Oral gemfibrozil for the treatment of cln3-related juvenile neuronal ceroid lipofuscinosis

Abstract

The present disclosure generally relates to improved pharmaceutical compositions useful for the treatment of juvenile neuronal ceroid lipofuscinosis (JNCL) or Juvenile Batten Disease which is a fatal inherited neurodegenerative disease of children caused by impairments in Cln3 gene function. More particularly, the disclosure relates to pharmaceutical compositions comprising oral gemfibrozil for the treatment of juvenile neuronal ceroid lipofuscinosis (JNCL) or Juvenile Batten Disease.

Description

ORAL GEMFIBROZIL FOR THE TREATMENT OF CLN3-RELATED JUVENILE NEURONAL CEROID LIPOFUSCINOSIS

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This applications claims priority to U.S. Provisional Application Serial No. 63 / 439,723 filed January 18, 2023. The entire contents are incorporated herein by reference.

[0003] GOVERNMENT LICENSE RIGHTS CLAUSE

[0004] This invention was made with government support under AG050431 and AT010980 awarded by the National Institutes of Health and under 1K6BX004982 awarded by the United States Department of Veterans Affairs. The government has certain rights in the invention

[0005] SEQUENCE LISTING

[0006] A sequence listing prepared under the WIPO Standard ST.26 was prepared on January' 17, 2024, and having a file size of -7,000 bytes, is a part of and submitted herewith this disclosure.

[0007] FIELD OF THE INVENTION

[0008] The present disclosure generally relates to methods and pharmaceutical compositions useful for the treatment of juvenile neuronal ceroid lipofuscinosis (JNCL) also referred to as Batten Disease, which is a fatal inherited neurodegenerative disease of children caused by mutations in the molecular pathway or directly in the Cln3 gene. More particularly, the disclosure relates to methods of treatments and pharmaceutical compositions comprising oral gemfibrozil for the treatment of JNCL.

[0009] BACKGROUND OF THE INVENTION

[0010] Juvenile neuronal ceroid lipofuscinosis (JNCL), or Batten disease, is a fatal autosomal recessive inherited childhood-onset neurodegenerative disorder caused by mutations in the Cln3 gene. With an estimated occurrence of 1 in 12,500 live births, JNCL is considered a rare neurological disease that usually begins between 4 and 10 years of age, with progressive visual deterioration, seizures, blindness, motor and cognitive decline, mental and intellectual deterioration, epilepsy, and ultimately ending with premature death during the second and third decade of life. Despite intense investigations, no effective therapy is available for JNCL.

[0011] Gemfibrozil is an FDA-approved lipid-lowering drug prescribed for its ability to reduce the level of plasma triglycerides and decrease the risk of hyperlipidemia in patients. Gemfibrozil, commonly known as “Lopid,” was successfully introduced on the market in 1976.

[0012] With this background in mind, the disclosure below describes a possible therapeutic use of gemfibrozil in JNCL patients.

[0013] SUMMARY OF THE INVENTION

[0014] Baten Disease appears to be caused, at least in part, by two mutated copies of the Cln3 gene. The disclosure herein utilizes the discovery that gemfibrozil treatment reduced microglial and astroglial activation, atenuated neuroinflammation, restored the level of TFEB (the master regulator of lysosomal biogenesis), and decreased the accumulation of storage material containing subunit c of mitochondrial ATP synthase (SCMAS) in somatosensory barrel field (SBF) cortex of a JNCL mouse model. Without being limited by theory, it is believed that molecular dysfunction caused by at least one mutation in the both copies of the Cln3 gene can be rescued or alleviated through the introduction of gemfibrozil, which stimulates recruitment of PPARa to the Tfeb gene promoter in vivo in the SBF cortex. Additionally, the introduction of gemfibrozil reduced microglial and astroglial activation, which reduced neuroinflammation; increased the level of TFEB, which is lower in JNCL patients; and decreased the accumulation of SCMAS in the SBF. Further, oral treatment in the JNCL mouse models showed an improvement in locomotor function. Taken together, this data suggests that gemfibrozil can be used as a continuous treatment for patients diagnosed or suspected of having JNCL.

[0015] JNCL features glial activation and accumulation of autofluorescent storage material containing subunit c of mitochondrial ATP synthase (SCMAS), ultimately resulting into neuronal loss. Until now, no effective therapy is available for JNCL. This study underlines the possible therapeutic importance of gemfibrozil, a prototype activator of peroxisome proliferator-activated receptor a (PPARa) and food and drug administration (FDA)-approved lipid-lowering drug, in an animal model of JNCL. Oral gemfibrozil treatment reduced microglial and astroglial activation, attenuated neuroinflammation, restored the level of TFEB (the master regulator of lysosomal biogenesis), and decreased the accumulation of storage material SCMAS in somatosensory barrel field (SBF) cortex of Cln3Aex7 / 8(Cln3AJNCL) mice. Accordingly, gemfibrozil treatment also improved locomotor activities of Cln3AJNCL mice. While investigating the mechanism, we found marked loss of PPARa in the SBF cortex of Cln3AJNCL mice, which increased after gemfibrozil treatment. Oral gemfibrozil also stimulated the recruitment of PPARa to the Tfeb gene promoter in vivo in the SBF cortex of Cln3AJNCL mice, indicating increased transcription of Tfeb in the CNS by gemfibrozil treatment via PPARa. Moreover, disease pathologies aggravated in Cln3AJNCL mice lacking PPARa (Cln3AJNCLAPPARra) and gemfibrozil remained unable to decrease SCMAS accumulation, reduce glial activation and improve locomotor performance ofCln3AJNCLP,'AR" mice. These results suggest that activation of PPARa may be beneficial for JNCL and that gemfibrozil may be repurposed for the treatment of this incurable disease.

[0016] The present investigation was carried out to examine whether gemfibrozil could halt and / or slow down the disease process of JNCL in Cln3Aex7 / 8(Cln3AJNCL) mice. Here, the inventors provide evidence that orally administered gemfibrozil markedly attenuated the glial activation, increased the level of TFEB to reduce the accumulation of SCMAS, and improved locomotor activities in Cln3AJNCLAPPARamice via PPARa. Here, we delineate that oral administration of gemfibrozil, a lipid-lowering drug, decreases glial inflammation, normalizes and / or upregulates TFEB and reduces accumulation of autofluorescent storage material in SBF cortex to improve locomotor activities in Cln3Aex7 / 8(Cln3AJNCL) mice. Aggravation of disease pathology in Cln3AJNCL mice lacking PPARa (Cln3AJNCLAPPARa) and inability of gemfibrozil to decrease SCMAS accumulation, reduce glial activation and improve locomotor performance of Cln3AJNCLAPPAR“ mice delineates an important role of PPARa in this process.

[0017] In various embodiments, the present disclosure provides methods for treating Juvenile neuronal ceroid lipofuscinosis (JNCL) in a patient in need thereof.

[0018] In other embodiments, are provided methods for treating or correcting a defective function of the lysosomal membrane glycoprotein Cln3.

[0019] In any embodiment the methods comprise administering to a patient in need thereof an effective amount of an oral pharmaceutical composition comprising gemfibrozil. Gemfibrozil may be identified by its CAS identifier 25812-30-0.

[0020] In any of the disclosed embodiments, the pharmaceutical composition may be administered to the patient in any including one time per day, two times per day, and three times per day.

[0021] The foregoing has outlined rather broadly the features and technical advantages of the present disclosure in order that the detailed description that follows may be better understood. Additional features and advantages of the disclosure will be described hereinafter that form the subject of the claims of this application. It should be appreciated by those skilled in the art that the conception and the specific embodiments disclosed may be readily utilized as a basis for modifying or designing other embodiments for carrying out the same purposes of the present disclosure. It should also be realized by those skilled in the art that such equivalent embodiments do not depart from the spirit and scope of the disclosure as set forth in the appended claims.

[0022] BRIEF DESCRIPTION OF THE DRAWINGS

[0023] FIG. 1A shows a graph depicting the amount of GFAP relative to WT-control with the first bar closest to the y-axis representing WT, the second bar representing the JNCL mouse model, the third bar representing the mouse model dosed with 4mg / kg / d, and the fourth bar the mouse model dosed with 8mg / kg / d.

[0024] FIG. IB shows a graph depicting the amount of Ibal relative to WT-control with the first bar closest to the y-axis representing WT, the second bar representing the JNCL mouse model, the third bar representing the mouse model dosed with 4mg / kg / d, and the fourth bar the mouse model dosed with 8mg / kg / d.

[0025] FIG. 1C shows a graph depicting the number of GFAP -positive cells with the first bar closest to the y-axis representing WT, the second bar representing the JNCL mouse model, the third bar representing the mouse model dosed with 4mg / kg / d, and the fourth bar the mouse model dosed with 8mg / kg / d.

[0026] FIG. ID shows a graph depicting the number of Iba-1 -positive cells with the first bar closest to the y-axis representing WT, the second bar representing the JNCL mouse model, the third bar representing the mouse model dosed with 4mg / kg / d, and the fourth bar the mouse model dosed with 8mg / kg / d.

[0027] FIG. 2A shows a graph depicting the amount of iNOS relative to WT-control with the first bar closest to the y-axis representing WT, the second bar representing the JNCL mouse model, the third bar representing the mouse model dosed with 4mg / kg / d, and the fourth bar the mouse model dosed with 8mg / kg / d.

[0028] FIG. 2B shows a graph depicting the amount of Pro IL- 1 (3 relative to WT- control with the first bar closest to the y-axis representing WT, the second bar representing the JNCL mouse model, the third bar representing the mouse model dosed with 4mg / kg / d, and the fourth bar the mouse model dosed with 8mg / kg / d.

[0029] FIG. 2C shows a graph depicting the amount of mature IL- 1(3 relative to WT- control with the first bar closest to the y-axis representing WT, the second bar representing the JNCL mouse model, the third bar representing the mouse model dosed with 4mg / kg / d, and the fourth bar the mouse model dosed with 8mg / kg / d.

[0030] FIG. 2D shows a graph depicting the amount of TNF-a relative to WT-control with the first bar closest to the y-axis representing WT, the second bar representing the JNCL mouse model, the third bar representing the mouse model dosed with 4mg / kg / d, and the fourth bar the mouse model dosed with 8mg / kg / d.

[0031] FIG. 3A shows a graph depicting the amount of MFI of neuronal TFEB with the first bar closest to the y-axis representing WT, the second bar representing the JNCL mousemodel, the third bar representing the mouse model dosed with 4mg / kg / d, and the fourth bar the mouse model dosed with 8mg / kg / d.

[0032] FIG. 3B shows a graph depicting the amount of MFI of NeuN with the first bar closest to the y-axis representing WT, the second bar representing the JNCL mouse model, the third bar representing the mouse model dosed with 4mg / kg / d, and the fourth bar the mouse model dosed with 8mg / kg / d.

[0033] FIG. 3C shows a graph depicting the amount of TFEB relative to WT-control with the first bar closest to the y-axis representing WT, the second bar representing the JNCL mouse model, the third bar representing the mouse model dosed with 4mg / kg / d, and the fourth bar the mouse model dosed with 8mg / kg / d.

[0034] FIG. 3D shows a graph depicting the amount of MFI of SCMAS with the first bar closest to the y-axis representing WT, the second bar representing the JNCL mouse model, the third bar representing the mouse model dosed with 4mg / kg / d, and the fourth bar the mouse model dosed with 8mg / kg / d.

[0035] FIG. 4A shows a graph depicting the amount of TPP1 relative to WT-control with the first bar closest to the y-axis representing WT. the second bar representing the JNCL mouse model, the third bar representing the mouse model dosed with 8mg / kg / d.

[0036] FIG. 4B shows a graph depicting the amount of p62 relative to WT-control with the first bar closest to the y-axis representing WT, the second bar representing the JNCL mouse model, the third bar representing the mouse model dosed with 8mg / kg / d.

[0037] FIG. 4C shows a graph depicting the amount of MFI of neuronal TPP1 with the first bar closest to the y-axis representing WT, the second bar representing the JNCL mouse model, the third bar representing the mouse model dosed with 8mg / kg / d.

[0038] FIG. 4D shows a graph depicting the amount of MFI of neuronal p62 with the first bar closest to the y-axis representing WT, the second bar representing the JNCL mouse model, the third bar representing the mouse model dosed with 8mg / kg / d.

[0039] FIG. 5A shows a graph depicting the velocity (cm / sec) with the first bar closest to the y-axis representing WT, the second bar representing the JNCL mouse model, the third bar representing the mouse model dosed with 4mg / kg / d, and the fourth bar the mouse model dosed with 8mg / kg / d.

[0040] FIG. 5B shows a graph depicting the cumulative duration in seconds with the first bar closest to the y-axis representing WT, the second bar representing the JNCL mouse model, the third bar representing the mouse model dosed with 4mg / kg / d, and the fourth bar the mouse model dosed with 8mg / kg / d.

[0041] FIG. 5C shows a graph depicting the distance traveled in centimeters with the first bar closest to the y-axis representing WT, the second bar representing the JNCL mouse model, the third bar representing the mouse model dosed with 4mg / kg / d, and the fourth bar the mouse model dosed with 8mg / kg / d.

[0042] FIG. 5D shows a graph depicting the center point moving in seconds with the first bar closest to the y-axis representing WT, the second bar representing the JNCL mouse model, the third bar representing the mouse model dosed with 4mg / kg / d, and the fourth bar the mouse model dosed with 8mg / kg / d.

[0043] FIG. 5E shows a graph depicting the pole latency in seconds with the first bar closest to the y-axis representing WT, the second bar representing the JNCL mouse model, the third bar representing the mouse model dosed with 4mg / kg / d. and the fourth bar the mouse model dosed with 8mg / kg / d.

[0044] FIG. 5F shows a graph depicting the rotorod latency in seconds with the first bar closest to the y-axis representing WT, the second bar representing the JNCL mouse model, the third bar representing the mouse model dosed with 4mg / kg / d, and the fourth bar the mouse model dosed with 8mg / kg / d.

[0045] FIG. 6A shows a graph depicting the amount of PPARa relative to WT-control with the first bar closest to the y-axis representing WT, the second bar representing the JNCL mouse model, the third bar representing the mouse model dosed with 8mg / kg / d.

[0046] FIG. 6B shows a graph depicting the amount of MFI of PPARa with the first bar closest to the y-axis representing WT, the second bar representing the JNCL mouse model, the third bar representing the mouse model dosed with 8mg / kg / d.

[0047] FIG. 6C shows a schematic representation of the mouse Tfeb gene promoter with PPRE.

[0048] FIG. 6D shows a graph depicting the quantitative PCR of PPARa, CBP, RNA Pol, IgG, and Input (control) with the first bar closest to the y-axis representing WT, the second bar representing the JNCL mouse model, the third bar representing the mouse model dosed with 8mg / kg / d.

[0049] FIG. 7 A shows a graph depicting the amount of Ibal relative to WT-control with the first bar closest to the y-axis representing WT, the second bar representing the JNCL mouse model, the third bar representing JNCL and PPARa null model.

[0050] FIG. 7B show s a graph depicting the amount of GFAP relative to WT-control with the first bar closest to the y-axis representing WT, the second bar representing the JNCL mouse model, the third bar representing JNCL and PPARa null model.

[0051] FIG. 7C shows a graph depicting the amount of MFI of SCMAS with the first bar closest to the y-axis representing WT, the second bar representing the JNCL mouse model, the third bar representing JNCL and PPARa null model.

[0052] FIG. 7D shows a graph depicting the pole T-tum in seconds with the first bar closest to the y-axis representing WT, the second bar representing the JNCL mouse model, the third bar representing JNCL and PPARa null model.

[0053] FIG. 7E shows a graph depicting the pole latency in seconds with the first bar closest to the y-axis representing WT, the second bar representing the JNCL mouse model, the third bar representing JNCL and PPARa null model.

[0054] FIG. 7F shows a graph depicting the rotorod test in seconds with the first bar closest to the y-axis representing WT, the second bar representing the JNCL mouse model, the third bar representing JNCL and PPARa null model.

[0055] FIG. 7G shows a graph depicting the velocity in centimeters per seconds with the first bar closest to the y-axis representing WT, the second bar representing the JNCL mouse model, the third bar representing JNCL and PPARa null model.

[0056] FIG. 7H shows a graph depicting the cumulative duration in seconds with the first bar closest to the y-axis representing WT, the second bar representing the JNCL mouse model, the third bar representing JNCL and PPARa null model.

[0057] FIG. 71 shows a graph depicting the distance traveled in centimeters with the first bar closest to the y-axis representing WT, the second bar representing the JNCL mouse model, the third bar representing JNCL and PPARa null model.

[0058] FIG. 7J shows a graph depicting the movement with the first bar closest to the y-axis representing WT, the second bar representing the JNCL mouse model, the third bar representing JNCL and PPARa null model.

[0059] FIG. 8A shows a graph depicting the MFI of TFEB with the first bar closest to the y-axis representing WT, the second bar representing the JNCL mouse model, the third bar representing JNCL model dosed with 8mg / kg / d, the fourth bar representing the JNCL and PPARa null model, and the fifth bar representing the JNCL and PPARa null model dosed with 8mg / kg / d.

[0060] FIG. 8B shows a graph depicting the MFI of SCMAS with the first bar closest to the y-axis representing WT, the second bar representing the JNCL mouse model, the third bar representing JNCL model dosed with 8mg / kg / d. the fourth bar representing the JNCL and PPARa null model, and the fifth bar representing the JNCL and PPARa null model dosed with 8mg / kg / d.

[0061] FIG. 9A shows a graph depicting the number of microglia per millimeter squared with the first bar closest to the y-axis representing WT, the second bar representing the JNCL mouse model, the third bar representing JNCL model dosed with 8mg / kg / d, the fourth bar representing the JNCL and PPARa null model, and the fifth bar representing the JNCL and PPARa null model dosed with 8mg / kg / d.

[0062] FIG. 9B shows a graph depicting the number of astrocytes per millimeter squared with the first bar closest to the y-axis representing WT, the second bar representing the JNCL mouse model, the third bar representing JNCL model dosed with 8mg / kg / d, the fourth bar representing the JNCL and PPARa null model, and the fifth bar representing the JNCL and PPARa null model dosed with 8mg / kg / d.

[0063] FIG. 9C shows a graph depicting the number of iNOS(+) cells per millimeter squared with the first bar closest to the y-axis representing WT, the second bar representing the JNCL mouse model, the third bar representing JNCL model dosed with 8mg / kg / d, the fourth bar representing the JNCL and PPARa null model, and the fifth bar representing the JNCL and PPARa null model dosed wi th 8mg / kg / d.

[0064] FIG. 10A shows a graph depicting the velocity measured in centimeters per seconds with the first bar closest to the y-axis representing WT, the second bar representing the JNCL and PPARa null model, and the third bar representing the JNCL and PPARa null model dosed with 8mg / kg / d, and the fourth bar representing the JNCL and PPARa null model dosed with vehicle.

[0065] FIG. 10B shows a graph depicting cumulative duration in seconds with the first bar closest to the y-axis representing WT, the second bar representing the JNCL and PPARa null model, and the third bar representing the JNCL and PPARa null model dosed with 8mg / kg / d, and the fourth bar representing the JNCL and PPARa null model dosed with vehicle.

[0066] FIG. 10C shows a graph depicting the distance traveled in centimeters with the first bar closest to the y-axis representing WT, the second bar representing the JNCL and PPARa null model, and the third bar representing the JNCL and PPARa null model dosed with 8mg / kg / d, and the fourth bar representing the JNCL and PPARa null model dosed with vehicle.

[0067] FIG. 10D shows a graph depicting the centerpoint moving with the first bar closest to the y-axis representing WT, the second bar representing the JNCL and PPARa null model, and the third bar representing the JNCL and PPARa null model dosed with 8mg / kg / d, and the fourth bar representing the JNCL and PPARa null model dosed with vehicle.

[0068] FIG. 10E shows a graph depicting the rototorod latency in seconds with the first bar closest to the y-axis representing WT, the second bar representing the JNCL and PPARanull model, and the third bar representing the JNCL and PPARa null model dosed with 8mg / kg / d, and the fourth bar representing the JNCL and PPARa null model dosed with vehicle.

[0069] FIG. 10F shows a graph depicting pole T-tum in seconds with the first bar closest to the y-axis representing WT, the second bar representing the JNCL and PPARa null model, and the third bar representing the JNCL and PPARa null model dosed with 8mg / kg / d, and the fourth bar representing the JNCL and PPARa null model dosed with vehicle.

[0070] FIG. 10G shows a graph depicting pole latency in seconds with the first bar closest to the y-axis representing WT, the second bar representing the JNCL and PPARa null model, and the third bar representing the JNCL and PPARa null model dosed with 8mg / kg / d, and the fourth bar representing the JNCL and PPARa null model dosed with vehicle.

[0071] DETAILED DESCRIPTION OF THE INVENTION

[0072] Throughout this disclosure, various quantities, such as amounts, sizes, dimensions, proportions and the like, are presented in a range format. It should be understood that the description of a quantity in range format is merely for convenience and brevity' and should not be construed as an inflexible limitation on the scope of any embodiment. Accordingly, the description of a range should be considered to have specifically disclosed all the possible subranges as well as all individual numerical values within that range unless the context clearly dictates otherwise. For example, description of a range such as from 1 to 6 should be considered to have specifically disclosed subranges such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4. from 2 to 6, from 3 to 6 etc., as well as individual values within that range, for example, 1.1, 2, 2.3, 4.62, 5, and 5.9. This applies regardless of the breadth of the range. The upper and lower limits of these intervening ranges may independently be included in the smaller ranges, and are also encompassed within the disclosure, subject to any specifically excluded limit in the stated range. Where the stated range includes one or both of the limits, ranges excluding either or both of those included limits are also included in the disclosure, unless the context clearly dictates otherwise.

[0073] The terminology' used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of any embodiment. As used herein, the singular forms “a,” “an” and "the" are intended to include the plural forms as well, unless the context clearly indicates otherwise. It will be further understood that the terms “includes,” “comprises,” “including” and / or “comprising,” when used in this specification, specify the presence of stated features, integers, steps, operations, elements, and / or components, but do not preclude the presence or addition of one or more other features, integers, steps, operations, elements, components, and / or groups thereof. As used herein, the term “and / or” includes anyand all combinations of one or more of the associated listed items. Additionally, it should be appreciated that items included in a list in the form of "at least one of A. B, and C” can mean (A); (B); (C); (A and B); (B and C); (A and C); or (A, B, and C). Similarly, items listed in the form of “at least one of A, B, or C” can mean (A); (B); (C); (A and B); (B and C); (A and C); or (A, B, and C).

[0074] Unless specifically stated or obvious from context, as used herein, the term “about’7in reference to a number or range of numbers is understood to mean the stated number and numbers + / - 10% thereof, or 10% below the lower listed limit and 10% above the higher listed limit for the values listed for a range.

[0075] The present disclosure provides for compositions and methods for treating Juvenile neuronal ceroid lipofuscinosis (JNCL) in a patient in need thereof. In other embodiments, are provided methods for treating or correcting a defective function of the lysosomal membrane glycoprotein Cln3. In any embodiment the methods comprise administering to a patient in need thereof an effective amount of an oral pharmaceutical composition comprising gemfibrozil. The disclosed methods contemplate that the pharmaceutical composition comprising gemfibrozil may be administered to the patient in any including one time per day, two times per day, and three times per day.

[0076] As used herein, the term “pharmaceutically acceptable carrier” means a nontoxic, inert solid, semi-solid or liquid filler, diluent, encapsulating material or formulation auxiliary of any type.

[0077] As used herein, the term “treatment” means to alleviate or reduce symptoms associated with a disease.

[0078] Some examples of materials which can serve as pharmaceutically acceptable carriers are sugars such as lactose, glucose and sucrose; starches such as com starch and potato starch; cellulose and its derivatives such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients such as cocoa butter and suppository waxes; oils such as peanut oil, cottonseed oil; safflower oil; sesame oil; olive oil; com oil and soybean oil; glycols such as propylene glycol; esters such as ethyl oleate and ethyl laurate; agar; buffering agents such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer's solution; ethyl alcohol, and phosphate buffer solutions, as well as other non-toxic compatible lubricants such as sodium lauryl sulfate and magnesium stearate, as well as coloring agents, releasing agents, coating agents, sweetening, flavoring and perfuming agents, preservatives and antioxidants can also be present in the composition, according to the judgment of the formulator. Other suitablepharmaceutically acceptable excipients are described in ‘‘Remington's Pharmaceutical Sciences;’ Mack Pub. Co., New Jersey, 1991, the contents of which are expressly incorporated herein by reference.

[0079] Methods of formulation are well known in the art (see, for example, Remington: The Science and Practice of Pharmacy, Mack Publishing Company, Easton, Pa., 19th Edition (1995)). Pharmaceutical compositions for use in accordance with the present disclosure can be in the form of sterile, non-pyrogenic liquid solutions or suspensions, coated capsules, lyophilized powders, or other forms known in the art.

[0080] Solid dosage forms for oral administration include, as illustrative but nonlimiting examples, capsules, tablets, pills, powders, thin films and granules. In solid dosage forms, the active compound may be mixed with at least one inert, pharmaceutically acceptable excipient or carrier. Illustrative, non-limiting examples of excipients or carriers include sodium citrate or dicalcium phosphate and / or a) one or more fillers or extenders (a filler or extender may be, but is not limited to, one or more selected from starches, lactose, sucrose, glucose, mannitol, and silicic acid), b) one or more binders (binders may be selected from, but not limited to. carboxymethylcellulose, alginates, gelatin, polyvinylpyrrohdinone, sucrose, and acacia), c) one or more humectants (a humectant may be, but is not limited to, glycerol), d) one or more disintegrating agents (disintegrating agents may be selected from, but are not limited to, agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, silicates, and sodium carbonate), e) one or more solution retarding agents (for example, but not limited to, paraffin), f) one or more absorption accelerators (selected from, but not limited to, quaternary ammonium compounds), g) one or more w etting agents (for example, but not limited to, acetyl alcohol and glycerol monostearate), h) one or more absorbents (selected from, but not limited to, kaolin and bentonite clay), and i) one or more lubricants (selected from, but not limited to. talc, calcium stearate, magnesium stearate, solid polyethylene glycols, and sodium lauryl sulfate). In the case of capsules, tablets and pills, for example, the dosage form may also comprise buffering agents.

[0081] Solid compositions of a similar type may also be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or milk sugar as well as high molecular weight polyethylene glycols and the like.

[0082] The solid dosage forms of tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells. Illustrative, non-limiting examples of coatings and shells include enteric coatings and other coatings / shells well known in the pharmaceutical formulating art. They may optionally contain opacifying agents and can also be of acomposition that they release the active ingredient(s) only, or preferentially, in a certain part of the intestinal tract, optionally, in a delayed manner. Examples of embedding compositions that may be used include, but are not limited to, polymeric substances and waxes.

[0083] The active compounds can also be in micro-encapsulated form with one or more excipients as noted above. The solid dosage forms of tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells. The coatings or shells may be, but are not limited to, enteric coatings, release-controlling coatings and other coatings in the pharmaceutical formulating art. In solid dosage forms, the active compound may be admixed with at least one inert diluent. The inert diluent may include, but is not limited to, one or more of, sucrose, lactose or starch. Dosage forms may also comprise additional substances other than inert diluents. The additional substances may be, but are not limited to, tableting lubricants and other tableting aids. The tableting lubricants and other aids may be, but are not limited to, magnesium stearate and microcrystalline cellulose. In the case of capsules, tablets and pills, for example, the dosage forms may also comprise buffering agents. They may comprise opacifying agents. They may be of a composition that releases the active ingredient(s) only, or preferentially, in a certain part of the intestinal tract. The release may be in a delayed manner. Examples of embedding compositions that can be used include, but are not limited to, polymeric substances and waxes.

[0084] Liquid dosage forms for oral administration include, but are not limited to, pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs. In addition to the active compounds, the liquid dosage forms may comprise one or more inert diluents. The inert diluents may be selected from those commonly used in the art. Illustrative, non-limiting examples of inert diluents include water or other solvents, solubilizing agents and emulsifiers (including, but not limited to, ethyl alcohol, isopropyl alcohol, ethyl carbonate, EtOAc, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oils (in particular, cottonseed, groundnut, com, germ, olive, castor, and sesame oils), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols and fatty7acid esters of sorbitan, and mixtures thereof). The oral compositions may comprise one or more adjuvants. Illustrative, non-limiting examples of adjuvants include wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

[0085] The liquid dosage forms may be in the form of pharmaceutical suspensions which are understood to be liquid dosage forms containing finely divided insoluble materials (the suspensoid) distributed somewhat uniformly throughout the suspending medium (suspending vehicle) in which the drug exhibits a minimum degree of solubility.

[0086] In still other embodiments, it is contemplated that the pharmaceutical composition may be part of a thin film administration form. See Karki et al.. (2016), "‘Thin films as an emerging platform for drug delivery,’’ Asian J Pharmaceutical Sci. 11 : pp. 559- 574. Generally, it is understood that thin films, alternatively referred as a thin and flexible layer of polymer with or without a plasticizer. Thin films provide means for targeting sensitive site that may not be possible with tablets or liquid formulations. Thin films have shown the capabilities to improve the onset of drug action, reduce the dose frequency and enhance the drug efficacy.

[0087] The amount of active ingredient, wherein the active ingredient is oral gemfibrozil that may be combined with the optional carrier materials to produce a single dosage form may vary depending upon the host treated and the particular mode of administration. The specific dose level for any particular patient may depend upon a variety of factors including the activity of the specific compound employed, the age, body weight, general health, sex, diet, time of administration, route of administration, rate of excretion, drug combination, and the severity of the particular disorder or disease undergoing therapy. A therapeutically effective amount for a given situation can be readily determined by routine experimentation and is within the skill and judgment of the ordinary clinician.

[0088] In accordance with certain methods of treatment disclosed in the present application, progression of various disorders is slowed or stopped in a patient (a patient may be a human, a lower mammal, or a warm blooded animal), by administering to the patient an effective amount of the i) oral gemfibrozil and / or gemfibrozil and vitamin A; or ii) cinnamic acid, in such amounts, and for such time as is necessary', to achieve the desired result. An amount of a compound that is effective to slow' or stop the progression of a disease or disorder may refer to a sufficient amount of the compound to treat the disease or disorder at a reasonable benefit / risk ratio applicable to any medical treatment.

[0089] The total daily usage of the compounds and compositions of the present disclosure may be decided by the attending physician within the scope of sound medical judgment. The specific therapeutically effective dose level for any particular patient may depend upon a variety of factors including the disease or disorder being treated and the severity of the disorder; the activity of the specific compound employed; the specific composition employed; the age, body weight, general health, sex and diet of the patient; the time of administration, route of administration, and rate of excretion of the specific compound employed; the duration of the treatment; and drugs used in combination or coincidental with the specific compound employed.

[0090] The “effective amount'’ or dose of a compound of the present disclosure, such as i) gemfibrozil, to be administered to warm-blooded animals (e.g.. humans) may vary depending upon the disorder to be treated, the method or mode of drug administration and the desire to minimize any known side effects. In connection with certain neurodegenerative disorders, such as, juvenile Batten disease, the effective amount may be from about 0.01 mg / kg to about 5.0 g / kg per day, or any amount or sub-range thereof. In preferred embodiments, the dosage is in the range of about 0.01 mg / kg / day to about 200 mg / kg / day.

[0091] The administration may be once per day, twice per day, or more than tw o times per day. Additionally, in some embodiments, a patient may receive the active ingredients by multiple administration methods including combinations of oral and buccal or sublingual administration. The present disclosure encompasses any combination of the administration techniques described or contemplated herein.

[0092] In some embodiments, the composition comprising gemfibrozil is provided to a child ranging in age from about 6 months to about 10 years old in an effort to reduce or prevent symptoms from forming before onset of JNCL. In some embodiments, the composition is administered to a child of at least about 6 months, at least about 1 year, at least about 1.5 years, at least about 2 years, at least about 2.5 years, at least about 3 years, at least about 3.5 years, at least about 4 years, at least about 4.5 years, at least about 5 years, at least about 5.5 years, at least about 6 years, at least about 6.5 years, at least about 7 years, at least about 7.5 years, at least about 8 years, at least about 8.5 years, at least about 9 years, at least about 9.5 years, or at least about 10 years of age.

[0093] In some embodiments, the composition comprising gemfibrozil is administered continuously for the life of the human patient.

[0094] In some embodiments, the method of treatment further comprises coadministering the composition comprising gemfibrozil with a second therapy. The second therapy may be a gene therapy, an oral medication, a dietary regimen, or the like.

[0095] In addition the aspects and embodiments described and provided elsewhere in the present disclosure, the following non-limiting list of embodiments are also contemplated.

[0096] 1. A method for treating a patient diagnosed with or suspected of having juvenile neuronal ceroid lipofuscinosis (JNCL) in need thereof, comprising administering to the patient a composition comprising a therapeutically effective amount of gemfibrozil, or a pharmaceutically acceptable salt thereof, wherein the administering results in a reduction in symptoms associated with JNCL, thereby treating the patient.

[0097] 2. The method of clause 1, wherein the patient is at least about 6 months old.

[0098] 3. The method of clauses 1 or 2, wherein the patient is at least about 3 years old.

[0099] 4. The method of any one of clauses 1-3, wherein the patient is between about6 months to about 10 years old.

[0100] 5. The method of any one of clauses 1-4, wherein the composition further comprises at least one pharmaceutically acceptable excipient or carrier.

[0101] 6. The method of any one of clauses 1-5, wherein the composition is formulated to be administered orally to the patient.

[0102] 7. The method of any one of clauses 1-6, wherein the patient is not displaying symptoms of JNCL before administering the composition.

[0103] 8. The method of any one of clauses 1-7, wherein the composition is administered at least once daily.

[0104] 9. The method of any one of clauses 1-8, wherein the composition is administered at least twice daily.

[0105] 10. The method of any one of clauses 1-9 further comprising co-administering a second therapy.

[0106] 11. The method of clause 10. wherein the second therapy is a gene therapy.

[0107] 12. A composition comprising gemfibrozil, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for treating JNCL in a patient.

[0108] 13. The composition of clause 12, wherein the composition is formulated to be given orally to the patient.

[0109] 14. The composition of clauses 12 or 13, wherein the patient is between the ages of about 3 to about 10 years old.

[0110] 15. The composition of clauses 12-14, wherein the patient is at least about 6 months old.

[0111] 16. A composition comprising gemfibrozil, or a pharmaceutically acceptable salt thereof, for us in a therapy to treat JNCL in a patient.

[0112] 17. The composition of clause 16, wherein the composition is formulated to be given orally to the patient.

[0113] 18. The composition of clauses 16 or 17, wherein the patient is between the ages of about 3 to about 10 years old.

[0114] 19. The composition of any one of clauses 16-18, wherein the patient is at least about 6 months old.

[0115] 20. A method for treating or correcting a defective lysosomal transmembrane glycoprotein Cln3 in a patient in need thereof, comprising administering to a patient in needthereof an effective amount of a pharmaceutical composition comprising gemfibrozil and at least one pharmaceutically acceptable excipient or carrier.

[0116] Materials and Methods

[0117] Reagents: Different molecular biology-grade chemicals were obtained from Sigma-Aldrich. Gemfibrozil and methyl cellulose were purchased from Spectrum Chemicals. Primary’ antibodies, their sources, and the concentrations used are listed in Table 1. Alexa Fluor antibodies used in immunostaining were purchased from Jackson ImmunoResearch Laboratories. IR-dye-labeled reagents used for immunoblotting were received from Li-Cor Biosciences.

[0118] Animals and Gemfibrozil Treatment: Homozygous CLN3 (Cln3Aex7 / 8; Jackson Laboratory) mice were used as a model of JNCL. These mice were referred to as the Cln3AJNCL through the rest of the manuscript. Cln3AJNCL mice were screened by genotyping that was performed by PCR on DNA obtained from tail biopsy samples using following primers:

[0119] Common: 5 -CACTTGGGAGATTGTGAATTTG-3’ SEQ. ID. NO: 1

[0120] Mutant reverse: 5 -GGTGCTCCCAGCCTCTAGGT-3’ SEQ. ID. NO:2

[0121] Wild type reverse: 5’-GAGATAGGGTTTTGCTGTGC-3’ SEQ. ID. NO:3

[0122] Wild type (WT) mice from the same background were used as controls.Cln3AJNCL mice were crossed with PPARa null mice to create Cln3AJNCLAPPAR“ bi-genic mice. Animals were maintained and experiments were conducted in accordance with National Institutes of Health guidelines and were approved by the Rush University Medical Center Institutional Animal Care and Use Committee. Three-month-old Cln3AJNCL and Cln3AJNCLAPPARramice of both sexes were treated with different dose of gemfibrozil (4 and 8 mg / kg body wt / d) solubilized in 100 pl 0. 1% methyl cellulose (MC) via gavage for 3 months followed by monitoring locomotor activity and biochemical assays. Another group of Cln3AJNCL mice also received only MC as vehicle.

[0123] Table 1. Antibodies, sources, applications, and dilutions used

[0124] WB, western blot; IF, immunofluorescence; GFAP, glial fibrillary acidic protein; Iba-1, ionized calcium-binding adaptor molecule 1; ChIP, chromatin immunoprecipitation; IL-ip, interleukin IP; TNFa, tumor necrosis factor a; PPARa, peroxisome, proliferator-activated receptor a; TFEB, transcription factor EB

[0125] Immunohistochemistry (IHC)

[0126] For immunohistochemistry, mice were anesthetized and intracardially perfused with IX PBS followed by 4% paraformaldehyde (PF A) in 0. 1 M phosphate buffer, pH 7.4. The brains were post fixed in PFA overnight at 4 °C and were then transferred to phosphate buffer containing 30% sucrose at 4 °C. Somatosensory barrel cortex sections were cut and saved in serial order at - 20 °C until immunostained. For this, the hemi brains incubated in 30% sucrose were washed thoroughly in PBS cryo-sectioned using a sliding microtome (American opticals 860). Prior to staining, 40pM free floating somatosensory barrel field cortex sections were washed thoroughly in PBS. The sections were blocked using 2% BSA in PBSTT (PBS + Triton X-100 + Tween-20) for 1 h. Next, the sections were incubated with primary antibody in 1% PBSTT at 4 °C overnight. The following day, sections were washed in PBSTT and incubated with 488 or 647- conjugated secondary' antibody (Jackson ImmunoResearch Laboratories) for 3 h at room temperature. Following washes in PBSTT, the sections were mounted on glass slides (Patel et al., 2019; Raha et al., 2020). The samples were visualized under OlympusBX41fluorescence microscope equipped with a Hamamatsu ORCA-03G camera. For DAB staining, somatosensory motor cortex sections were stained for GFAP and Ibal using the Vectastain DAB protocol, mounted and observed under an Olympus bright field microscope. Optical Density measures were produced using ImageJ software (Raha et al., 2020; Paidi et al., 2021b) as described before (Varghese et al., 2014). Counting analysis was performed using the Olympus Microsuite V software (Waltham, MA, USA) for imaging applications with the help of touch counting module (Corbett et al., 2015).

[0127] Immunoblotting

[0128] Western blotting was conducted as described earlier (Jana et al., 2012; Rangasamy et al., 2018; Chandra and Pahan, 2019). After 12 weeks of treatment, mice were perfused with PBS and from half part of the mice brain we collected somatosensory motor neuron region, homogenized in RIPA buffer. The supernatant was collected and analyzed for protein concentration via the Bradford method (Bio-Rad). SDS sample buffer was added to protein samples and boiled for 5 min. Denatured samples were electrophoresed on 10 or 12% Bis-Tris SDS polyacrylamide gels in a continuous buffer system, transferred onto a nitrocellulose membrane (Bio-Rad) using the Thermo-Pierce Fast Semi-Dry Blotter. The membrane was then washed for 1 min in TBS plus Tween 20 (TBST) and blocked for 1 h in TBST containing BSA. Next, membranes were incubated at 4°C under shaking conditions with primary antibodies followed by washing of membranes in TBST for 1 h. Membranes were then incubated in secondary antibodies for 1 h at room temperature, washed for one more hour, and visualized under the Odyssey® Infrared Imaging System (Li-COR, Lincoln, NE, USA). Blots were converted to binary', analyzed using ImageJ (NIH), and normalized to the -actin loading control.

[0129] In situ chromatin immunoprecipitation (ChIP) assay

[0130] In situ ChIP was performed as described. Briefly, animals were perfused with PBS and then PBS containing 4% paraformaldehyde followed by isolation of somatosensory barrel field (SBF) cortex for the isolation of DNA using the phenol-chloroform-isopropyl alcohol method of DNA isolation. ChIP was performed on the cell lysate by overnight incubation at 4 °C with 2 pg of anti-PPARa, anti-CBP or anti-RNA polymerase II antibodies followed by incubation with protein G agarose (Santa Cruz Biotechnology) for 2 h. The beads were then washed with cold IP buffer, and a total of 100 pl of 10% Chelex (10 g / 100 ml H2O) was added to the washed protein G beads and vortexed. The Chelex / protein G bead suspension was boiled for 10 min and then allowed to return to room temperature. Proteinase K (100 pg / ml) was then added, and the beads were incubated for 30 min at 55°C while shaking.followed by another round of boiling for 10 min. The suspension was centrifuged, and the supernatant was collected. This elute was used for conducting semi-quantitative and real-time PCR. The PPRE-containing fragment of the mouse Tfeb promoter was amplified using the following primers: sense: 5'-GAA CAT TCC AGG TGG AGG CA-3', (SEQ. ID. NO: 5) antisense: 5’-CCC CCA AC A CAT GCT TCT CT-3' (SEQ. ID. NO: 6). For real-time PCR, data were normalized with the input and the fold change with respect to the untreated control was calculated.

[0131] Behavioral Analyses:

[0132] Open Field Test: It was performed as described earlier (Patel et al., 2018; Raha et al., 2020; Paidi et al., 2021b). Briefly, each mouse was allowed to freely explore an open field arena for 5 min. The testing apparatus was a classic open field (i.e.. a wooden floor square arena, 40 x 40 cm, with walls 30 cm high). A video camera (Basler Gen I Cam-Basler acA 1300-60) connected to aNoldus computer system was placed above the box. Each mouse was placed individually on the center of the arena and the locomotor activity7and other parameters like velocity, total distance travelled, and center time frequency was monitored for 5 minutes using live video tracking system (Noldus System). The central area was arbitrarily defined as a square of 20 x 20 cm (half the total area).

[0133] Rotarod Test: The rotarod uses a motor-driven, rotating rod to measure limb motor coordination and balance of mice. It was measured on a rotarod apparatus (ENV-576M; Med-associates Inc.), using protocol described earlier (Chandra and Pahan, 2019; Patel et al., 2019; Raha et al., 2020). Briefly, mice were transported (within their home cage) to acclimate to the testing room for 1 h prior to trial. Before acquisition, the parameters of the Rotarod system equipped with automatic fall detector such as start speed and acceleration were carefully checked before acquisition. Each mouse was placed on the confined section of the rod and trial was initiated with a smooth increase in speed from 4 rpm to 40 rpm for 5 min. If the mouse did not fall from the rod, it was removed from the rod after 5 min.

[0134] Pole Test: In order to test motor coordination and spatial awareness, a vertical test was performed as described before (Chandra et al., 2016; Raha et al., 2020). A vertical wooden pole with a rough surface (50 cm in height and 1 cm in diameter) was placed in the home cage. Mice were acclimatized to the pole over three trials of 120 s each. Each trial was separated by 60s, and during behavioral testing, each mouse was tested thrice.

[0135] Detection of storage materials: It was performed by monitoring subunit C of mitochondrial ATP synthase (SCAMS) by immunofluorescence as described before (Ghosh et al., 2017). Please see Table 1 for details on antibody dilutions. DAPI was used to monitornucleus. SCAMS associated fluorescence intensity was quantified by using the Olympus Microsuite V Software. Briefly, captured images were opened in the infinity image viewer and the contour was drawn around the granules to obtain the fluorescence intensity.

[0136] Densitometric analysis: Protein blots were analysed using Imaged (NIH, Bethesda, MD) and bands were normalized to their respective P-actin loading controls. Data are representative of the average fold change with respect to control for three independent experiments.

[0137] Statistical analysis: Statistical analyses were performed using GraphPad Prism 8.0 (GraphPad Software. Inc., La Jolla, CA). Mouse behavioral parameters were examined by an independent one-way ANOVA using SPSS. Homogeneity of variance between test groups was examined using Levene’s test. Post-hoc analyses were conducted using Tukey’s tests. Other data were expressed as means ± SD of three independent experiments. Statistical differences between means were calculated by the Student's t-test (two-tailed). A p- value of less than 0.05 (p <0.05) was considered statistically significant.

[0138] The following examples are intended to illustrate some embodiments of the present disclosure and are not intended to limit the disclosure or scope of the claims in any manner.

[0139] EXAMPLES

[0140] EXAMPLE 1: Oral administration of gemfibrozil attenuates glial activation in somatosensory barrel field (SBF) cortex of Cln3AJNCL mice

[0141] Since gemfibrozil inhibits glial activation and associated inflammation, we investigated whether gemfibrozil treatment was capable of suppressing astroglial activation in an animal model of JNCL. Although multiple mouse models of JNCL have been generated affecting the orthologous murine Cln3 gene to mimic the human mutation, Cln3 Aex7 / 8 knock- in mice leading to 1.02 kb genomic deletion in the Cln3 eliminates exons 7 and 8 surrounding intronic DNA corresponds to the most common deletion found in JNCL patients. Accordingly, a Cln3Aex7 / 8 knock-in (Cln3AJNCL) mice, an accepted JNCL model, was selected to study the efficacy of gemfibrozil. Turning to FIGs. 1 A to ID show the effect of gemfibrozil on glial activation in the somatosensory barrel field (SBF) cortex of the Cln3AJNCL mice. Briefly, the graphs show the results of three-month Cln3AJNCL old mice (n=6 per group) treated with different doses of gemfibrozil (4 or 8 mg / kg body wt) daily via gavage for 3 months followed by monitoring the protein level of different proinflammatory molecules by Western blot (not shown). Bands from the western blot were scanned and values (cytokine / actin) presented asrelative to WT-control were rendered in a bar graph shown in FIG. 1A, GFAP and FIG. IB, Ibal. DAB staining of SBF cortical sections for GFAP (not shown) and Ibal (not shown) were used to perform cell counts. The results of the cell counts from a defined brain section were rendered in a bar graph with FIG. 1 C showing the cell count GFAP-positive cells and FIG. ID showing Iba-1 -positive cells. The cell count was performed in one section (two images per section) of each of six different mice (n=6) per group. ** p < 0.01; *** p < 0.001.

[0142] As expected, increased protein level of astroglial marker GFAP was seen in somatosensory barrel field (SBF) cortex of 6-month-old Cln3AJNCL mice as compared to age- matched WT mice (FIG. 1 A). However, oral administration of different doses (4 and 8 mg / kg body weight / day) of gemfibrozil strikingly reduced the level of GFAP protein as shown in FIG. 1A. Immunohistochemical analysis of SBF cortex of Cln3AJNCL mice also exhibited morphological characteristic of reactive astrogliosis with intense GFAP immunoreactivity (not shown). The number of activated astrocytes / mm2was also significantly higher in Cln3AJNCL mice compared to WT mice (FIG. 1C). However, gemfibrozil treatment markedly inhibited astrogliosis in the SBF cortex of Cln3AJNCL mice (FIG. 1C).

[0143] Similarly, the gemfibrozil treatment all had an effect on microgliosis in Cln3AJNCL mice. Reactive microglia are thought to be key contributors in several neurodegenerative disorders including JNC with some studies showing evidence of early microglial activation may predict regions where neuronal loss occurs later in the disease process of a JNCL mouse model. A western blot analysis showed that the Ibal protein level was markedly enhanced in the SBF cortex of Cln3AJNCL mice compared to WT mice and that gemfibrozil treatment normalized the level of Ibal in Cln3AJNCL mice (FIG. IB). To confirm the finding, we also performed immunohistochemistry with antibodies against Ibal and found normal-appearing microglia in WT mice, but reactive ones in Cln3AJNCL mice (FIG. ID). However, oral gemfibrozil treatment inhibited microglial activation and decreased the number of activated microglia in the SBF cortex of Cln3AJNCL mice (FIG. ID).

[0144] Oral gemfibrozil suppresses pro-in flammat ory molecules in the SBF cortex of Cln3AJNCL mice. Several reports indicate that proinflammatory gene expression is increased in the SBF cortex of Cln3AJNCL mice compared to WT mice. Turning to FIGs. 2A-2D show the effect of gemfibrozil on the level of different proinflammatory molecules in the SBF cortex of Cln3AJNCL mice. Three-month old Cln3AJNCL mice (n=6 per group) were treated with different doses of gemfibrozil (4 and 8 mg / kg body wt) daily via gavage for 3 months followed by monitoring the protein level of different proinflammatory molecules by Western blot (not shown). Bands from the western blot were scanned and values (cytokine / actin) presented asrelative to WT-control (FIG. 2A, iNOS; FIG. 2B, Pro-IL-1 P; FIG. 2C. IL-1P; FIG. 2D, TNFa). Immunostaining of SBF cortical sections for GFAP (not shown) and Ibal (not show n). Results represent analysis of one section of each of six different mice per group. Results are mean+SEM of six mice per group. * p < 0.05; ** p < 0.01; *** p < 0.001. Since activated glial cells produce different proinfl ammatory molecules and gemfibrozil treatment inhibited glial activation, we investigated the status of different pro-inflammatory molecules in gemfibrozil- treated and untreated Cln3AJNCL mice. Western blot analysis of SBF cortical tissues showed significant increase in protein levels of iNOS, pro-IL-ip, IL- ip, and TNFa in Cln3AJNCL mice as compared to wild type mice (FIGs. 2A to 2D). However, consistent to the inhibition of gliosis, gemfibrozil treatment decreased the levels of iNOS, pro-IL-ip, IL-ip, and TNFa in the SBF cortex of Cln3AJNCL mice (FIGs. 2A to 2D). This effect was more prominent at the higher dose (8 mg / kg body weight / d) of gemfibrozil (FIGs. 2A to 2D). Moreover, the expression of nitrosative stress marker inducible nitric oxide synthase (iNOS) was upregulated in GFAP-positive astrocytes and Ibal -positive microglia in SBF cortex of Cln3AJNCL mice, which was strongly inhibited by gemfibrozil treatment (not shown).

[0145] EXAMPLE 2:

[0146] Oral gemfibrozil upregulates TFEB and lowers the burden of storage material in SBF cortex of Cln3AJNCL mice

[0147] Accumulation of autofluorescent storage material and activation of glia are early neuropathological hallmarks of CLN3 batten disease that are reciprocated in Cln3AJNCL mice. Several studies have demonstrated that transcription factor-EB (TFEB), a master regulator of lysosomal biogenesis, plays a critical role in cellular clearance in neurodegenerative storage diseases. Turning to FIGs. 3A to 3D show the effect of gemfibrozil on the level of TFEB and storage materials in the SBF cortex of Cln3AJNCL mice. Three- month old Cln3AJNCL mice (n=6 per group) were treated with different doses of gemfibrozil (4 and 8 mg / kg body wt) daily via gavage for 3 months followed by double-labeling of SBF cortical sections with antibodies against TFEB and NeuN (not shown). Mean fluorescence intensity (MFI) of TFEB (FIG. 3A) and NeuN (FIG. 3B) was quantified in two different sections (two images per section) of each of six different mice (n=6) per group using NIH Image J software. SBF cortical homogenates were immunoblotted for TFEB. Actin was run as a loading control (not shown). FIG. 3C show s bands from the immunoblot w ere scanned and values (TFEB / actin) presented as relative to WT-control. Results are mean + SEM of six mice per group. Storage pigments were observed in SBF cortical sections by immunofluorescence analysis of subunit c of mitochondrial ATP synthase (SCMAS). DAPI was used to visualizenucleus, (not shown) MFI of SCMAS was quantified in one section (two images per section) of each of six different mice (n=6) per group using NIH Image J software. (FIG. 3D) *** p < 0.001. Double-label immunofluorescence analysis of SBF cortical sections for TFEB and NeuN revealed marked decrease in both TFEB (FIG. 3A) and NeuN (FIG. 3B) in 6-month old Cln3AJNCL mice as compared to age-matched WT mice. On the other hand, gemfibrozil treatment increased and / or normalized the level of both TFEB (FIG. 3A) and NeuN (FIG. 3B) in the SBF cortex of Cln3AJNCL mice. Western blot of SBF cortical tissues for TFEB also corroborates this finding (FIG. 3C).

[0148] TFEB is a transcription factor and its target genes are ultimately involved in lysosomal biogenesis and autophagy. Therefore, to confirm the activation of TFEB, we monitored the status of tripeptidylpeptidase 1 (TPP1) and p62. molecules that are controlled by TFEB, in the SBF cortex of Cln3AJNCL mice. Turning to FIGs. 4A to 4D show the effect of gemfibrozil on the autophagy pathway in the SBF cortex of Cln3AJNCL mice. Three-month old Cln3AJNCL mice (n=6 per group) were treated with gemfibrozil (8 mg / kg body wt) dailyvia gavage for 3 months followed by monitoring the level of tripeptidylpeptidase 1 (TPP1) and p62 by Western blot (not shown). Actin was run as a loading control. Bands from the western blot were scanned and values (TPP1 / actin, FIG. 4A; p62 / Actin, FIG. 4B) presented as relative to WT-control. Results are mean + SEM of four mice per group. SBF cortical sections were double-labeled with antibodies against NeuN & TPP1 (not shown) and NeuN & p62 (not shown). MFI of TPP1 (FIG. 4C) and p62 (FIG. 4D) was quantified in two different sections (two images per section) of each of six different mice (n=6) per group using NIH Image J software. *** p < 0.001; NS, not significant. As evident from Western blot analysis, gemfibrozil treatment markedly increased the level of both TPP1 (FIG. 4 A) and p62 (FIG. 4B) in the SBF cortex of Cln3AJNCL mice. To confirm this finding further, we performed doublelabel immunofluorescence analysis that also exhibited marked increase in both TPP1 (FIG. 4C) and p62 (FIG. 4D) in the SBF cortex of Cln3AJNCL mice upon gemfibrozil treatment. These results suggest that gemfibrozil treatment is capable of stimulating the activated form of TFEB and that gemfibrozil increases autophagy in the CNS of Cln3AJNCL mice.

[0149] Next, we investigated whether gemfibrozil treatment could reduce the level of autofluorescent storage material containing SCMAS in SBF cortex of Cln3AJNCL mice. Expectedly, w e found striking accumulation of SCMAS in the SBF cortex of Cln3AJNCL mice as compared to WT mice (FIG. 3D). Yet consistent to the upregulation of TFEB, gemfibrozil treatment steered to a striking decrease in SCMAS in the SBF cortex of Cln3AJNCL mice (FIG. 3D).

[0150] EXAMPLE 3:

[0151] Oral gemfibrozil improves locomotor activities in Cln3AJNCL mice: Decreasing functional impairment is definitely a therapeutic goal of neuroprotection for JNCL patients. Similar to that observed in JNCL patients, Cln3AJNCL mice also exhibit motor deficits. Therefore, locomotor activities were checked in gemfibrozil-treated and untreated 6- month old Cln3AJNCL mice. Turning to FIGs. 5A to 5F show the effect of gemfibrozil on locomotor activities of Cln3AJNCL mice. Three-month old Cln3AJNCL mice (n=6 per group) were treated with different doses of gemfibrozil (4 and 8 mg / kg body wt) daily via gavage. After 3 months of treatment, open field behavior was monitored by Noldus tracking software to visualized as a heat map (not shown); FIG. 5A, velocity; FIG. 5B, cumulative distance; FIG. 5C. distance moved; FIG. 5D, center point moving; FIG. 5E, pole latency; FIG. 5F, rotarod latency. Results are mean + SEM of six mice per group. ** p < 0.01 ; *** p < 0.001. As expected, Cln3AJNCL mice displayed decrease in horizontal activity (not shown), velocity (FIG. 5A), cumulative duration (Fig. 5B), total distance traveled (Fig. 5C), and center point moving (Fig. 5D) as compared to age-matched WT mice. In pole test. WT mice climbed down the pole quickly and touched the base of the pole without hesitation, whereas Cln3 AJNCL mice showed abnormal behavior, which included turning upward, falling of the pole, slowly descending, freezing on the pole, and hesitating to touch the base of the pole (FIG. 5E). Similarly as evident from rotorod latency (FIG. 5F), Cln3 JNCL mice performed very poorly on rotorod. affirming the compromised motor coordination and muscle strength experience by these animals. However, oral administration of gemfibrozil significantly improved open-field, pole and rotorod activities of Cln3 AJNCL mice (FIGs. 5A to 5F), showing improved locomotor performance of Cln3 JNCL mice by gemfibrozil. Therefore, it can be surmised that reduction of batten pathology positively correlates with improved motor functions in gemfibrozil-treated Cln3AJNCL mice.

[0152] EXAMPLE 4:

[0153] Oral administration of gemfibrozil stimulates the recruitment of PPARa to the TFEB gene promoter in SBF cortex of Cln3AJNCL mice

[0154] Next, we investigated mechanism by which gemfibrozil upregulated TFEB to prevent the accumulation of storage materials in Cln3 AJNCL mice. Since gemfibrozil is know n to activate peroxisome proliferator-activated receptor a (PPARa), we examined the role of PPARa. Turning to FIGs. 6A to 6D show the oral administration of gemfibrozil increases the level of PPARa and the recruitment of PPARa to Tfeb promoter in the SBF cortex of Cln3AJNCL mice. Three-month old Cln3AJNCL mice (n=6 per group) were treated withgemfibrozil (8 mg / kg body wt) daily via gavage for 3 months followed by monitoring the protein level of PPARa by Western blot (not shown). Bands from the western blot were scanned and values (PPARa / actin) presented as relative to WT-control (FIG. 6A). Immunostaining of SBF cortical sections for PPARa and NeuN (not shown). Mean fluorescence intensity (MFI) of PPARa was quantified in two different sections (two images per section) of each of six different mice (n=6) per group using NIH Image J software (FIG. 6B). Schematic representation of the mouse Tfeb gene promoter with PPRE (SEQ. ID. NO. 4) (FIG. 6C). In situ ChIP for PPARa, CBP and RNA-Polymerase followed by semi-quantitative (not shown) and quantitative PCR (FIG. 6D) analyses were performed on the SBF cortex. *** p < 0.001. Western blot results showed that the protein level of PPARa markedly decreased in the SBF cortex of Cln3AJNCL mice as compared to wild type mice (FIG. 6A and 6B). On the other hand, gemfibrozil treatment significantly increased and / or restored PPARa level in the CNS of Cln3AJNCL mice (FIG. 6A and 6B). Double-label immunofluorescence of cortical sections of gemfibrozil-treated and untreated Cln3AJNCL mice for NeuN and PPARa also validates this finding (FIG. 6D).

[0155] We have demonstrated that activation of PPARa transcriptionally upregulates Tfeb and stimulates lysosomal biogenesis in brain cells. Therefore, by employing in-situ ChIP we investigated whether oral gemfibrozil treatment stimulated the recruitment of PPARa to the Tfeb gene promoter in vivo in the SBF cortex of Cln3AJNCL mice. FIG. 6C indicates the presence of a consensus PPRE (SEQ ID NO:4) in Tfeb gene promoter. After immunoprecipitation of chromatin fragments by antibodies against PPARa, we were able to amplily a 200-bp fragment encompassing the PPRE of the Tfeb promoter in SBF cortex of WT, but not Cln3AJNCL, mice (FIG. 6D), indicating decreased enrolment of PPARa to the Tfeb gene promoter in the CNS of Cln3AJNCL mice. However, gemfibrozil treatment markedly restored / increased the recruitment of PPARa to the Tfeb gene promoter in the SBF cortex of Cln3AJNCL mice (FIG. 6D). Similarly, we also observed decreased employment of CREB- binding protein (CBP), an important histone acetyl transferase, and RNA polymerase in the Tfeb promoter in the CNS of ACln3 mice, which was restored by gemfibrozil treatment (FIG. 6D). These results are specific as no product amplification was observed in immunoprecipitants with control IgG. Together, these results indicate that oral gemfibrozil stimulates the recruitment of PPARa to the Tfeb promoter in vivo in the CNS of Cln3AJNCL mice.

[0156] EXAMPLE 5:

[0157] Knockdown of PPARa aggravates JNCL pathologies and worsens locomotor performance in Cln3AJNCL mice

[0158] Turning to FIGs. 7A to 7J show the deletion of PPARa aggravates the disease process in Cln3AJNCL mice. Genetic screening of Cln3AJNCL mice lacking PPARa (Cln3AJNCLAPPARa) (not shown). Three-month old Cln3AJNCL and Cln3AJNCLAPPARa mice (n=6 per group) were treated with gemfibrozil (8 mg / kg body wt) daily via gavage for 3 months followed by monitoring the protein level of Iba-1 and GFAP by Western blot (not shown). Bands from the western blot were scanned and values (cytokine / actin) presented as relative to WT-control (FIG. 7 A; Ibal; FIG. 7B, GFAP). Immunostaining of SBF cortical sections for SCMAS (not shown). MFI of SCMAS was quantified in two different sections (two images per section) of each of six different mice (n=6) per group using NIH Image J software. (FIG. 7C) Mice were monitored for pole test (FIG. 7D, pole T-tum; FIG. 7E. pole latency), rotorod test (FIG. 7F) and open field behavior (FIG. 7G, velocity; FIG. 7H, cumulative duration; FIG. 71, Distance travelled; FIG. 7J, movement). Results are mean + SEM of six mice per group. * p < 0.05; ** p < 0.01; *** p < 0.001. To further reveal the role of PPARa in JNCL pathologies, we crossed Cln3AJNCL mice with PPARa- / - mice to generate Cln3AJNCLAPPARa mice (not shown). Interestingly, deletion of PPARa from Cln3AJNCL mice increased glial activation in the SBF cortex as evident from upregulation of Iba-1 and GFAP (FIG. 7A and FIG. 7B). Immunofluorescence analysis also demonstrated increased accumulation of SCMAS in the SBF cortex of Cln3AJNCLAPPARa mice as compared to Cln3AJNCL mice (FIG. 7C).

[0159] Next, we compared locomotor activities between Cln3AJNCL and Cln3AJNCLAPPARa mice. As evident from pole test, Cln3AJNCLAPPARa mice took longer time to make T turn (FIG. 7D) and descend the vertical pole (FIG. 7E) as compared to Cln3AJNCL mice. Similarly, Cln3AJNCLAPPARa mice exhibited poor performance on rotorod as compared to Cln3AJNCL mice (FIG. 7F). Consistently, Cln3AJNCL mice were more efficient than Cln3AJNCLAPPARa mice in overall locomotor activities including velocity (FIG. 7G), cumulative duration (FIG. 7H), distance travelled (FIG. 71), and movement (FIG. 7J). These results suggest that PPARa may play an important role in JNCL.

[0160] EXAMPLE 6:

[0161] Oral gemfibrozil upregulates TFEB. decreases storage materials and reduces glial activation in the SBF cortex of Cln3AJNCL mice via PPARa

[0162] To confirm that gemfibrozil does in fact require PPARa to exert its neuroprotective effects in Cln3AJNCL mice, we monitored the level of TFEB and the buildup of storage material SCMAS in SBF cortex of gemfibrozil-treated Cln3AJNCL mice andCln3AJNCLAPPARa mice. Turning to FIGs. 8A and 8B show the effect of gemfibrozil on the level of TFEB and storage materials in the SBF cortex of Cln3AJNCL and Cln3AJNCLAPPARa mice. Three-month old Cln3AJNCL and Cln3AJNCLAPPARa mice (n=6 per group) were treated with gemfibrozil (8 mg / kg body wt) daily via gavage for 3 months followed by double-labeling of SBF cortical sections for NeuN and TFEB (not shown). MFI of TFEB was quantified in two different sections (two images per section) of each of six different mice (n=6) per group using NIH Image J software (FIG. 8A). SBF cortical sections were also immunostained with antibodies against SCMAS (not shown). DAPI was used to visualize nucleus. MFI of SCMAS was quantified in two different sections (two images per section) of each of six different mice (n=6) per group using NIH Image J softw are (FIG. 8B). Results are mean + SEM of six mice per group. *** p < 0.001; NS. not significant. In contrast to the upregulation of TFEB and the reduction of storage materials in Cln3AJNCL mice, gemfibrozil treatment remained unable to increase TFEB (FIG. 8A) and decrease storage materials (FIG. 8B) in Cln3AJNCLAPPARa mice. Neuronal loss is seen in Cln3AJNCL mice and although gemfibrozil protected NeuN-positive neurons in Cln3AJNCL mice (FIG. 3B), such neuronal protection was not seen by gemfibrozil in Cln3AJNCLAPPARtmice (not shown). These results suggest that gemfibrozil involves PPARa for the upregulation of TFEB, reduction of storage materials and the protection of neurons in the CNS of Cln3AJNCL mice.

[0163] Next, we examined whether gemfibrozil entails PPARa to reduce glial activation in the CNS of Cln3AJNCL mice. Turning to FIGs. 9A to 9C show the effect of gemfibrozil on glial activation in the SBF cortex of Cln3AJNCL and Cln3AJNCLAPPARa mice. Three-month old Cln3AJNCL and Cln3AJNCLAPPARa mice (n=6 per group) were treated with gemfibrozil (8 mg / kg body wt) daily via gavage. After 3 months of treatment, mice were perfused and SBF cortical sections were double-labeled for Ibal & iNOS (not shown) and GFAP & iNOS (not shown). DAPI was used to visualize nucleus. Ibal -positive (FIG. 9A) GFAP-positive (FIG. 9B) and iNOS-positive (FIG. 9C) cells were counted in one section (two images per section) of each of six different mice (n=6) per group. Results represent analysis of one section of each of six different mice per group. *** p < 0.001. While at a dose of 8 mg / kg body wt / d, gemfibrozil markedly decreased the activation of microglia (FIG. 9A) and astroglia (FIG. 9B) and reduced the level of iNOS (FIG. 9C) in the SBF cortex of Cln3AJNCL mice, we did not find any inhibition of glial activation and drop in iNOS expression by gemfibrozil in Cln3AJNCLAPPARamice, suggesting that oral gemfibrozil is also unable to suppress glial inflammation in the absence of PPARa.

[0164] EXAMPLE 7:

[0165] Gemfibrozil improves locomotor activities in Cln3AJNCL mice via PPARa.

[0166] Since gemfibrozil treatment recovered locomotor activities in Cln3AJNCL mice as shown in (FIGs. 5A to 5F), we investigated whether such protection was also dependent on PPARa. Therefore, Cln3AJNCLAPPARamice were treated with gemfibrozil orally for 3 months followed by monitoring open field activities, pole test and rotorod performance. Turning to FIGs. 10A to 10G show the effect of gemfibrozil on locomotor activities of Cln3AJNCLAPPARa mice. Three-month old Cln3AJNCLAPPARa mice (n=6 per group) were treated with gemfibrozil (8 mg / kg body wt) daily via gavage. After 3 months of treatment, locomotor activities were monitored by open-field behavior (FIG. 10A, velocity; FIG. 10B, cumulative duration; FIG. 10C, distance travelled; FIG. 10D, center point moving), rotorod performance (FIG. 10E) and pole test (FIG. 10F, pole T-tum; FIG. 10G, pole latency). Results are mean + SEM of six mice per group. *** p < 0.001; NS, not significant. Consistent to the failure of gemfibrozil to reduce the accumulation of SCMAS and inhibit glial activation, gemfibrozil, in this instance as well, remained unable to improve overall open-field performance (not shown), velocity (FIG. 10 A), cumulative duration (FIG. 10B). distance travelled (FIG. 10C), and center point moving (FIG. 10D) of Cln3AJNCLAPPARtmice. Accordingly, gemfibrozil also did not improve the performance Cln3AJNCLAPPARamice on a moving rotorod (Fig. 10E) and pole descending (FIG. 10F and FIG. 10G), indicating that gemfibrozil enhances locomotor activities of Cln3AJNCL mice via PPARa.

[0167] Discussion

[0168] The neuronal ceroid lipofuscinoses (NCLs) or Batten disease are a group of recessively inherited fatal lysosomal storage disorders of infant, with rare adult forms (Kim et al., 2017; Johnson et al., 2019). Each form of NCL is caused by mutations in a different gene, which determines the age of disease onset, symptom and rate of disease progression, but all are fatal after a period of prolonged disability'. Other than Brineura therapy for LINCL, there are no effective therapies for NCL. Recently, different therapeutic approaches (e.g. enzyme replacement, immunosuppression, cell therapy, and gene therapy) are being considered for delaying or halting the progression of these devastating rare diseases. However, the transmembrane protein CLN3 is not secreted like either CLN1 or CLN2 and therefore, there is no option of its uptake by neighboring cells. Furthermore, since the disease process of JNCL is widespread in different parts of the brain, upon intracranial delivery, recombinant functional proteins or viral vectors may not reach the appropriate target cell type overcoming the densely populated. Although gene therapy trials are ongoing, development of neuroprotectivetherapeutic approaches for delaying the disease progression, improving locomotor functions and increasing the survival of JNCL patients are of paramount importance. Cln3Aex7 / 8 (Cln3 AJNCL) mice are useful in determining new therapeutic strategies and testing the efficacy of new drugs for JNCL. Here, we demonstrate for the first time that oral administration of gemfibrozil, an FDA-approved drug for hyperlipidemia in humans, reduces the CNS accumulation of storage material, decreases glial activation and improves locomotor functions in Cln3 AJNCL mice. Due to the increase in proinflammatory molecules in the CNS of ACln3 mice, there is the possibility that the motor impairments observed are due to sickness behavior, not structural alterations in the CNS. Although gemfibrozil treatment lowers the load of storage materials and improves the status of neurons as monitored by an increase in NeuN, further studies are needed to confirm that gemfibrozil-mediated improvement in locomotor performance in Cln3AJNCL mice is not due to suppression of neuroinflammation. Nevertheless, these results suggest that oral gemfibrozil may be beneficial for JNCL patients.

[0169] Accumulation of lipophilic and ceroid-like autofluorescent storage material in both neurons and nonneuronal cells is a signature feature of NCLs including JNCL. Mechanisms leading to the clearance of storage materials are becoming clear. Autophagy is the cell's way of removing unnecessary or dysfunctional components through a lysosomedependent degradation process to maintain cell homeostasis. Since aberrant lysosomal function and autophagy have been associated with multiple lysosomal storage and neurodegenerative disorders, upregulation of this lysosome - autophagy pathway has emerged as an attractive therapeutic strategy. TFEB is known as a master regulator of lysosomal biogenesis and several studies have shown that TFEB overexpression is capable alleviating neurodegenerative pathology' through upregulation of the autophagy-lysosome pathway. Recently we have taken a different approach to upregulate TFEB. We have demonstrated that Tfeb gene promoter harbors a consensus peroxisome proliferator response element (PPRE) and that activation of PPARa, neither PPAR|3 nor PPARy, leads to transcriptional upregulation of Tfeb. Since gemfibrozil is a known agonist of PPARa, we examined the role of this molecule and found that gemfibrozil treatment reduced CNS deposition of autofluorescent storage materials and exhibited neuroprotective effects via PPARa. Our conclusion is based on the followings: First, PPARa level was low in the SBF cortex of 6-month old Cln3AJNCL mice as compared to age- matched WT mice, which was increased and / or normalized by oral gemfibrozil. Second, JNCL disease process decreased the recruitment of PPARa to the Tfeb gene promoter as we found decreased enrollment of PPARa to Tfeb promoter in the SBF cortex of Cln3AJNCL mice in comparison to WT mice. However, gemfibrozil treatment was capable of stimulating PPARarecruitment to the Tfeb promoter in vivo in the CNS of Cln3AJNCL mice. Third, level of TFEB also decreased in the CNS of Cln3AJNCL mice that was increased by gemfibrozil treatment. Fourth, gemfibrozil was unable to decrease storage materials and improve locomotor performance in Cln3AJNCL mice lacking PPARa.

[0170] Similar to that found in other neurodegenerative disorders, activation of astrocytes and microglia is an early pathological event in human JNCL as well as Cln3 AJNCL mouse model of JNCL. Early-onset microgliosis has also been reported to closely accompany light induced retinal degeneration in Cln3AJNCL mice and the progressive loss of photoreceptor cells in the nclf mouse model of CLN6 disease. Here, we also found upregulation of GFAP and Iba-1 and associated expression of IL-ip, TNFa and iNOS in SBF cortex of Cln3 AJNCL mice. However, consistent to inhibition of proinflammatory molecules in cultured astrocytes and microglia, oral gemfibrozil treatment suppressed glial activation and inflammation in Cln3 AJNCL mice. However, gemfibrozil did not inhibit glial inflammation in Cln3AJNCL mice lacking PPARa, indicating an essential role of PPARa in this process. Suppressor of cytokine signaling (SOCS) proteins also play a crucial role in inhibiting cytokine signaling and inflammatory gene expression in various cell types, including glial cells. Similarly, IL-1R antagonist (IL-IRa) inhibits proinflammatory cell signaling by adhering to IL-1R, receptor for IL-la and IL-1 (3. We have seen that gemfibrozil is capable of upregulating both SOCS3 and IL-IRa in brain cells. Gemfibrozil treatment is also capable of increasing the level of SOCS3 and IL-IRa in striatum and cortex and exhibiting neuroprotection in Cln2(- / -) mice (Ghosh et al., 2017). Recently, we have demonstrated the involvement of PPARa in aspirin-induced transcription of both SOCS3 and IL-IRa in astrocytesi, indicating that gemfibrozil being a prototype agonist of PPARa, may also involve PPARa for the upregulation of SOCS3 and IL-IRa and hence exhibition of anti-inflammation.

[0171] Gemfibrozil has several advantages over other prospective neuroprotective agents. For example, gemfibrozil is an oral drug and fairly non-toxic. After oral administration, it can cross the blood-brain barrier. It is neuroprotective and can increase the lifespan of Cln2(_ / _)mice, an animal model of LINCL. Gemfibrozil has been reported to be safe for lowering lipids in children. Although primary site of CLN3 disease manifestation is the CNS, buildup of lysosomal storage material occurs in different part of the body and as a result, cardiac abnormality is also observed in children as the disease progresses. In this context, gemfibrozil may be able to prevent the cardiac disease in JNCL patients. Moreover, gemfibrozil exhibits neurotrophic and promyelinating effects and supports memory and learning.

[0172] In summary, this study demonstrates that oral administration of gemfibrozil, an FDA-approved lipid-lowering drug in humans, exhibits neuroprotective effects in an animal model of JNCL. In particular, gemfibrozil reduces storage materials, attenuates glial inflammation and improves locomotor activities in a mouse model of JNCL via PPARa, suggesting that gemfibrozil may not be neuroprotective for JNCL in the absence of PPARa. Although the in vivo state of Cln3AJNCL mice does not truly bear a resemblance to the in vivo scenario of JNCL patients and not much is known about the status of PPARa in JNCL, our results suggest that activation of PPARa by oral gemfibrozil may have therapeutic importance in JNCL.

[0173] All of the compositions and methods disclosed and claimed herein can be made and executed without undue experimentation in light of the present disclosure. While this invention may be embodied in many different forms, there are described in detail herein specific preferred embodiments of the invention. The present disclosure is an exemplification of the principles of the invention and is not intended to limit the invention to the particular embodiments illustrated.

[0174] Furthermore, the invention encompasses any and all possible combinations of some or all of the various embodiments described herein. It should also be understood that various changes and modifications to the presently preferred embodiments described herein will be apparent to those skilled in the art. Such changes and modifications can be made without departing from the spirit and scope of the invention and without diminishing its intended advantages. It is therefore intended that such changes and modifications be covered by the appended claims.

Claims

CLAIMS:

1. A method for treating a patient diagnosed with or suspected of having juvenile neuronal ceroid lipofuscinosis (JNCL) in need thereof, comprising administering to the patient a composition comprising a therapeutically effective amount of gemfibrozil, or a pharmaceutically acceptable salt thereof, wherein the administering results in a reduction in symptoms associated with JNCL, thereby treating the patient.

2. The method of claim 1, wherein the patient is at least about 6 months old.

3. The method of claim 1, wherein the patient is at least about 3 years old.

4. The method of claim 1. wherein the patient is between about 6 months to about10 years old.

5. The method of claim 1, wherein the composition further comprises at least one pharmaceutically acceptable excipient or carrier.

6. The method of claim 1. wherein the composition is formulated to be administered orally to the patient.

7. The method of claim 1 , wherein the patient is not displaying symptoms of JNCL before administering the composition.

8. The method of claim 1, wherein the composition is administered at least once daily.

9. The method of claim 1, wherein the composition is administered at least twice daily.

10. The method of claim 1 further comprising co-administering a second therapy.

11. The method of claim 10, wherein the second therapy is a gene therapy.

12. A composition comprising gemfibrozil, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for treating JNCL in a patient.

13. The composition of claim 12, wherein the composition is formulated to be given orally to the patient.

14. The composition of claim 12, wherein the patient is between the ages of about 3 to about 10 years old.

15. The composition of claim 12, wherein the patient is at least about 6 months old.

16. A composition comprising gemfibrozil, or a pharmaceutically acceptable salt thereof, for us in a therapy to treat JNCL in a patient.

17. The composition of claim 16, wherein the composition is formulated to be given orally to the patient.

18. The composition of claim 17, wherein the patient is between the ages of about 3 to about 10 years old.

19. The composition of claim 18, wherein the patient is at least about 6 months old.

20. A method for treating or correcting a defective lysosomal transmembrane glycoprotein Cln3 in a patient in need thereof, comprising administering to a patient in need thereof an effective amount of a pharmaceutical composition comprising gemfibrozil and at least one pharmaceutically acceptable excipient or carrier.

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