaxis for neuropathy or hyperalgesia. It is also contemplated that the strains, or spores thereof, may be administered to a subject at risk of developing CIPN.

[0036] The term “peripheral neuropathy” refers generally to damage to nerves of the peripheral nervous system. .As used herein, the “peripheral neuropathy” encompasses motor, sensory, mixed sensorimotor, chronic, and acute neuropathy. As used herein the term encompasses mononeuropathy, multiple mononeuropathy, and polyneuropathy . When chemotherapeutic agents cause degeneration of peripheral sensory and motor nerves, it is referred to as “chemotherapy-induced peripheral neuropathy” or “CIPN.”

[0037] Chemotherapy -induced peripheral neuropathy can cause a variety of dose-limiting neuropathic conditions, including 1) myalgias, 2) painful burning paresthesis, 3) glove-and- stocking sensory neuropathy, and 4) hyperalgia and allodynia. Hyperalgia refers to hypersensitivity and pain caused by stimuli that is normally only mildly painful or irritating. Allodynia refers to hypersensitivity and pain caused by stimuli that is normally not painful or irritating.

[0038] The signs and symptoms of CIPN depend on the type of chemotherapy and which nerve fibers are affected. In addition to the neuropathic conditions described above, subjects with CIPN may experience: paresthesia (tingling or pins-and-needles sensation); sharp, stabbing pain; burning or shock-like sensations; loss of sensation or complete numbness; trouble with small motor skills such as writing, texting, and buttoning, gripping problems (dropping items); clumsiness; weakness, especially in the hands and feet; and/or balance problems. CIPN can last for weeks, months, or even years after the completion of chemotherapy. If it progresses, CIPN can cause more serious problems such as changes in heart rate and blood pressure, falling, breathing difficulties, paralysis, or organ failure. [0039] The following are non-limiting examples of chemotherapeutics that may cause CIPN: platinum drugs (e.g., cisplatin, carboplatin, oxaliplatin); taxanes (e.g., paclitaxel (Taxol®), docetaxel (Taxotere®, cabazitaxel (Jevtana®)); plant alkaloids (e.g., vinblastine, vincristine, vinorelbine, etoposide (VP-16)); immunomodulating drags (e.g., thalidomide (Thalomid®), lenalidomide (Revlimid®), pomalidomide (Pomalyst®)); and proteasome inhibitors (e.g., bortezomib (Velcade®), carfilzomib (Kyprolis®), and ixazomib (Ninlaro)). In some embodiments, the ART 12 and/or ART24 strains, or spores thereof, described herein are useful in methods for treating or preventing CIPN associated with the use of any one or more of the preceding chemotherapeutics. In some embodiments, the ART12 and/or ART24 strains, or spores thereof, described herein are useful in methods for treating or preventing CIPN associated with the use of cisplatin. In some embodiments, the ART12 and/or ART24 strains, or spores thereof, described herein are useful in methods for treating or preventing CIPN associated with the use of paclitaxel (Taxol®).

[0040] Diagnosis of CIPN is typically based on a subject’s medical hi story, clinical examination, and supporting lab tests, which include electromyography with nerve induction studies, skin biopsies to evaluate cutaneous nerve innervation, and nerve and muscle biopsies for histopathological evaluation.

[0041] Treatment of CIPN depends on discontinuati on or lowering the dose of the chemotherapeutic. Persistent neuropathic pain can be treated with steroids, lidocaine, capsaicin, anti-seizure medications, antidepressants, or analgesics, such as narcotics or opiate drugs. In severe conditions, patients may be referred to pain management clinics. Subjects suffering from severe balance problems may benefit from vestibular rehabilitation.

[0042] In some embodiments, the present technology provides methods and compositions for the treating or preventing CIPN, including reducing the severity of one or more risk factors, signs, or symptoms associated with CIPN. In some embodiments the compositions comprise strain ART 12 and/or ART24, or spores thereof. III. ART 12 and ART24

[0043] The technology of the present disclosure relates to the use of the ART12 strain and/or ART24 strain, or spores thereof, to treat or prevent CIPN. ART 12 refers to a bacterial strain, or spore thereof, having been deposited under NCIMB Accession No. 43087, or compositions comprising the strain. ART24 refers to a bacterial strain, or spore thereof, having been deposited under NCIMB Accession No. 43088, or compositions comprising the strain.

[0044] In some embodiments, the bacterial strain(s) of the present technology, or spores thereof, is used in methods and compositions for treating or preventing CIPN. In some embodiments, the bacterial strain(s) comprises a probiotic for preventing or controlling CIPN. In some embodiments, the bacterial strain(s) comprises a live biotherapeutic product for preventing or controlling CIPN. In some embodiments, compositions of the present technology comprise vegetative bacterial cells. In some embodiments, compositions of the present technology comprise bacterial spores. In some embodiments, compositions of the present technology comprise a combination of vegetative bacterial cells and bacterial spores. In some embodiments, the compositions of the present technology comprise ART12. In some embodiments, the compositions of the present technology comprise ART24. In some embodiments, the compositions of the present technology comprise ART12 and ART24. In some embodiments, the compositions of the present technology comprise an ART12 and/or ART24 bacterial strain that has been physically destructed or lysed. In some embodiments, the compositions of the present technology comprise an isolated fraction of lysed ART12 and/or ART24 bacteria.

[0045] In some embodiments, the bacterial strain(s) of the present technology, or spores thereof, is used in methods and edible product compositions for treating or preventing CIPN in a subject in need thereof. In some embodiments, the bacterial strain(s) comprises a dietary supplement for preventing or controlling CIPN in a subject in need thereof. IV. Therapeutic and Prophylactic Methods

[0046] The following discussion is presented by way of example only, and is not intended to be limiting.

[0047] One aspect of the present technology includes methods of treating or preventing CIPN in a subject diagnosed as having, suspected as having, or at risk of having CIPN. In therapeutic applications, compositions or medicaments comprising the ART 12 and/or ART24 bacterial strain, or spore thereof, are administered to a subject suspected of, or already suffering from CIPN (such as, e.g., subjects exhibiting one or more signs or symptoms of CIPN), in an amount sufficient to cure, or at least partially arrest, the signs or symptoms of the CIPN, including its complications and intermediate pathological phenotypes in development of the disease.

[0048] Subjects suffering from CIPN can be identified by any or a combination of diagnostic or prognostic assays known in the art. For example, typical symptoms of CIPN include, but are not limited to: myalgias; painful burning paresthesia; glove-and-stocking sensory neuropathy; hyperalgia; allodynia, paresthesia (tingling or pins-and-needles sensation); sharp, stabbing pain; burning or shock-like sensations; loss of sensation or complete numbness, trouble with small motor skills such as writing, texting, and buttoning; gripping problems (dropping items); clumsiness; weakness, especially in the hands and feet; and/or balance problems.

[0049] In some embodiments, subjects with CIPN treated with the bacterial strain(s) of the present technology, or spores thereof, will show amelioration or elimination of one or more of the following symptoms: myalgias; painful burning paresthesia; glove-and-stocking sensory neuropathy; hyperalgia, allodynia; paresthesia (tingling or pins-and-needles sensation); sharp, stabbing pain; burning or shock-like sensations; loss of sensation or complete numbness; trouble with small motor skills such as writing, texting, and buttoning, gripping problems (dropping items); clumsiness; weakness, especially in the hands and feet; and/or balance problems. [0050] In one aspect, the present technology provides a method for preventing or delaying the onset of CIPN or symptoms of CIPN in a subject at risk of having CIPN. In some embodiments, the bacterial strain(s) of the present technology is formulated as a probiotic useful as a food supplement and for re-establishing beneficial bacteria in the intestinal tract. In some embodiments, the bacterial strain(s) of the present technology is formulated as a live biotherapeutic product useful in pharmaceutical applications. In some embodiments, the bacterial strain(s) of the present technology i s formulated as a live biotherapeutic edible product useful in pharmaceutical applications.

V. Modes of Administration and Effective Dosages

[0051] Compositions of the present technology for use in preventing, ameliorating, or treating CIPN and/or reducing the severity of one or more risk factors, signs, or symptoms associated with CIPN include live probiotic ART12 and/or ART24 bacteria according to the present technology, provided in the form of vegetative cells and/or spores. In some embodiments, the bacterial strain is formulated as a live biotherapeutic product. In some embodiments, the bacterial strain is formulated as a probiotic. In some embodiments, the bacterial strain(s) is lyophilized. The compositions of the present technology are administered to the subject in effective amounts (i.e., amounts that have desired therapeutic effect). The dose and dosage regimen will depend upon the degree of disease, the characteristics of the ART12 and/or ART24 strain used, e.g., its therapeutic index, the subject, and the subject’s history. The effective amount may be determined during pre- clinical trials and clinical trials by methods familiar to physicians and clinicians.

[0052] Compositions of the present technology may be formulated for adding to food, or used directly as a food supplement. The formulation may further include other probiotic agents or nutrients for promoting spore germination and/or bacterial growth.

[0053] Edible products of the present technology may be formulated for adding to food, or used directly as a dietary supplement. In some embodiments, the edible product comprises a fermented food product, soybean, mushroom, mung bean, locus bean, or rice. In some embodiments, the soybean is fermented soybean or fermented soybean paste. The formulation may further include other probiotic agents or nutrients for promoting spore germination and/or bacterial growth.

[0054] Additional components of the compositions of the present technology may include a preservative selected from the group consisting of sucrose, sodium ascorbate, and glutathione. In some embodiments the preservative is a cryoprotectant selected from the group consisting of a nucleotide, a disaccharide, a polyol, and a polysaccharide. In some embodiments, the cryoprotectant is selected from the group consisting of inosine-5’- monophosphate (IMP), guanosine-5 ’-monophosphate (GMP), adenosine-5’ -monophosphate (AMP), uranosine-5’ -monophosphate (UMP), cytidine-5’-monophosphate (CMP), adenine, guanine, uracil, cytosine, guanosine, uridine, cytidine, hypoxanthine, xanthine, orotidine, thymidine, inosine, trehalose, maltose, lactose, sucrose, sorbitol, mannitol, dextrin, inulin, sodium ascorbate, glutathione, skim milk, and cryoprotectant 18.

[0055] The ART 12 and/or ART24 bacterial strain described herein can be incorporated into pharmaceutical compositions for administration, singly or in combination, and given to a subject for the treatment or prevention of a disorder described herein. Such compositions typically include the active agent and a pharmaceutically acceptable carrier. As used herein the term “pharmaceutically acceptable carrier” includes saline, solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like, compatible with pharmaceutical administration. Supplementary' active compounds can also be incorporated into the compositions. Carriers can be solid-based dry materials for formulations in powdered form, and can be liquid or gel-based materials for formulations in liquid or gel forms, which forms depend, in part, upon the routes or modes of administration. In some embodiments, the pharmaceutically acceptable carrier comprises a polysaccharide, locust bean gum, an anionic polysaccharide, a starch, a protein, sodium ascorbate, glutathione, trehalose, sucrose, or pectin. In some embodiments, the polysaccharide comprises a plant, animal, algal, or microbial polysaccharide. In some embodiments, the polysaccharide comprises guar gum, inulin, amylose, chitosan, chondroitin sulphate, an alginate, or dextran. In some embodiments, the starch comprises rice starch. [0056] Pharmaceutical compositions are typically formulated to be compatible with its intended route of administration. Examples of routes of administration include enteric (e.g, oral, sublingual, rectal) administration. A therapeutic composition can be formulated to be suitable for oral administration in a variety of ways, for example in a liquid, a powdered food supplement, a solid food, a packaged food, a wafer, tablets, troches, or capsules, e.g., gelatin capsules, and the like. In some embodiments, the therapeutic compositions of the present technology comprise lyophilized AR.T12 and/or ART24. In some embodiments, the lyophilized ART12 and/or ART24 is encapsulated. A therapeutic composition can be formulated to be suitable for rectal administration in a variety of ways, for example in a suppository, liquid enema, or foam. Other formulations will be readily apparent to one skilled in the art. pH can be adjusted with acids or bases, such as hydrochloric acid or sodium hydroxide.

[0057] Additionally or alternatively, for edible products, pharmaceutical compositions of the present technology may be formulated as dietary' supplements, nutraceutical compositions, food additives, food compositions, food compositions in bulk, food additives in bulk, medical foods, or foods for special dietary use, for enteric (e.g, oral, sublingual) administration.

[0058] Dosage, toxicity and therapeutic effi cacy of any therapeutic agent can be determined by standard pharmaceutical procedures in cell cultures or experimental animals. The data obtained from the cell culture assays and animal studies can be used in formulating a range of dosage for use in humans. The dosage of such compounds may be within a range of circulating concentrations that include the ED50 with little or no toxicity'. The dosage may vary within this range depending upon the dosage form employed and the route of administration utilized. For any compound used in the methods, the therapeutically effective dose can be estimated initially from cell culture assays. A dose can be for mulated in animal models to achieve a circulating plasma concentration range that includes the IC50 (i.e., the concentrati on of the test compound which achieves a half-maximal i nhibition of symptoms) as determined in cell culture. Such information can be used to determine useful doses in humans accurately. [0059] In some embodiments, the “edible products” such as whole soybeans and/or soybean pastes, are inoculated with a concentration of ART 12 and/or ART24 ranging from at least about 1 x 10² colony forming units (CFU)/mL to at least about 1 x 10¹² CFU/mL, or any value in between. For example, in some embodiments, the edible products are inoculated with at least about 1 x 10³ CFU/mL to at least about 1 x 10¹² CFU/mL ART12 and/or ART '24, at least about 1 x 10⁴ CFU/mL to at least about 1 x 10¹² CFU/mL ART 12 and/or ART24, at least about 1 x 10⁵ CFU/mL to at least about 1 x 10¹² CFU/mL ART12 and/or ART24, at least about 1 x 10⁶ CFU/mL to at least about 1 x 10¹² CFU/mL ART12 and/or ART24, at least about l x 10⁷ CFU/mL to at least about 1 x 10¹² CFU/mL ART 12 and/or ART24, at least about 1 x 10⁸ CFU/mL to at least about 1 x 10¹² CFU/mL ART 12 and/or ART24, at least about 1 x 10⁹ CFU/mL to at least about 1 x 10¹² CFU/mL ART12 and/or ART24, at least about 1 x 10¹⁰ CFU/mL to at least about 1 x 10¹² CFU/mL ART12 and/or ART24, at least about 1 x 10¹¹ CFU/mL to at least about 1 x 10¹² CFU/mL ART12 and/or ART24. In some embodiments, the edible products are inoculated with at least about 6 x 10⁸ CFU/mL ART12 and/or ART24. In some embodiments, the edible products are inoculated with at least about 6 x 10⁹ CFU/mL ART 12 and/or ART24. In some embodiments, the edible products are inoculated with at least about 1 x 10⁴ CFU/mL ART12 and/or ART24. In some embodiments, the edible products are inoculated with at least about 1 x 10⁴ CFU/mL ART 12 and/or ART24. In some embodiments, the edible products are inoculated with at least about 1 x 10⁶ CFU/mL ART12 and/or ART24. In some embodiments, the edible products are inoculated with at least about 1 x 10⁸ CFU/mL ART12 and/or ART24. In some embodiments, the edible products are inoculated with at least about 1 x 10⁹ CFU/mL ART12 and/or ART24. In some embodiments, the edible products are inoculated with at least about 1 x 10¹⁰ CFU/mL ART12 and/or ART24. In some embodiments, the edible products are inoculated with at least about 1 x 10¹¹ CFU/mL ART12 and/or ART24. In some embodiments, the edible products are inoculated with at least about 1 x 10¹² CFU/mL ART 12 and/or ART24.

[0060] In some embodiments, the compositions of the present technology are formulated at about 1 x 10⁸ col ony forming units (CFU) per ml to about 1x 10¹² CFU per ml of ART24 and/or ART12 bacterium (ie., vegetative cell) or bacterial spore. In some embodiments, the methods of the present technology involve the administration of about 1x10⁹to about 1x10¹² viable bacteria or spores per day. In some embodiments, the compositions of the present technology are delivered as lyophilized material or powder to be re-suspended for oral delivery or packaged into capsules. The lyophilized material and capsules may be coated for better enteric stability.

[0061] An exemplary treatment regimen entails administration once per day or once a week. In therapeutic applications, a relatively high dosage at relatively short, intervals is sometimes required until progression of the disease is reduced or terminated, or until the subject shows partial or complete amelioration of symptoms of disease. Thereafter, the subject can be administered a prophylactic regime. In some embodiments, compositions of the present technology are administered to a subject multiple times per day. In some embodiments, compositions of the present technology are administered to a subject once, twice, or three times per day or more for a certain period of time or until the subject is deemed cured of primary' disease, not to be at risk for recurrence of primary disease, or not to be at risk for the disease. In some embodiments, the composi tions of the present technology may be administered to the subject for the remainder of the subject’s life. In some embodiments, administration is paired with a shortened exposure to agents known in the art for the treatment of CIPN, such as steroids, lidocaine, capsaicin, anti-seizure medications, antidepressants, or analgesics, such as narcotics or opiate drags, followed by once, twice, or three times daily dosing or more for a certain period of time or until the patient is deemed cured of primary disease or not to be at risk for recurrence of the disease. In some embodiments, methods of prophylaxis comprise administration of compositions of the present technology once, twice, or three times daily or more for a certain period of time or until the patient, is deemed not to be at risk of developing the disease in the case of patients known to be at risk for CIPN.

[0062] The skilled artisan will appreciate that certain factors may influence the dosage and timing required to effectively treat, a subject, including but. not limited to, the severity of the disease or disorder, previous treatments, the general health and/or age of the subject, and other diseases present. Moreover, treatment of a subject with a therapeutically effective amount of the therapeutic compositions described herein can include a single treatment or a series of treatments.

VI. Combination Therapy with ART 12 and/or ART24

[0063] In some embodiments, the ART12 and/or ART24 strain of the present technology, or spores thereof, may be combined with one or more additional therapeutic regimens. In some embodiments, the additional therapeutic regimens are directed to the treatment or prevention of CIPN or symptoms associated with CIPN. In some embodiments, the additional therapeutic regimens comprise administration of one or more drugs, including, but not limited to, steroids, lidocaine, capsaicin, anti-seizure medications, antidepressants, and analgesics. In embodiments, the additional therapeutic regimens comprise nonpharmaceutical therapies, including, but not limited to, pain management regimens and/or vestibular rehabilitation.

[0064] In some embodiments, an additional therapeutic agent is administered to a subject in combination with the ART12 and/or ART24 strain of the present technology, or spores thereof, such that a synergistic therapeutic effect is produced. For example, administration of ART 12 and/or ART24 with one or more additional therapeutic agents for the prevention or treatment of CIPN will have greater than additive effects in the prevention or treatment of the disease and/or one or more of its signs or symptoms.

[0065] In any case, the multiple therapeutic agents may be administered in any order or even simultaneously. If simultaneously, the multiple therapeutic agents may be provided in a single, unified form, or in multiple forms (by way of example only, either as a single pill or as two separate pills). One of the therapeutic agents may be given in multiple doses, or both may be given as multiple doses. In addition, the combination methods, compositions and formulations are not to be limited to the use of only two agents.

VII. Kits for Treatment or Prevention of CIPN

[0066] The following discussion is presented by way of example only, and is not intended to be limiting. [0067] In some embodiments, the present disclosure provides for a kit useful for treating or preventing CIPN comprising a composition comprising a bacterial strain selected from the group consisting of ART 12 (NCIMB Accession No. 43087) and ART24 (NCIMB Accession No. 43088), and a package insert with instructions for treating or preventing CIPN in a subject in need thereof. In some embodiments, the composition comprises ART12, ART24, or both ART12 and ART24. In some embodiments, the bacterial strain(s) is concentrated at about 1x10⁸ colony forming units (CFU) per ml to about 1x10¹² CFU per ml of ART24 and/or ART12 bacterium (i.e., vegetative cell ) or bacterial spore. In some embodiments, the package insert instructs that about 1x10⁹ to about 1x10¹² viable bacteria or spores be administered per day. In some embodiments, the compositions of the kit are delivered as lyophilized material or powder to be re-suspended for oral delivery or packaged into capsules. The lyophilized material and capsules may be coated for improved enteric stability. In some embodiments, the bacterial strain(s) is lyophilized. The compositions of the kit may be formulated in effective amounts (i.e., amounts that have desired therapeutic effect). The dose and dosage regimen will depend upon the degree of CIPN progression and symptom severity in the subject, the characteristics of the ART 12 and/or ART24 strain used, e.g., its therapeutic index, the subject, and the subject’s history. The effective amount may be determined during pre-clinical trials and clinical trials by methods familiar to physicians and clinicians, and instructions regarding the effective amount can be included on the package insert.

[0068] In some embodiments, the kit further comprises a co-therapeutic. In some embodiments, the co-therapeutic comprises one or more disease modifying agents, such as, but not limited to, those described herein. In some embodiments, the package insert instructs that the co-therapeutic be provided simultaneously, sequentially, or separately to the bacterial strain(s). In some embodiments, the kit further comprises other materials useful for treating CIPN, such as containers (e.g., bottles, vials, syringes, etc}, additional buffers or solutions (e.g., saline, PBS, or cleaning solutions), additional instructions regarding combination therapies, and memory aids (e.g., a calendar insert to remind a subject to take a dose as instructed). [0069] In some embodiments, the package insert instructs that the bacterial strain(s) be administered daily, twice daily, or three times daily. In some embodiments, the package insert instructs that the bacterial strain(s) be administered orally, buccaly, rectally, or sublingually. In some embodiments, the package insert instructs that the bacterial strain(s) be added to a food or beverage. In some embodiments, the package insert i nstructs that the CIPN symptoms of a subject receiving the bacterial strain(s) be tracked during the period of treatment.

EXAMPLES

[0070] The following examples are provided by way of illustration only and not by way of limitation. Those of skill in the art will readily recognize a variety of non-critical parameters that could be changed or modified to yield essentially the same or similar results. The examples should in no way be construed as limiting the scope of the present technology, as defined by the appended claims.

Example 1: ART12 and ART24 in Paclitaxel-Induced Peripheral Neuropathy.

Animal Model

[0071] This example demonstrates that ART12 and ART24 are effective in methods for treating or preventing CIPN in an in vivo mouse model.

Introduction

[0072] Paclitaxel (Taxol®) is classified as a plant alkaloid, a taxane, and an antimicrotubule agent. Paclitaxel is widely used in the treatment of cancers such as breast, ovarian, lung, bladder, prostate, melanoma, esophageal, as well as other types of solid tumor cancers. It has also been used in Kaposi’s sarcoma. Although highly effective in blocking tumor progression, paclitaxel is also associated with the development of peripheral neuropathy as a side effect in approximately 60-70% of chemotherapy patients. Summary and Procedures

[0073] The aim of this study was to evaluate the efficacy of prophylactic treatment with ART 12 or ART24 in the Taxol® induced neuropathic pain model in mice, using von Frey test for mechanical allodynia. Taxol® was administered intraperitoneally (IP), at a dose level of 6 mg/kg, for 8 consecutive days (Days 0-7), to animals allocated into groups 2-6. Animals in group 1 were not treated with Taxol. Animals in groups 1 and 2 received treatment with Placebo, animals in group 3 received treatment with ART24 (Lot# MP-FD- 230920; Lot#15), at a dose level of ~5.00E+08/dose, and animals in group 4 received treatment with Cryo. Treatment was twice daily, starting from study day -3 until day 14. Animals in group 5 were treated with ART 12 (Lot# MP-FD-301120), at a dose level of ~5.00E+08/dose, twice daily from study day -3 until day -1, while on study days 0 until 14, these animals were treated with ART12 (Lot# MP-FD-030221), at a dose level of ~3.70E+08/dose. Gabapentin, which was used as the positive control, was administered to animals in group 6, on testing days, 8 and 14, 2 hours prior to von Frey test. Von Frey was performed at baseline (study day -4), and then on study days 8 and 14. A schematic depiction of the paclitaxel (Taxol® ) induction and treatment procedure is shown in Figure 1A.

[0074] Experimental items. Details of materials are provided in Table 1.

Table 1. Experimental items.

[0075] Constitution of test groups and dose levels. The six experimental groups in this study are described in Table 2. Table 2. Experimental gronps.

[0076] Von Frey Measurements. Von Frey measurements were conducted on study days - 4 (baseline) and then on study days 8 and 14.

[0077] Statistics/ 'Data evaluation. All data are presented as Mean±SEM. Each treatment group was compared to the Placebo group (Group 2) using Student’s T-test, or one-way ANOVA, followed by Dunnett’s multiple comparisons test. A p value < 0.05 is considered to represent a significant difference.

[0078] Animal care and use statement. This study was performed following approval of an application form submitted to the Committee for Ethical Conduct in the Care and Use of Laboratory Animals that stated that the study complied with the rules and regulations set forth.

Results

Von Frey Measurements

[0079] As shown in Figure 1B and Table 3, animals treated with the placebo (Group 2) showed a statistically significant reduction in the mean withdrawal force threshold at all testing days (days 8 and 14), compared to the placebo/without Taxol® group (group 1): 0.20 ±0.03g vs. 1.73±0.09g for the placebo/without Taxol® group, on study day 14; p<0.0001, using one-way ANOVA, followed by Dunnett’s post-hoc test. Animals treated with Gabapentin, at a dose level of 100 mg/kg PO (group 6), the positive control, showed a statistically significantly higher mean withdrawal force threshold on all testing days (days 8 and 14), 2 hours post administration, compared to the placebo group: 1.61±0.16g vs.

0.20±0.03g for the placebo group, on study day 14, 2 hours post administration, p<0.0001, using one-way ANOVA, followed by Dunnett’s post-hoc test (Figure IB; Table 3). Also as shown in Figure 1B and Table 3, treatment with ART24 (Group 3) or ART12 (Group 5), resulted in a statistically significant higher mean withdrawal force threshold, on study day 14, compared to the placebo group: 0.33±0.05g for ART24 or 0.46±0.08g for ART12 vs. 0.20±0.03g for the placebo group, on study day 14; p<0.05 using T-test. Table 3. Mean group response to von Frey (g).

#p<0.05 vs. Placebo (Group 2) using Student’s T-test

##p<0.01 vs. Placebo (Group 2) using Student’s T-test

****p<() .0001 vs. Placebo (Group 2) using one-way ANOVA, followed by Dunnett’s post- hoc test.

Conclusion

[0080] Under the conditions of this study and confined to the in-life data, treatment with ART24 (Group 3) or ART 12 (Group 5), resulted in a statistically significant slight reduction in mechanical allodynia, induced by administration of Taxol®, when administered twice daily from study day -3-14. Accordingly, these results demonstrate that the ART12 and ART24 strains of the present technology are effective in methods for treating or preventing CIPN in a subject in need thereof. Human Subjects

[0081] Human subjects diagnosed as having or suspected to have CIPN and presently displaying one or more symptoms and/or pathologies of CIPN, are recruited using selection criteria known and accepted in the art.

Methods of Prevention and Treatment

[0082] Subjects are administered ART12 and/or ART24 at a dosage and frequency commensurate with the stage and severity of disease. In some embodiments ART 12 and/or ART24 is administered once daily, once weekly, or once monthly. In some embodiments, ART 12 and/or ART24 is administered multiple times daily, weekly, or monthly.

[0083] To demonstrate methods of prevention and treatment in humans, subjects are administered ART12 and/or ART24 prior to or subsequent to the development of symptoms and/or pathologies of CIPN and assessed for reversal of symptoms/pathologies or attenuation of expected symptoms/pathologies using methods known in the art.

Results

[0084] It is expected that ART 12 and/or ART24 will inhibit the worsening of symptoms and/or pathologies of CIPN in human subjects. These results will show that ART 12 and/or ART24 is useful and effective for the prevention and treatment of such CIPN.

Example 2: ART12 and ART24 in Cisplatin-Induced Peripheral Neuropathy

Animal Model

[0085] This example demonstrates that ART12 and ART24 are effective in methods for treating or preventing CIPN in an in vivo mouse model.

Introduction

[0086] Cisplatin is a platinum-based chemotherapeutic that is commonly used for the treatment of solid tumors such as lung, ovarian, testis, bladder, and head and neck cancer. Cisplatin treatment is associated with a high incidence of chemotherapy-induced peripheral neuropathy (CIPN), and in rodents it induces mechanical allodynia, spontaneous pain, and numbness. Mechanisms that underlie the development and maintenance of cisplatin- induced peripheral neuropathy are under active investigation.

Summary and Procedures

[0087] The aim of this study is to evaluate the efficacy of ART12 and ART24 in the cisplatin-induced neuropathic pain model in mice. Neuropathic pain was induced in mice using cisplatin (2.3 mg/kg), administered intraperitoneally (IP), for 5 consecutive days (0- 4), followed by 5 days off (5-9), and then administered for an additional 5 consecutive days (10-14). Animals in groups 2-5 were injected with cisplatin, while animals in Group 1 were injected with saline.

[0088] The development of neuropathic pain was confirmed by measuring mechanical allodynia (von Frey test). Animals were treated PO with ART12 (Group 3) or ART24 (Group 4), at the dose levels of IxlO⁹ CFU, or with the vehicle (PBS, Groups 1 and 2), 150 mg/kg gabapentin (IP, Group 5) served as a positive control. A schematic depiction of the cisplatin induction and treatment procedure is shown in Figure 2A.

[0089] Experimental items. Details of materials are provided in Table 4.

Table 4. Experimental items.

RT = room temperature; N/A not applicable

[0090] Constitution of test groups and dose levels. The five experimental groups in this study are described in Table 5.

Table 5. Experimental groups.

[0091] Von Frey Measurements. Mechanical pain sensitivity was tested using the von Frey test, which measured the withdrawal force threshold of the animals. Von Frey test was performed on day -1 (baseline) and on days 15 and 18 at 2 hours post dosing.

[0092] Statistics/Data evaluation. All data are presented as Mean±SEM. Each treatment group was compared to the cisplatin + PBS group (Group 2) using one-way ANOVA followed by Dunnett’s post-test (GraphPad). A p-value < 0.05 is considered to represent a significant difference.

[0093] Animal care and use statement. This study was performed following approval of an application form submitted to the Committee for Ethical Conduct in the Care and use of Laboratory Animals that stated that the study complied with the rules and regulations set forth.

Results

Von Frey Measurements

[0094] As shown in Figure 2B and Table 6, treatment with ART12 (Group 3) or ART24 (Group 4) resulted in a statistically significantly higher withdrawal force response on testing days 15 and 18. Data for the individual animals is provided in Table 7. Thus, on study day 15, the withdrawal force was 0.78±0.09g (Group 3), 0.83±0. 11g vs. 0.18±0.01g for cisplatin + PBS treated animals (Group 2); p<0.0001 using one way ANOVA test followed by Dunnett’s test (day 15). On study day 18 the withdrawal force was 0.80±0.09g (Group 3), 0.75±0.12g (Group 4) vs. 0.18±0.01g of cisplatin + PBS treated animals (Gorup 2); p<0.0001 using one way ANOVA test followed by Dunnett’s test.

Table 6. Mean group response to von Frey (g).

p<0.0001 vs. Cisplatin+PBS (Group 2) using a one-way ANOVA test followed by Dunnett’s test. p<0.0001 vs. Saline • PBS (Group 1) using student’s T-test.

Table 7. Individual animal data in response to von Frey (g).

R = Right leg; L = Left leg; FDC = found dead in cage.

Conclusion

[0095] Treatment of subjects with ART12 (Group 3) or ART24 (Group 4) resulted in a statistically significant reduction of neuropathic pain caused by cisplatin administration as measured by the von Frey test. Accordingly, these results demonstrate that the ART 12 and ART24 strains of the present technology are effective in methods for treating or preventing CIPN in a subject in need thereof.

Human Subjects

[0096] Human subjects diagnosed as having or suspected to have CIPN and presently displaying one or more symptoms and/or pathologies of CIPN, are recruited using selection criteria known and accepted in the art.

Methods of Prevention and Treatment

[0097] Subjects are administered ART12 and/or ART24 at a dosage and frequency commensurate with the stage and severity of disease. In some embodiments ART 12 and/or ART24 is administered once daily, once weekly, or once monthly. In some embodiments, ART12 and/or ART24 is administered multiple times daily, weekly, or monthly.

[0098] To demonstrate methods of prevention and treatment in humans, subjects are administered ART12 and/or ART24 prior to or subsequent to the development of symptoms and/or pathologies of CIPN and assessed for reversal of symptoms/pathol ogies or attenuation of expected symptoms/pathologies using methods known in the art.

Results

[0099] It is expected that ART 12 and/or ART24 will inhibit the worsening of symptoms and/or pathologies of CIPN in human subjects. These results will show that ART12 and/or ART24 is useful and effective for the prevention and treatment of such CIPN.

BIOLOGICAL DEPOSITS

[0100] The Applicant requests that a sample of the deposited microorganisms should be made available only to an expert approved by the Applicant.

[0101] Bacillus amyloliquefaciens strain ART12 deposited with the National Collection of Industrial Food and Marine Bacteria (NCIMB Ltd.) (International Depositary Authority), Ferguson Building, Craibstone Estate, Bucksbum, Aberdeen, AB21 9YA, Scotland, on 21 June 2018, under NCIMB Accession Number 43087.

[0102] Bacillus amyloliquefaciens strain ART24 deposited with the National Collection of Industrial Food and Marine Bacteria (NCIMB Ltd.) (International Depositary' Authority), Ferguson Building, Craibstone Estate, Bucksbum, Aberdeen, AB21 9YA, Scotland, on 21 June 2018, under NCIMB Accession Number 43088.

[0103] The deposits were made according to the Budapest treaty on the international recognition of the deposit of microorganisms for the purposes of patent procedure.

EQUIVALENTS

[0104] The present technology is not to be limited in terms of the particular embodiments described in this application, which are intended as single illustrations of individual aspects of the present technology. Many modifications and variations of this present technology can be made without departing from its spirit and scope, as will be apparent to those skilled in the art. Functionally equivalent methods and apparatuses within the scope of the present technology, in addition to those enumerated herein, will be apparent to those skilled in the art from the foregoing descriptions. Such modifications and variations are intended to fall within the scope of the appended claims. The present technology is to be limited only by the terms of the appended claims, along with the full scope of equivalents to which such claims are entitled. It is to be understood that this present technology is not limited to particular methods, reagents, compounds compositions or biological systems, which can, of course, vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting.

[0105] Each and every publication and patent mentioned in the above specification is herein incorporated by reference in its entirety for all purposes. Various modifications and variations of the described methods and system of the present technology will be apparent to those skilled in the art without departing from the scope and spirit of the present technology. Although the present technology has been described in connection with specific embodiments, the present technology as claimed should not be unduly limited to such specific embodiments. Indeed, various modifications of the described modes for carrying out the present technology which are obvious to those skilled in the art and in fields related thereto are intended to be within the scope of the following claims.

Claims

What is claimed is:

1. A method for treating or preventing chemotherapy -induced peripheral neuropathy (CIPN) in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising bacterial strain ART12 (NCIMB Accession No. 43087), ART24 (NCIMB Accession No. 43088), or a combination thereof.

2. The method of claim 1, wherein the bacterial strain is lyophili zed.

3. The method of claim 1 or claim 2, wherein the bacterial strain is in the form of a spore.

4. The method of claim 1 or claim 2, wherein the bacterial strain is in vegetative form.

5. The method of claim 1 or claim 2, wherein the bacterial strain is a mixture of the strain in the form of a spore and in the vegetative form.

6. The method of any one of claims 1-5, wherein the bacterial strain is formulated as a dietary supplement.

7. The method of any of claims 1-5, wherein the bacterial strain is formulated as an edible product.

8. The method of claim 7, wherein the edible product further comprises a carrier, vehicle, or excipient.

9. The method of claim 7, wherein the edible product further compri ses a fermented food product, soybean, mushroom, mung bean, locus bean, rice, or extracts thereof.

10. The method of claim 9, wherein the soybean is fermented soybean or fermented soybean paste. The method of claim 10, wherein the fermented soybean or fermented soybean paste is Cheonggukjang, Douchi, Hawaijar, Bekang, Peruyaan, Tungrymbai, Eoyukjang, Kinema, Aakhone, Miso, Natto, or Thua-nao. The method of any one of claims 1-11, wherein the CIPN comprises one or more of myalgias; paresthesia; glove-and- stocking sensory neuropathy; hyperalgia; allodynia; sharp, stabbing pain, burning or shock-like sensations; loss of sensation or complete numbness; trouble with small motor skills; gripping problems; clumsiness; weakness; and balance problems. The method of any one of claims 1-12, wherein treatment or prevention comprises reducing allodynia. The method of any one of claims 1 -13, wherein the pharmaceutical composition is administered orally. The method of any one of claims 1-14, further comprising separately, sequentially, or simultaneously administering one or more agents selected from the group consisting of a steroid, lidocaine, capsaicin, an anti-seizure medication, an antidepressants, an analgesic, and any combination thereof. The method of any one of claims 1-15, wherein the CIPN is associated with paclitaxel. The method of any one of claims 1-15, wherein the CIPN is associated with cisplatin. Use of a composition in the preparation of a medicament for treating or preventing chemotherapy-induced peripheral neuropathy (CIPN) in a subject in need thereof, wherein the composition comprises bacterial strain ART12 (NCIMB Accession No. 43087), ART24 (NCIMB Accession No. 43088), or a combination thereof The use of claim 18, wherein the bacterial strain is lyophilized. The use of claim 18 or claim 19, wherein the bacterial strain is in the form of a spore. The use of claim 18 or claim 19, wherein the bacterial strain is in vegetative form. The use of claim 18 or claim 19, wherein the bacterial strain is a mixture of the strain in the form of a spore and in the vegetative form. The use of any one of claims 19-22, wherein the bacterial strain is formulated as a dietary supplement. The use of any of claims 19-22, wherein the bacterial strain is formulated as an edible product. The use of claim 24, wherein the edible product further comprises a carrier, vehicle, or excipient. The use of claim 24, wherein the edible product further comprises a fermented food product, soybean, mushroom, mung bean, locus bean, rice, or extracts thereof. The use of claim 26, wherein the soybean is fermented soybean or fermented soybean paste. The use of claim 27, wherein the fermented soybean or fermented soybean paste is Cheonggukjang, Douchi, Hawaijar, Bekang, Peruyaan, Tungrymbai, Eoyukjang, Kinema, Aakhone, Miso, Natto, or Thua-nao. The use of any one of claims 18-28, wherein the CIPN comprises one or more of myalgias; paresthesia; glove-and-stocking sensory neuropathy; hyperalgia; allodynia; sharp, stabbing pain; burning or shock-like sensations; loss of sensation or complete numbness; trouble with small motor skills; gripping problems; clumsiness; weakness; and balance problems. The use of any one of claims 18-29, wherein treatment or prevention comprises reducing allodynia. The use of any one of claims 18-30, wherein the pharmaceutical composition is formulated for oral administration. The use of any one of claims 18-31, further comprising separately, sequentially, or simultaneously administering one or more agents selected from the group consisting of a steroid, lidocaine, capsaicin, an anti-seizure medication, an antidepressants, an analgesic, and any combination thereof. The use of any one of claims 18-32, wherein the CIPN is associated with paclitaxel. The use of any one of claims 18-32, wherein the CIPN is associated with cisplatin. A composition used for treating or preventing chemotherapy-induced peripheral neuropathy (CIPN) in a subject in need thereof, wherein the composition comprises bacterial strain ART12 (NCIMB Accession No. 43087), ART24 (NCIMB Accession No. 43088), or a combination thereof. The composition for use of claim 35, wherein the bacterial strain is lyophilized. The composition for use of claim 35 or claim 36, wherein the bacterial strain is in the form of a spore. The composition for use of claim 35 or claim 36, wherein the bacterial strain is in vegetative form. The composition for use of claim 35 or claim 36, wherein the bacterial strain is a mixture of the strain in the form of a spore and in the vegetative form. The composition for use of any one of claims 36-39, wherein the bacterial strain is formulated as a dietary supplement. The composition for use of any of claims 36-39, wherein the bacterial strain is formulated as an edible product. The composition for use of claim 41 , wherein the edible product further comprises a earner, vehicle, or excipient. The composition for use of claim 41, wherein the edible product further comprises a fermented food product, soybean, mushroom, mung bean, locus bean, rice, or extracts thereof. The composition for use of claim 43, wherein the soybean is fermented soybean or fermented soybean paste. The composition for use of claim 44, wherein the fermented soybean or fermented soybean paste is Cheonggukjang, Douchi, Hawaijar, Bekang, Peruyaan, Tungrymbai, Eoyukjang, Kineraa, Aakhone, Miso, Natto, or Thua-nao. The composition for use of any one of claims 35-45, wherein the CIPN comprises one or more of myalgias; paresthesia; glove-and-stocking sensory' neuropathy; hyperalgia; allodynia; sharp, stabbing pain; burning or shock-like sensations, loss of sensation or complete numbness; trouble with small motor skills; gripping problems; clumsiness; weakness; and balance problems. The composition for use of any one of claims 35-46, wherein treatment or prevention comprises reducing allodynia. The composition for use of any one of claims 35-47, wherein the pharmaceutical composition is formulated for oral administration. The composition for use of any one of claims 35-48, further comprising separately, sequentially, or simultaneously administering one or more agents selected from the group consisting of a steroid, lidocaine, capsaicin, an anti-seizure medication, an antidepressants, an analgesic, and any combination thereof. The composition for use of any one of claims 35-49, wherein the CIPN is associated with paclitaxel. The composition for use of any one of claims 35-49, wherein the CIPN is associated with cisplatin. A kit for use in treating or preventing CIPN comprising:

(a) a composition comprising a bacterial strain selected from the group consisting of: ART12 (NCIMB Accession No. 43087) and ART24 (NCIMB Accession No. 43088); and

(b) a package insert with instructions for treating or preventing CIPN in a subject in need thereof. The kit of claim 52, further comprising a co-therapeutic selected from the group consisting of a steroid, lidocaine, capsaicin, an anti-seizure medication, an antidepressants, an analgesic, and any combination thereof.