TPP1 formulations and methods for treating cln2 disease

Abstract

Disclosed are formulations comprising recombinant human tripeptidyl peptidase-1 (rhTPP1) for intrathecal, intracerebroventricular, or intraocular administration, and kits comprising the formulations. Also disclosed are methods of using rhTPP1 to prevent and treat symptoms of neuronal ceroid lipofuscinosis (CLN2) disease. The formulations and methods are effective to halt progression of CLN2 disease and can be used to treat individuals with CLN2 or a family history of CLN2.

Description

[0001] Cross-references to related applications

[0002] Pursuant to 35 U.S. SC §119(e), the benefits of U.S. Provisional Patent Application No. 62 / 300,171, filed February 26, 2016, and U.S. Provisional Patent Application No. 62 / 158,789, filed May 8, 2015, are claimed herein, the disclosures of which are incorporated herein by reference in their entirety.

[0003] sequence list

[0004] This application contains a sequence list in computer-readable form (49443_SeqListing.txt; 8,137 bits; created April 11, 2016), which is incorporated herein by reference in its entirety as another part of this invention. Technical Field

[0005] This invention relates to formulations comprising recombinant human tripeptidyl peptidase-1 and their use in treating neuronal ceroid lipofuscinosis and related physiological symptoms. Background Technology

[0006] Neuronal cerebrolipofuscinosis (CLN2) is a rare genetic disorder characterized by a deficiency of the lysosomal enzyme tripeptidyl peptidase-1 (TPP1) caused by a mutation in the TPP1 gene. CLN2 is an autosomal recessive disorder with an estimated incidence of 0.5 per 100,000 births. In the absence of TPP1, lysosomal storage substances normally metabolized by this enzyme accumulate in many organs, and in the central nervous system, this accumulation causes the neurodegenerative symptoms typical of CLN2. The progression of untreated CLN2 is well characterized, and the natural history of the disease is highly consistent and predictable, as evidenced by natural history data from independent patient populations in North America and Europe.

[0007] CLN2 disease exhibits a distinctly 'classic' late-onset infantile phenotype. Children with CLN2 typically develop normally until around age 3, when the first symptoms appear. Most commonly, CLN2 patients will experience their first unexplained seizures, and the acquisition of normal language milestones will begin to lag. By age 3, all patients will exhibit one or more of the signs of the disease, including, for example, seizures, dementia, loss of movement, motor impairment, blindness, clumsiness, ataxia, and cognitive decline. From the onset of clinical symptoms, the disease progresses rapidly and aggressively, generally causing complete loss of language, cognition, gait, fine motor skills, medullary function, and vision within 2 to 4 years, leading to immobility, aphonia, and blindness. Patients remain in a vegetative state until death, which typically occurs between the ages of 6 and 12.

[0008] Two quantitative rating scales have been developed by specialist clinicians to assess the severity of CLN2 disease and have been used in natural history studies: (1) the Hamburg scale (Steinfeld et al., *American Journal of Medical Genetics*, 2002; 112(4):347-54); and (2) the Weill Cornell Medical College (WCMC) scale (Worgall et al., *Neurology*, 2007; 69(6):521-35). The two scales have similar structures and assessment methods. Both scales measure the loss of previously acquired important neurological milestones in CLN2 patients, with the lost units on the disease rating scale representing fundamental milestones in progressive decline.

[0009] An analysis of the disease course in untreated children with CLN2 showed a predictable loss of all language and gait within 3 years of disease onset, with an average loss of 2.1 milestone events per year (i.e., a loss of 2.1 points on the disease rating scale). Language decline typically precedes gait, so the first year is characterized by intelligible speech loss progressing to ataxic gait, the second year by loss of walking and intelligible language, and the third year by loss of any motor or communication function.

[0010] Recombinant human tripeptidyl peptidase-1 (rhTPP1) is being developed as a potential treatment for CLN2 disease. The rhTPP1 protein is produced in cell cultures as a proenzyme, lacking enzymatic activity. The proenzyme is autoactivated upon uptake into lysosomes at acidic pH (and via lysosomal proteases). The mature native TPP1 protein is a lysosomal serine protease and the only known mammalian member of the sedolisin (serine-carboxypeptidase) family, characterized by a highly conserved Ser-Glu-Asp (SED) catalytic triad. The catalytic triad on rhTPP1 is formed by S456, E253, and D341. The enzyme's primary activity is as a tripeptidyl peptidase with broad acceptor specificity. The enzyme's activity on its acceptors results in the continuous release of tripeptides from the N-terminus of the protein acceptor (Oyama et al., *Journal of Biochemistry*, 2005; 138(2):127-34). Secondary, significantly weaker internal protein cleavage activity has also been reported at a pH of 3 (Lin et al., Journal of Biol Chem., 2001; 276(3):2249-55).

[0011] The only commercially available treatment for CLN2 is symptomatic and supportive care; currently, there are no approved treatment options to slow or halt the relentless progression of CLN2, let alone reverse the detrimental effects of the disease (Mole, SE and Williams, RE, 2010, Gene Reviews; Chang et al., The Neuronal Ceroid Lipofuscinoses (Batten Disease); 2011, Oxford University Press). Preserving these children's motor, language, and / or visual abilities has clinically significant benefits for both patients and parents / caregivers. Therefore, there is a need for new treatments for CLN2 that reduce or prevent the physiological decline associated with the disease. Summary of the Invention

[0012] This invention relates to formulations, kits, methods, and medical uses of recombinant human tripeptidyl peptidase-1 (rhTPP1), specifically for the treatment of neuronal ceroid lipofuscinosis (CLN2) and the treatment or prevention of one or more physiological symptoms associated with the onset of said disease. In one aspect, the invention provides a formulation comprising rhTPP1 for intraventricular, intrathecal, or intraocular administration. Optionally, rhTPP1 comprises the amino acid sequence of SEQ ID NO:1 or SEQ ID NO:2, or a fragment thereof, for example, at a concentration of about 30 mg / mL in the formulation. The formulation may have a pH of about 6.5. The formulation may further include any one of the following: potassium chloride at a concentration of about 0.01 mg / mL to about 1 mg / mL; magnesium chloride hexahydrate at a concentration of about 0.01 mg / mL to about 1 mg / mL; calcium chloride dihydrate at a concentration of about 0.01 mg / mL to about 1 mg / mL; disodium hydrogen phosphate heptahydrate at a concentration of about 0.01 mg / mL to about 1 mg / mL; sodium dihydrogen phosphate monohydrate at a concentration of about 0.01 mg / mL to about 1 mg / mL; sodium chloride at a concentration of about 1 mg / mL to about 20 mg / mL; or any one or a combination of the above. Optionally, the formulation is free of preservatives and / or stable at about 5°C for, for example, at least about 6 months. In one embodiment, the formulation comprises rhTPP1 at a concentration of about 30 mg / mL, disodium hydrogen phosphate heptahydrate at a concentration of about 0.11 mg / mL, sodium dihydrogen phosphate at a concentration of about 0.08 mg / mL, sodium chloride at a concentration of about 8.77 mg / mL, potassium chloride at a concentration of about 0.22 mg / mL, magnesium chloride at a concentration of about 0.16 mg / mL, and calcium chloride dihydrate at a concentration of about 0.21 mg / mL.

[0013] In another aspect, the present invention provides a method for treating CLN2 disease or one or more symptoms associated with CLN2 disease, comprising administering to an individual in need about 10 mL of a composition comprising a dose of about 300 mg of rhTPP1, wherein the composition is administered to the individual over a period of about 4 hours. The present invention also provides a composition comprising rhTPP1 for treating an individual with CLN2 disease or one or more symptoms associated with CLN2 disease, comprising about 10 mL of a composition comprising a dose of about 300 mg of rhTPP1, wherein the dose is administered to the individual over a period of about 4 hours. The present invention also provides the use of rhTPP1 in the manufacture of an agent for treating CLN2 disease or one or more symptoms associated with CLN2 disease, wherein the agent comprises about 10 mL of a composition comprising a dose of about 300 mg of rhTPP1, wherein the dose is administered to the individual over a period of about 4 hours. In one aspect, the composition comprising rhTPP1 is administered to the individual via a catheter. Optionally, the catheter is flushed after administration of the composition by administering up to about 5 mL, preferably about 3 mL, more preferably about 2 mL, of a flushing solution. The present invention also provides a method for preventing or delaying the onset of one or more symptoms of CLN2 disease, comprising administering to an individual in need about 10 mL of a composition comprising a dose of about 300 mg of rhTPP1, optionally wherein the individual has a family history of CNL2 disease and wherein the composition is administered to the individual over a period of about 4 hours. In one aspect, the composition comprising rhTPP1 is administered to the individual via a catheter. After administration of the composition, the catheter is optionally flushed with a flushing solution administered in an amount of up to about 5 mL, preferably about 3 mL, more preferably about 2 mL. The composition may be administered intrathecally, intraventricularly, and / or intraocularly, and optionally every week. Optionally, the composition is administered intraventricularly or intrathecally, and is administered subcephalicly without removing cerebrospinal fluid from the individual just before administration of the composition comprising rhTPP1. In another embodiment, the composition may be administered in an isovolumetric manner, i.e., a defined volume of CSF is removed from the patient, the volume of which is approximately equal to the volume of the composition subsequently administered intrathecally.

[0014] In one aspect, the present invention provides a method for treating CLN2 disease or one or more symptoms associated with said disease, comprising administering rhTPP1 to an individual in need at a dose and / or frequency that effectively maintains or slows or reduces the decline in physiological function, wherein said physiological function is language function, motor function, vision, or eating function. The present invention also provides a composition comprising rhTPP1 for treating CLN2 disease or one or more symptoms associated with said disease, comprising a dose of rhTPP1 that effectively maintains or slows or reduces the decline in physiological function, wherein said physiological function is language function, motor function, vision, or eating function. The present invention also provides the use of rhTPP1 in the manufacture of an agent for treating CLN2 disease or one or more symptoms associated with said disease, wherein said agent comprises a dose of rhTPP1 that effectively maintains or slows or reduces the decline in physiological function, wherein said physiological function is language function, motor function, vision, or eating function.

[0015] In another aspect, the present invention provides a method for treating an individual with CLN2 disease or one or more symptoms associated with said disease, comprising administering rhTPP1 to the individual in need at a dose and / or frequency that effectively improves physiological function in the individual, wherein said physiological function is language function, motor function, vision, or eating function. The present invention also provides a composition comprising rhTPP1 for treating an individual with CLN2 disease or one or more symptoms associated with said disease, comprising a dose of rhTPP1 that effectively improves physiological function in the individual, wherein said physiological function is language function, motor function, vision, or eating function. The present invention also provides the use of rhTPP1 in the manufacture of an agent for treating CLN2 disease or one or more symptoms associated with said disease, wherein said agent comprises a dose of rhTPP1 that effectively improves physiological function in the individual, wherein said physiological function is language function, motor function, vision, or eating function. In another aspect, the present invention provides a method for treating an individual with CLN2 disease or one or more symptoms associated with said disease, comprising administering rhTPP1 to the affected individual at a dose and / or frequency effective in preventing or treating neurological symptoms of said disease, wherein said neurological symptoms are decreased brain volume, reduced brain gray matter, seizures, or increased cerebrospinal fluid. The present invention also provides a composition comprising rhTPP1 for treating an individual with CLN2 disease or one or more symptoms associated with said disease, comprising a dose of rhTPP1 effective in preventing or treating neurological symptoms of said disease, wherein said neurological symptoms are decreased brain volume, reduced brain gray matter, seizures, or increased cerebrospinal fluid. The present invention also provides the use of rhTPP1 in the manufacture of an agent for treating CLN2 disease or one or more symptoms associated with said disease, wherein said agent comprises a dose of rhTPP1 effective in preventing or treating neurological symptoms of said disease, wherein said neurological symptoms are decreased brain volume, reduced brain gray matter, seizures, or increased cerebrospinal fluid. Optionally, the individual has a family history of CLN2 disease.

[0016] In another aspect, the present invention provides a method for preventing or reducing CLN2-related motor / gait degeneration in an individual, comprising administering a therapeutically effective dose of rhTPP1 to said individual. The present invention also provides a composition comprising a therapeutically effective dose of rhTPP1 for preventing or reducing CLN2-related motor / gait degeneration in an individual. The present invention also provides the use of a therapeutically effective dose of rhTPP1 for manufacturing a medicament for preventing or reducing CLN2-related motor / gait degeneration in an individual. In one aspect, the method comprises administering a dose of rhTPP1 that effectively prevents or reduces a decline in an individual's clinical disease rating, as measured using the WCMC Gait Disease Rating Scale or the Hamburger Disease Rating Scale for Motor Skills, compared to a previous rating determined before or during treatment.

[0017] In another aspect, the present invention provides a method for preventing or reducing CLN2-related language regression in an individual, comprising administering a therapeutically effective dose of rhTPP1 to said individual. The present invention also provides a composition comprising a therapeutically effective dose of rhTPP1 for preventing or reducing CLN2-related language regression in an individual. The present invention also provides the use of a therapeutically effective dose of rhTPP1 for manufacturing a medicament for preventing or reducing CLN2-related language regression in an individual. In one aspect, the method comprises administering a dose of rhTPP1 that effectively prevents or reduces regression in an individual's clinical disease rating, as measured, for example, by the WCMC or Hamburger Disease Rating Scale for Language Use, compared to a previous rating determined before or during treatment.

[0018] In another aspect, the present invention provides a method for preventing or reducing CLN2-related visual impairment in an individual, comprising administering a therapeutically effective dose of rhTPP1 to said individual. The present invention also provides a composition comprising a therapeutically effective dose of rhTPP1 for preventing or reducing CLN2-related visual impairment in an individual. The present invention also provides the use of a therapeutically effective dose of rhTPP1 for manufacturing a medicament for preventing or reducing CLN2-related visual impairment in an individual. In one aspect, the method comprises administering a dose of rhTPP1 that effectively prevents or reduces a decline in an individual's clinical disease rating, as measured, for example, using the Hamburger Disease Rating Scale for Visual Function, compared to a previous rating determined before or during treatment.

[0019] In another aspect, the present invention provides a method for preventing or reducing CLN2-related brain volume degeneration in an individual, comprising administering a therapeutically effective dose of rhTPP1 to the individual. The present invention also provides a composition comprising a therapeutically effective dose of rhTPP1 for preventing or reducing CLN2-related brain volume degeneration in an individual. The present invention also provides the use of a therapeutically effective dose of rhTPP1 for manufacturing a medicament for preventing or reducing CLN2-related brain volume degeneration in an individual. In one aspect, the method comprises administering a dose of rhTPP1 that effectively prevents or reduces a decrease in brain volume and / or gray matter volume compared to a previous volume determined before or during treatment.

[0020] In one aspect, the present invention provides a kit comprising a formulation containing rhTPP1 as described herein and a flushing solution. The flushing solution may constitute any formulation of the present invention, wherein rhTPP1 is omitted. For example, the flushing solution may comprise about 0.11 mg / mL of disodium hydrogen phosphate heptahydrate, about 0.08 mg / mL of sodium dihydrogen phosphate monohydrate, about 8.77 mg / mL of sodium chloride, about 0.22 mg / mL of potassium chloride, about 0.16 mg / mL of magnesium chloride hexahydrate, and about 0.21 mg / mL of calcium chloride dihydrate. Optionally, the kit further comprises a reservoir and catheter for implantation. Optionally, the kit may comprise one or more elements selected from the group consisting of: extension tubing, in-line filter, port needle, at least one (optionally two) syringe, at least one (optionally two) syringe needle, and combinations thereof.

[0021] The above summary is not intended to define every aspect of the invention, and other features and advantages of the invention will become apparent from the following detailed description, including the drawings. This invention is intended to be a unified document, and it should be understood that it covers all combinations of features described herein, even if such combinations are not found together in the same sentence, paragraph, or section of the invention. Furthermore, the invention includes all embodiments of the invention whose scope is, in any case, narrower than the variations specifically mentioned above, as another aspect. Unless the context clearly requires a more limiting meaning, the term "a / an" in the description or claim of aspects of the invention should be understood to mean "one or more". Regarding the description of one or more elements within a device, it should be understood that it covers all combinations within the device. If an aspect of the invention is referred to as "comprising" a feature, then it also covers embodiments "consisting of" or "substantially consisting of" the features. Other features and variations of the invention will be apparent to those skilled in the art from the entirety of this application, and all such features are intended to be aspects of the invention. Attached Figure Description

[0022] Figure 1 The amino acid sequence of the rhTPP1 proenzyme lacking the associated signal peptide is depicted. The pre-enzyme segment consists of the first 176 amino acid residues, and the mature enzyme is 368 amino acids long starting from position 177.

[0023] Figure 2 This study depicts the clinical progression of untreated individuals with CLN2 disease in a natural history study and shows the Hamburger motor and language integration scores from 0 to 6 as a function of patient age. Median, interquartile, and 10% / 90% distributions are presented, except for the mean and 95% confidence intervals.

[0024] Figures 3A to 3FClinical assessments of 24 patients generated during the treatment duration are depicted, and Hamburger Motor and Verbal Syndrome (CLN2) scores ranging from 0 to 6 are presented. Hollow circles represent CLN2 scores obtained at or before the first 300 mg rhTPP1 infusion, while closed circles represent CLN2 scores obtained after the first 300 mg rhTPP1 infusion. The influence of the total score (circles) on the motor / gait (squares) and verbal (triangles) scores is shown. Day 1 of the analysis is the date of the first infusion.

[0025] Figures 4A to 4I The changes in CLN2 scores were compared between nine patients treated with rhTPP1 and matched untreated natural history patients, using disease rating scores (denoted by the prefix "HAM") based on the Hamburger Motor and Language Subscale (0-6). In each figure, solid lines represent the outcomes of treated patients, compared with dashed lines representing the outcomes of matched untreated natural history patients.

[0026] Figure 5 The distribution of clinical changes from baseline is depicted in matched untreated natural history patients (circles) during the duration of treatment matched with the study individuals (squares).

[0027] Figures 6A to 6I The changes in CLN2 scores from nine patients treated with rhTPP1 to untreated matched natural history patients are depicted based on disease rating scores using the Hamburg Motor / Verbal / Visual Total Subscale (0-9). In each figure, solid lines represent the outcomes of the treated patients, compared to dashed lines representing the outcomes of the untreated matched natural history patients.

[0028] Figure 7 The volume (top plot) and proportion (bottom plot) of cerebrospinal fluid measured during the treatment duration are depicted for all 24 patients. Each line represents one patient.

[0029] Figure 8A to 8L The brain volumes of 24 treated patients are depicted. The volume (top plot) and proportion (bottom plot) of white matter are shown as the difference between the total brain volume (dotted line) and the CSF and gray matter (dashed line), and the volume and proportion of gray matter are shown as the difference between the CSF and gray matter (dashed line) and CSF (solid line).

[0030] Figure 9A and 9B The mean changes in CLN2 scores were depicted in patients treated with rhTPP1 and in untreated patients with a natural history. Figure 9A The CLN2 scores of 23 patients (dashed line) treated with 300 mg rhTPP1 for 48 weeks and 41 individuals in the untreated natural history group (solid line) are depicted. Figure 9BThe changes in CLN2 scores from baseline were depicted in 23 patients treated with 300 mg rhTPP1 for 48 weeks.

[0031] Figures 10A to 10L Clinical assessments of 24 patients generated during the treatment duration are depicted, showing the sum of scores for hamburger (left panel) motor function (square), speech (triangle), tics (cross), and visual acuity (diamond) from 0 to 12, and the sum of scores for WCMC (right panel) gait (square), speech (triangle), myoclonus (cross), and eating (diamond) from 0 to 12. Hollow circles represent the total CLN2 score obtained at or before the first 300 mg rhTPP1 infusion, while closed circles represent the total CLN2 score obtained after the first 300 mg rhTPP1 infusion. Detailed Implementation

[0032] The following definitions are intended to assist skilled practitioners in understanding this invention. Unless otherwise defined herein, the scientific and technical terms used in this invention should have meanings commonly understood by one of ordinary skill in the art. Within the range of values ​​provided, it should be understood that each interpolated value between the upper and lower limits of the range, and between any other stated or interpolated values ​​within the range, is encompassed within this invention, accurate to one-tenth of the lower limit unit unless the context clearly specifies otherwise. The upper and lower limits of these smaller ranges may be independently included within the smaller range, subject to any specific exclusions within the range.

[0033] The term "family history" refers to an individual whose blood relatives, such as brothers, parents, grandparents, great-grandparents, or great-great-grandmothers, have been diagnosed with CLN2 disease.

[0034] In one aspect, the term "fragment" refers to a sequence including SEQ ID NO:1 and Figure 1The fragment is a recombinant protein comprising a portion of the amino acid sequence of the rhTPP1 preenzyme described in SEQ ID NO:1. For example, the fragment may contain at least about 60%, at least about 70%, at least about 80%, at least about 90%, or at least about 95% of the amino acid sequence described in SEQ ID NO:1. In another aspect, the fragment may comprise the full-length (368 amino acids long; amino acids 177-544 of SEQ ID NO:1) mature TPP1 enzyme amino acid sequence described in SEQ ID NO:2, a portion thereof, and / or a catalytic triplet formed by amino acid residues S456, E253, and D341. The fragment retains catalytic activity. For example, the fragment exhibits tripeptidyl peptidase activity and / or exhibits catalytic activity that causes the continuous release of tripeptides from the N-terminus of the protein acceptor. In some aspects, the "fragment" of the rhTPP1 proenzyme comprises at least 500 consecutive amino acids of SEQ ID NO:1, at least 450 consecutive amino acids of SEQ ID NO:1, at least 400 consecutive amino acids of SEQ ID NO:1, at least 368 amino acids of SEQ ID NO:1, at least 350 amino acids of SEQ ID NO:1, or at least 300 consecutive amino acids of SEQ ID NO:1. In other aspects, the "fragment" of the rhTPP1 proenzyme comprises at least 350 consecutive amino acids of SEQ ID NO:2, at least 325 consecutive amino acids of SEQ ID NO:2, at least 300 consecutive amino acids of SEQ ID NO:2, at least 275 amino acids of SEQ ID NO:2, at least 250 amino acids of SEQ ID NO:2, or at least 200 consecutive amino acids of SEQ ID NO:2.

[0035] The term "ventricle" refers to a ventricular system (e.g., a ventricle) in which the composition is administered, for example, by injection, infusion, or implantation.

[0036] The term "intraocular" refers to the administration of a composition, for example by injection, infusion, or implantation into an ocular region (e.g., within the eyeball) or topically / to the eye (e.g., using a cream, ointment, gel, or drop).

[0037] The term "intrathecal" refers to the administration of the composition, for example, via injection, infusion, or implantation, to the lumbar region (e.g., the subarachnoid space of the spinal cord).

[0038] The term "therapeutic efficacy" refers to any therapeutic benefit resulting from the treatment methods of the present invention. For example, such an effect can be a beneficial effect manifested in appropriate target tissues or organs, wherein such beneficial physiological effects are compared to physiological parameters measured in the absence of enzyme replacement therapy. Such a therapeutic effect can be any reduction or elimination of one or more clinical or subclinical manifestations of CLN2 disease. For example, therapeutic efficacy improves, reverses, delays, prevents, or reduces one or more physiological functional declines and / or neurological symptoms of CLN2 as described herein.

[0039] The term "stable" or "stabilized" refers to a protein-containing formulation in which the protein components substantially retain their physical, functional, and / or chemical stability over time during storage. Stability can be measured at a selected temperature for a selected time period. Preferably, the formulation is stable for at least one month at room temperature (about 30°C) or at about 40°C, and / or stable for at least one year, and preferably at least two years, at about 2°C to about 8°C. For example, the degree of protein degradation or aggregation during storage can be used as an indicator of protein stability. Thus, a "stable" formulation can be one in which less than about 20%, more preferably less than about 10%, and most preferably less than about 5% of the protein components are present in the formulation in a degraded or aggregated form after storage. A "stable" formulation substantially retains the same functional or therapeutic characteristics as a freshly prepared formulation. Various analytical techniques for measuring protein stability are applicable in the field and are reviewed, for example, in Peptide and Protein Drug Delivery, 247-301, edited by Vincent Lee, Marcel Dekker, Inc., New York, NY, Pubs. (1991) and Jones, A. Adv. Drug Delivery Rev. 10:29-90 (1993).

[0040] When referring to the prevention or reduction of one or more symptoms or physiological consequences of CLN2 disease in an individual, the terms "prevention" or "reduction" or their grammatical equivalents mean that the rate of decline of the one or more symptoms in a treated CLN2 individual is slower than that observed in an untreated CLN2 individual. In this regard, untreated CLN2 can be the same individual subsequently treated with the compositions of the present invention, or it can be the average rate of decline of the relevant symptoms as observed from the results of natural history studies disclosed herein.

[0041] Under jurisdictions where methods performed on humans are prohibited from being patented, "administering" rhTPP1 or a formulation thereof to a human individual means medical use of rhTPP1 or a formulation thereof, such as the use of rhTPP1 or a formulation thereof to treat the CLN2 disease described herein, or the use of rhTPP1 to manufacture an agent for treating the CLN2 disease as described herein. The broadest reasonable interpretation, consistent with the law or regulation defining the patentable subject matter, is intended. Under jurisdictions where methods performed on humans are prohibited from being patented, "administering" rhTPP1 or a formulation thereof includes methods performed on humans and the aforementioned activities.

[0042] This invention provides formulations and kits comprising rhTPP1, and methods for treating CLN2 disease using them. Administration of rhTPP1 allows the protein to be absorbed by cells throughout the central nervous system via a cation-independent mannose-6-phosphate receptor (CI-MPR) and localized to lysosomes within cells. The enzyme's absorption into lysosomes and subsequent activation promotes increased catabolic metabolism of stored substances in diseased tissues, reduces the gradual accumulation of lysosomal stored substances, and prevents disease progression. The formulations and methods of this invention provide therapeutic benefits superior to currently approved treatments.

[0043] Mixtures

[0044] {0044} This invention provides formulations comprising rhTPP1 for intraventricular, intrathecal, or intraocular administration. In one aspect, rhTPP1 comprises SEQ ID NO:1 or a fragment thereof. The RhTPP1 protein applicable to the formulations and methods described herein, and methods for obtaining the rhTPP1 protein, are described in U.S. Patent Nos. 6,302,685 and 8,277,800, which are incorporated herein by reference in their entirety.

[0045] In one aspect, rhTPP1 includes the amino acid sequence of SEQ ID NO:1 ( Figure 1 The amino acid sequence shown contains amino acids 1-544) or fragments thereof that are catalytically active. In another aspect, rhTPP1 comprises the amino acid sequence of SEQ ID NO:2 ( Figure 1The amino acid sequence shown is amino acids 177-544) or fragments thereof that have catalytic activity. In another aspect, rhTPP1 has detectable enzymatic activity or is processed in vivo into an enzyme form with detectable enzymatic activity (i.e., "functional") and has at least about 70% sequence identity with SEQ ID NO:1 or SEQ ID NO:2. For example, functional rhTPP1 is at least about 70% identical, at least about 75% identical, at least about 80% identical, at least about 85% identical, at least about 90% identical, at least about 95% identical, or at least about 97% identical with SEQ ID NO:1 or SEQ ID NO:2. In one aspect, the formulation is a liquid formulation comprising rhTPP1 at concentrations from about 1 mg / mL to about 100 mg / mL, for example, from about 10 mg / mL to about 50 mg / mL, from about 25 mg / mL to about 40 mg / mL, or from about 30 mg / mL to about 60 mg / mL. In all aspects, the formulation includes rhTPP1 at concentrations of about 1 mg / mL to about 100 mg / mL, about 5 mg / mL to about 80 mg / mL, about 10 mg / mL to about 50 mg / mL, about 20 mg / mL to about 40 mg / mL, about 25 mg / mL to about 35 mg / mL, and more specifically, about 1 mg / mL, about 10 mg / mL, about 20 mg / mL, about 30 mg / mL, about 40 mg / mL, about 50 mg / mL, about 60 mg / mL, about 70 mg / mL, about 80 mg / mL, about 90 mg / mL, or about 100 mg / mL. In one aspect, the formulation has a pH of about 5.5 to about 7.5 or about 6.0 to about 7.0, for example, about 5.5, about 6.0, about 6.5, about 7.0, or about 7.5.

[0046] In one aspect, the formulations of the present invention comprising rhTPP1 further comprise one or more excipients for maintaining the levels of key electrolytes in cerebrospinal fluid (CSF) or ocular fluid. For example, in one aspect, in addition to rhTPP1 or fragments thereof, the formulation further comprises potassium chloride at concentrations of about 0.01 mg / mL to about 1 mg / mL, such as about 0.1 mg / mL to about 0.5 mg / mL, about 0.2 mg / mL to about 0.8 mg / mL, about 0.2 mg / mL to about 0.4 mg / mL, about 0.15 mg / mL to about 0.25 mg / mL, or about 0.05 mg / mL to about 0.3 mg / mL. In another aspect, the formulation further comprises magnesium chloride hexahydrate at a concentration of about 0.01 mg / mL to about 1 mg / mL, for example, about 0.1 mg / mL to about 0.5 mg / mL, about 0.1 mg / mL to about 0.8 mg / mL, about 0.1 mg / mL to about 0.3 mg / mL, about 0.15 mg / mL to about 0.25 mg / mL, or about 0.05 mg / mL to about 0.3 mg / mL. In another aspect, the formulation further comprises calcium chloride dihydrate at a concentration of about 0.01 mg / mL to about 1 mg / mL, for example, about 0.1 mg / mL to about 0.5 mg / mL, about 0.2 mg / mL to about 0.8 mg / mL, about 0.15 mg / mL to about 0.25 mg / mL, about 0.1 mg / mL to about 0.3 mg / mL, or about 0.05 mg / mL to about 0.3 mg / mL. In another aspect, the formulation includes a combination of all or any of the above.

[0047] In another aspect, formulations including rhTPP1 further include one or more buffers. For example, in various aspects, the formulation further includes disodium hydrogen phosphate heptahydrate at a concentration of about 0.01 mg / mL to about 1 mg / mL, such as about 0.1 mg / mL to about 0.5 mg / mL, about 0.05 mg / mL to about 0.4 mg / mL, or about 0.1 mg / mL to about 0.3 mg / mL; and / or sodium dihydrogen phosphate monohydrate at a concentration of about 0.01 mg / mL to about 1 mg / mL, such as about 0.01 mg / mL to about 0.2 mg / mL, about 0.05 mg / mL to about 0.3 mg / mL, or about 0.08 mg / mL to about 0.4 mg / mL.

[0048] In another aspect, the formulation further comprises an isotensive agent, such as sodium chloride, at concentrations of about 1 mg / mL to about 20 mg / mL, for example, about 1 mg / mL to about 10 mg / mL, about 5 mg / mL to about 15 mg / mL, or about 8 mg / mL to about 20 mg / mL. Other buffers and isotensive agents known in the art are suitable and can generally be used in the formulations of the present invention.

[0049] In one aspect, the formulation comprising about 30 mg / mL of rhTPP1 further comprises about 0.11 mg / mL of disodium hydrogen phosphate heptahydrate, about 0.08 mg / mL of sodium dihydrogen phosphate monohydrate, about 8.77 mg / mL of sodium chloride, about 0.22 mg / mL of potassium chloride, about 0.16 mg / mL of magnesium chloride hexahydrate, about 0.21 mg / mL of calcium chloride dihydrate, and a diluent, such as water for injection.

[0050] The rhTPP1 formulation of the present invention is stable and can be stored for extended periods without unacceptable changes in quality, efficacy, or purity. In one aspect, the formulation is stable at a temperature of about 5°C (e.g., 2°C to 8°C) for at least one month, for example, at least one month, at least three months, at least six months, at least 12 months, at least 18 months, at least 24 months, or more. In another aspect, the formulation is stable at a temperature less than or equal to about -20°C for at least six months, for example, at least six months, at least 12 months, at least 18 months, at least 24 months, at least 36 months, or more. In yet another aspect, the formulation is stable at a temperature less than or equal to about -40°C for at least six months, for example, at least six months, at least 12 months, at least 18 months, at least 24 months, at least 36 months, or more. In yet another aspect, the formulation is stable at a temperature less than or equal to about -60°C for at least six months, for example, at least six months, at least 12 months, at least 18 months, at least 24 months, at least 36 months, or more.

[0051] In one aspect, the formulations of the present invention are free of preservatives and / or stabilizers, and therefore do not contain thimerosal, phenylmercuric salts, chlorhexidine, phenol, benzoic acid, sorbic acid, parabens, alcohols, or other preservatives commonly found in non-intestinal or ocular formulations.

[0052] In another aspect, the formulations of the present invention may include one or more preservatives, stabilizers, or excipients. In this regard, many well-known and commonly used preservatives, stabilizers, and excipients suitable for intrathecal or ICV delivery of protein-containing formulations are known in the art. More specifically, examples of such additives for intrathecal or ICV delivery of enzyme-containing formulations are described in WO2013 / 096899, which is incorporated herein by reference.

[0053] method

[0054] This invention provides a method for treating CLN2 disease, the method comprising administering a therapeutically effective amount of the formulation comprising rhTPP1 described herein to an individual in need. The invention also provides a composition comprising rhTPP1 for treating the CLN2 disease described herein, and use of rhTPP1 in the manufacture of a medicament for treating the CLN2 disease described herein. In one aspect, the severity and progression of CLN2 disease and the therapeutic benefit of administering rhTPP1 to a patient can be measured using the Hamburger or WCMC Clinical Disease Rating Scale. The Hamburger and WCMC scales consist of four disease-related domains, which are rated on a scale of 0 to 3, where 3 is normal, 2 is abnormal but functional, 1 is abnormal and significantly impaired, and 0 has no functional remainder. Two of the four domains, gait / motor and language, are shared between the two scales and have high inherent content accuracy. Each scale generally captures changes present as a function of disease progression and disease management. Gait, language, and visual acuity scales capture disease progression. The frequency of seizures, motor disturbances, and feeding are determined by nursing care, particularly anticonvulsant medication and feeding tube management. Clinical progression is often assessed using the overall language and gait subscales, with a score of 6 representing age-normal and 0 representing complete loss of function. Table 1 describes the WCMC and Hamburg CLN2 disease scales.

[0055] Table 1

[0056]

[0057]

[0058] In various aspects, the present invention provides a method for treating CLN2 disease or one or more clinical symptoms of CLN2 disease, comprising administering to an individual in need a composition comprising a therapeutically effective amount of rhTPP1; use of rhTPP1 in the manufacture of a medicament for treating an individual's CLN2 disease, or use of rhTPP1 for treating an individual's CLN2 disease.

[0059] The present invention also provides a method for preventing one or more clinical symptoms of CLN2 disease, comprising administering to an individual in need a formulation comprising rhTPP1, optionally wherein said individual has a family history of CLN2 disease. In various aspects, the present invention provides a method for preventing one or more clinical symptoms of CLN2 disease, comprising administering to an individual in need a composition comprising a therapeutically effective amount of rhTPP1; use of rhTPP1 for manufacturing an agent for preventing one or more clinical symptoms of CLN2 disease in an individual, or rhTPP1 for preventing one or more clinical symptoms of CLN2 disease in an individual, optionally wherein said individual has a family history of CLN2 disease.

[0060] The present invention further provides a method for treating CLN2 disease, comprising administering to an individual in need a dose of rhTPP1 that effectively maintains or slows or reduces the decline in physiological function, wherein the physiological function is language function, motor function, vision, or eating function. The present invention also provides the use of rhTPP1 in the manufacture of a medicament for maintaining or slowing or reducing the decline in physiological function in an individual suffering from CLN2; and the use of rhTPP1 for maintaining or slowing or reducing the decline in physiological function in an individual suffering from CLN2 disease; wherein the physiological function is language function, motor function, vision, or eating function.

[0061] In one aspect, a method of treating individuals with CLN2 disease or a family history of CLN2 disease involves administering to the individual a dose of rhTPP1 that effectively maintains or slows or reduces language function decline. In one aspect, language function decline is defined as a reduction of at least one point compared to a prior rating determined before or during treatment, as measured using the WCMC or Hamburger Disease Rating Scale. On the WCMC and Hamburger Scales, a rating of 3 indicates normal language; 2 indicates (identifiable) language abnormality; 1 indicates language that is barely intelligible / difficult to understand; and 0 indicates incomprehensible or no language. In one aspect, the dose of rhTPP1 effectively maintains the individual's language rating at the same level as the prior rating determined before or during treatment, such as 3, 2, or 1. In another aspect, the dose of rhTPP1 effectively slows or reduces CLN2-related language function decline in the individual, which can be demonstrated by maintaining the language rating at the same level for a longer period or by a smaller decline in the language function rating compared to what is expected considering the natural progression of the disease.

[0062] In another aspect, treatment of individuals with CLN2 or a family history of CLN2 involves administering an individual a dose of rhTPP1 that effectively maintains motor function or slows or reduces motor function decline. In one aspect, motor function decline is defined as a reduction of at least one point compared to a prior rating determined before or during treatment, as measured using the WCMC or Hamburger Disease Rating Scale. Both the gait scale in the WCMC and the clinical rating scale for movement in the Hamburger scale can be used to assess motor function. In the WCMC and Hamburger scales, a rating of 3 indicates normal walking; 2 indicates abnormal but independent walking, such as frequent falls or marked clumsiness; 1 indicates abnormal walking requiring assistance, such as not walking independently or only crawling; and 0 indicates an individual unable to walk / move, such as being mostly bedridden. In one aspect, a dose of rhTPP1 effectively maintains an individual's motor function rating at the same level as a prior rating determined before or during treatment, such as 3, 2, or 1. In another aspect, the dose of rhTPP1 effectively slowed or reduced CLN2-related motor function decline in individuals, as evidenced by maintaining motor ratings at the same level for a longer period or experiencing smaller declines in motor function ratings compared to what is expected considering the natural progression of the disease.

[0063] In another aspect, a method of treating individuals with CLN2 or a family history of CLN2 involves administering to the individual a dose of rhTPP1 that effectively maintains vision or slows or reduces vision deterioration. In one aspect, vision deterioration is defined as a reduction of at least one point compared to a prior rating determined before or during treatment, as measured using the Hamburger Disease Rating Scale. According to the Hamburger Scale, a rating of 3 indicates that an individual recognizes and grasps a desired object; 2 indicates incoordination in grasping an object; 1 indicates that the individual responds to light; and 0 indicates that the individual does not respond to visual stimuli. In one aspect, the dose of rhTPP1 effectively maintains the individual's vision rating at the same level as the prior rating determined before or during treatment, such as 3, 2, or 1. In another aspect, the dose of rhTPP1 effectively slows or reduces CLN2-related vision deterioration in the individual, which can be demonstrated by maintaining the vision rating at the same level for a longer period of time or by a smaller decline in the vision rating compared to what is expected considering the natural progression of the disease.

[0064] In another aspect, a method of treating individuals with CLN2 or a family history of CLN2 involves administering to the individual a dose of rhTPP1 that effectively maintains or slows or reduces the decline in eating function. In one aspect, the decline in eating function is defined as a reduction of at least one point compared to a previous rating determined before or during treatment, as measured using the WCMC rating scale. According to the WCMC scale, a rating of 3 indicates no swallowing dysfunction; 2 indicates mild swallowing dysfunction; 1 indicates moderate swallowing dysfunction; and 0 indicates dependence on a feeding tube. In this aspect, the dose of rhTPP1 effectively maintains the individual's eating function rating at the same level as the previous rating determined before or during treatment, for example, a rating of 3, 2, or 1. In another aspect, the dose of rhTPP1 effectively slows or reduces CLN2-related eating function decline in the individual, which can be demonstrated by maintaining the eating rating at the same level for a longer period or by a smaller decline in the eating function rating compared to what is expected considering the natural progression of the disease.

[0065] The present invention further provides a method for treating CLN2 disease, comprising administering to an individual in need a dose of rhTPP1 that effectively improves physiological function, wherein the physiological function is language function, motor function, vision, or eating function. The present invention also provides the use of rhTPP1 in the manufacture of a pharmaceutical agent for improving physiological function in individuals with CLN2; and rhTPP1 for improving physiological function in individuals with CLN2; wherein the physiological function is language function, motor function, vision, or eating function. Given the progressive degenerative nature of the disease, improvements in language function, motor function, vision, and / or eating function are particularly needed, indicating that the individual has regained lost functions that are difficult to achieve with current treatment options.

[0066] In one aspect, the method of treating an individual with CLN2 disease includes administering to the individual a dose of rhTPP1 that effectively improves language function. In one aspect, improvement in language function is defined as an increase of at least one point compared to a previous rating determined before or during treatment, as measured using the WCMC or Hamburger Disease Rating Scale. For example, an individual may improve from a rating of 1 or 2 to a rating of 3, indicating a return to normal language function, or from a rating of 1 to a rating of 2.

[0067] In another aspect, methods of treating individuals with CLN2 disease include administering to the individual a dose of rhTPP1 that effectively improves motor function. In one aspect, improvement in motor function is defined as an increase of at least one point compared to a previous rating determined before or during treatment, as measured using the WCMC or Hamburger Disease Rating Scale. For example, an individual might improve from a rating of 1 or 2 to a rating of 3, indicating a return to normal walking, or from a rating of 1 to a rating of 2.

[0068] In one aspect, a method of treating an individual with CLN2 disease includes administering to the individual a dose of rhTPP1 that effectively improves visual acuity. In one aspect, visual acuity improvement is defined as an increase of at least one point compared to a previous rating determined before or during treatment, as measured using the WCMC or Hamburger Disease Rating Scale. For example, an individual may improve from a rating of 1 or 2 to a rating of 3, or from a rating of 1 to a rating of 2.

[0069] In another aspect, methods of treating individuals with CLN2 disease include administering to the individual a dose of rhTPP1 that effectively improves eating function. In one aspect, improvement in eating function is defined as an increase of at least one point compared to a previous rating determined before or during treatment, as measured using the WCMC disease rating scale. For example, an individual may improve from a rating of 1 or 2 to a rating of 3, indicating a return to normal swallowing, or from a rating of 1 to a rating of 2 or 3.

[0070] The present invention further provides a method for treating CLN2 disease, comprising administering to an individual in need a dose of rhTPP1 effective in preventing or treating neurological symptoms of said disease, wherein said neurological symptoms are seizures, decreased brain volume, decreased brain gray matter, or increased cerebrospinal fluid (CSF). The present invention also provides the use of rhTPP1 in manufacturing for the prevention or treatment of neurological symptoms in individuals with CLN2 or a family history of CLN2, and rhTPP1 for the prevention or treatment of neurological symptoms in individuals with CLN2 or a family history of CLN2, wherein said neurological symptoms are seizures, decreased brain volume, decreased brain gray matter, or increased CSF.

[0071] In one aspect, a method of treating individuals with CLN2 or a family history of CLN2 includes administering an individual a dose of rhTPP1 that effectively maintains the frequency of seizures. In one aspect, the dose effectively reduces the number of seizures experienced by the individual each month. In another aspect, as measured using the Hamburger Disease Rating Scale, the dose effectively increases the seizure rating by at least one point compared to a prior rating determined before or during treatment. According to the Hamburger Scale, a rating of 3 indicates no seizures in 3 months; 2 indicates 1 to 2 seizures in 3 months; 1 indicates 1 seizure per month; and 0 indicates more than 1 seizure per month. In one aspect, the dose of rhTPP1 effectively maintains the individual's seizure rating at the same level as the prior rating determined before or during treatment, such as 3, 2, or 1. In another aspect, the dose of rhTPP1 effectively maintains or reduces the frequency of seizures in an individual, which can be demonstrated by maintaining the monthly seizure frequency at the same level for a longer period of time or by a smaller decrease in the seizure rating compared to what is expected considering the natural progression of the disease.

[0072] In another aspect, treatment for individuals with CLN2 or a family history of CLN2 involves administering an individual a dose of rhTPP1 that effectively maintains or slows or reduces the decline in brain volume. Brain atrophy increases with disease progression, causing loss of brain volume and a corresponding increase in volume and intracranial CSF relative to brain volume. Brain volume can be measured using methods known in the art, including imaging techniques such as magnetic resonance imaging (MRI), computed tomography / catalysis (CT / CAT), positron emission tomography (PET), single-photon emission computed tomography (SPECT), electroencephalography (EEG), magnetoencephalography (MEG), and near-infrared spectroscopy (NIRS). In one aspect, a dose of rhTPP1 effectively slows or reduces CLN2-related brain volume decline in an individual, as demonstrated by a longer period of maintained brain volume or a smaller decline in brain volume compared to what is expected considering the natural progression of the disease.

[0073] In another aspect, methods for treating individuals with CLN2 or a family history of CLN2 involve administering to the individual a dose of rhTPP1 that effectively maintains or slows or reduces the decline in gray matter in the brain. Gray matter loss due to brain atrophy occurs as the disease progresses, resulting in a decrease in gray matter as a percentage of brain volume. The amount of gray matter in the brain can be assessed using methods known in the art, such as imaging techniques like MRI, CT / CAT, PET, SPECT, EEG, MEG, and NIRS. In one aspect, a dose of rhTPP1 effectively slows or reduces the decline in gray matter in an individual, which can be demonstrated by maintaining gray matter volume for a longer period or by a smaller decrease in gray matter as a percentage of brain volume compared to what is expected considering the natural progression of the disease.

[0074] In another aspect, methods for treating patients with CLN2 or a family history of CLN2 involve administering to the individual a dose of rhTPP1 that effectively maintains or slows the increase in cranial CSF volume. Due to brain atrophy, cranial CSF increases in both volume and proportion of the total CSF. The amount and proportion of cranial CSF can be assessed using methods known in the art, such as imaging techniques like MRI and CT / CAT. In one aspect, a dose of rhTPP1 effectively slows or reduces the increase in cranial CSF in an individual, which can be demonstrated by maintaining cranial CSF volume for a longer period or by a smaller increase in cranial CSF as a percentage of the total CSF compared to what is expected considering the natural progression of the disease.

[0075] The above methods, compositions, and uses may further include any of the following features, individually or in combination.

[0076] In one aspect, the method, use of the composition, or application of the present invention comprises continuously or intermittently administering to an individual a formulation, composition, or dose containing rhTPP1 for a period of at least about 1 hour, for example, at least about 1 hour, at least about 2 hours, at least about 3 hours, at least about 4 hours, at least about 5 hours, at least about 6 hours, or more. In another aspect, the method or application of the present invention comprises administering to an individual in need a formulation, composition, or dose comprising about 20 mg to about 500 mg, about 30 mg to about 500 mg, about 50 mg to about 500 mg, about 100 mg to about 500 mg, about 200 mg to about 400 mg, about 250 mg to about 350 mg, or about 275 mg to about 325 mg of rhTPP1, for example, about 20 mg, about 30 mg, about 50 mg, about 100 mg, about 200 mg, about 300 mg, about 400 mg, or about 500 mg of rhTPP1. In one aspect, the method or use of the present invention comprises administering a formulation, composition or dose having a volume of about 20 mL or less, about 15 mL or less, about 10 mL or less, about 7.5 mL or less, or about 5 mL or less per dose or per dosing event, such as about 20 mL, about 15 mL, about 10 mL, about 9 mL, about 8 mL, about 7 mL, about 6 mL, about 5 mL, about 4 mL, about 3 mL, about 2 mL, about 1 mL, or about 0.5 mL.

[0077] In various aspects, the methods, uses, or applications of the present invention comprise administering to an individual a formulation, composition, or dose comprising rhTPP1 at a rate of less than or equal to about 2.5 mL of the formulation, composition, or dose per hour, less than or equal to about 75 mg rhTPP1 per hour, or less than or equal to about 75 mg rhTPP1 per 2.5 mL of the formulation or composition per hour. The formulation, composition, or dose may optionally be administered continuously or intermittently over a period of at least about 4 hours.

[0078] In one aspect, the method, use of the composition, or application of the present invention comprises administering a formulation, composition, or dose comprising rhTPP1 weekly or less frequently, such as weekly, every other week, or monthly. More specifically, the method, use of the composition, or application of the present invention comprises administering a formulation, composition, or dose comprising rhTPP1 once every 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, or 31 days. In one aspect, the formulation, composition, or dose is administered intraventricularly. In another aspect, the formulation, composition, or dose is administered intrathecally. In yet another aspect, the formulation, composition, or dose is administered intraocularly. In one aspect, the formulation, composition, or dose is administered intraventricularly, intrathecally, and intraocularly. Attributable to the physiology of intraventricular delivery, which allows CSF flow into the third and fourth ventricles, intraventricular delivery allows penetration into the brain's dark gray structures, such as the thalamus, striatum, and midbrain, and also across the neural fiber network of the cerebral hemispheres along a slight pressure gradient from the ventricles to the subarachnoid space. Intrathecal and intraventricular administration of recombinant enzymes for the treatment of lysosomal storage disorders is described in U.S. Patent No. 7,442,372, which is incorporated herein by reference in its entirety.

[0079] The formulations, compositions, or doses of rhTPP1 of the present invention can be administered by a single rapid injection or a series of injections (e.g., into the brain, waist, or eye), or as a continuous or sustained infusion, such as using an infusion pump or other implantable device. In one aspect, the formulations, compositions, or doses of rhTPP1 are administered using an infusion system comprising a tank, an in-line filter (e.g., about 0.2 μm), a reservoir (e.g., intrathecal or intraventricular), and a catheter. Often, when a composition is administered intrathecally or intraventricularly, to prevent adverse side effects caused by artificially increasing intracranial or intrathecal pressure, a volume of CSF equivalent to the volume of the composition to be administered is removed from the individual before administration. As described in Example 3, however, it is demonstrated herein that the formulations, compositions, or doses of rhTPP1 of the present invention can be administered without removing any volume of CSF from the individual before administration of the rhTPP1 formulation, composition, or dose.

[0080] In one aspect, the method or use of the present invention includes administering approximately 10 mL of a formulation, composition, or dose comprising approximately 300 mg rhTPP1 to an individual with CLN2 every other week over a period of approximately 4 hours.

[0081] The formulations and compositions of the present invention can be administered directly to individuals in need (i.e., non-isovolume) or after the removal of a defined volume of CSF from the individual, wherein the defined volume is substantially the same as the volume of the composition subsequently administered (i.e. isovolume).

[0082] In one aspect, the method, use of the composition, or application of the present invention further comprises administering a flushing solution to an individual after administration of rhTPP1. The flushing solution is administered via the same route as rhTPP1 and using the same delivery system (e.g., infusion system) to remove any rhTPP1 remaining in the delivery system and to ensure that the individual receives the full intended dose of rhTPP1. In one aspect, the flushing solution is administered to the individual in an amount between about 0.5 mL and about 5 mL, such as about 0.5 mL, about 1 mL, about 2 mL, about 3 mL, or about 5 mL (e.g., using the same catheter previously used to administer compositions comprising rhTPP1). In one aspect, the flushing solution comprises the same components as a formulation or composition comprising rhTPP1, but without rhTPP1. In one aspect, the rinsing solution comprises about 0.11 mg / mL of disodium hydrogen phosphate heptahydrate, about 0.08 mg / mL of sodium dihydrogen phosphate monohydrate, about 8.77 mg / mL of sodium chloride, about 0.22 mg / mL of potassium chloride, about 0.16 mg / mL of magnesium chloride hexahydrate, about 0.21 mg / mL of calcium chloride dihydrate, and a diluent, such as water for injection.

[0083] Reagent test kit

[0084] The present invention further provides a kit comprising a formulation of rhTPP1 as described herein, said formulation being in a dosage and form suitable for administration to a patient. In one aspect, the kit comprises about 30 mg / mL of rhTPP1, about 0.11 mg / mL of disodium hydrogen phosphate heptahydrate, about 0.08 mg / mL of sodium dihydrogen phosphate monohydrate, about 8.77 mg / mL of sodium chloride, about 0.22 mg / mL of potassium chloride, about 0.16 mg / mL of magnesium chloride hexahydrate, about 0.21 mg / mL of calcium chloride dihydrate, and a diluent, such as water for injection. In one aspect, in addition to the therapeutic formulation, the kit further comprises instructions for intraventricular, intrathecal, and / or intraocular administration of the therapeutic compositions of the present invention. In another aspect, the kit further comprises an irrigation solution as described herein. In yet another aspect, the kit further comprises a system for administering the formulation, said system comprising any one or all of the following components: a tank, an in-line filter, a reservoir for implantation, and a catheter. In one aspect, the kit may include a catheter, reservoir, or other device preloaded with the therapeutic formulation of the present invention. For example, this specifically includes catheters preloaded with about 100 mg, about 200 mg, about 300 mg, about 400 mg, or about 500 mg of rhTPP1 in a pharmaceutically acceptable formulation. Alternatively, the kit may include a refillable catheter, reservoir, or other device and an appropriate amount of enzyme for refilling such devices.

[0085] In some embodiments, the kit of the present invention may include one or more of the following components: an extension tubing (e.g., product number 536040, Smiths Medical, Dublin OH), an in-line filter (e.g., product number FS116, Smiths Medical), a port needle (e.g., product number 21-2737-24, Smiths Medical), a syringe or two or more syringes (e.g., product number 309604, Becton Dickinson, Franklin Lakes, NJ) or syringe needles or two or more syringe needles (e.g., product number 305196, Becton Dickinson).

[0086] The invention will be more readily understood by referring to the following examples, which are provided by way of illustration and are not intended to be limiting.

[0087] Example

[0088] The following examples describe the results of administering the formulation containing rhTPP1 to intraventricular (ICV) patients and comparing the results of administering the formulation to human patients with matched untreated natural history patients.

[0089] Example 1

[0090] formulations of rhTPP1 for intraventricular administration

[0091] RhTPP1 was produced in genetically engineered CHO host cell lines and purified by standard chromatography, as described in U.S. Patent No. 6,302,685, which is incorporated herein by reference in its entirety, and in Sleat et al., 1997, Science 277:1802-1805. rhTPP1 is regenerated as an inactive proenzyme upon uptake to lysosomes at acidic pH. The proenzymatic form of rhTPP1 has a calculated isotopic average molecular weight of approximately 59 kDa. The mature enzyme has an apparent molecular weight of approximately 46 kDa. The amino acid sequence of the rhTPP1 proenzyme is illustrated in SEQ ID NO:1 and shown in [image / image / contact information]. Figure 1 The pre-enzyme segment consists of the first 176 amino acid residues, while the mature enzyme is 368 amino acids long, starting from position 177, and is described in SEQ ID NO:2.

[0092] The rhTPP1 formulation used in this example is a sterile solution for ICV infusion. It is a clear, colorless to pale yellow liquid containing rhTPP1 protein at a concentration of 30 mg / mL. The formulation is packaged in a container closure system consisting of Type 1 clear borosilicate glass vials, which are sealed with fluoropolymer-coated butyl rubber stoppers and capped with aluminum. The formulation is stored at -40°C ± 10°C and supplied frozen. The target pH of the formulation is pH 6.5.

[0093] The compositions used for the rhTPP1 formulations in the examples are provided in Table 2.

[0094] Table 2

[0095]

[0096] The rhTPP1 formulation was carefully designed to mimic the characteristics of human CSF, such as the concentrations of key electrolytes being similar to those found in human CSF in vivo, and the formulation contains no conventional preservatives or stabilizers as excipients. No significant safety issues, i.e., serious adverse reactions, that might not have been previously predicted, have been reported or observed after administration of the rhTPP1 formulation.

[0097] In accordance with ICH guidelines and protocols, stability studies were conducted under long-term (≤-60°C) and accelerated conditions (5±3°C) to monitor time-temperature stability. Stability samples were stored in small-scale vials made of the same materials as the large-scale packaging. Stability data collected from supportive and clinical batches demonstrated that the rhTPP1 formulation was stable for at least 36 months at ≤-60°C and at least 6 months at 5±3°C, which is surprising considering the absence of preservatives and stabilizers typically found in traditional Chinese medicine products. Table 3 presents the results of the stability tests.

[0098] Table 3

[0099]

[0100] Example 2

[0101] Natural History Studies

[0102] A quantitative assessment of the natural history of CLN2 disease progression was analyzed in a cohort of 41 untreated CLN2 patients. The Hamburger Clinical Scale was used to assess age-appropriate neurological and functional domains of disease.

[0103] A quantitative description of clinical decline in untreated natural history CLN2 individuals is presented in Figure 2 Natural history analysis demonstrates a transparent and predictable relationship between age and disease severity. Following the onset of motor and language symptoms, a generally rapid linear decline occurs, with the average child losing approximately two milestone events per year (a linear rate of decline of 2.1 points per year). The process is generally predictable; however, there are some 'late-onset' cases, comprising less than 20% of the population. These patients typically have late-onset symptoms and a longer duration of mild illness, followed by rapid and active decline, usually 2 to 3 years later than the conventional form.

[0104] Quantitative clinical progression from the Hamburg group was assessed by aggregating clinical ratings from an independent (patient and ratingr) cohort (n=49) from the WCMC. While the clinical descriptions of the independent CLN2 groups were similar, this was the first confirmation of a strong quantitative relationship in disease progression in another patient group. Both CLN2 patients had most conventionally delayed onset and progression in infancy, with a smaller proportion of children exhibiting a 'late onset' phenotype, typically presenting with early symptoms at age 5 rather than 3. Neurological decline in CLN2 patients was reproducibly captured using scales for motor (gait) and language function. Based on the above analysis, the natural history group was determined to be an appropriate untreated control population, and the mean rate of decline of CLN2 disease symptoms in this untreated natural history population can serve as an effective and informative comparative object for any prevention or reduction in the rate of decline of symptoms induced by administration of the compositions of the present invention in individuals with CLN2 disease.

[0105] Example 3

[0106] Phase 1 / Phase 2 Open Label Dose Expansion Study in CLN2 Patients

[0107] This study was an open-label clinical trial evaluating the safety, tolerability, and efficacy of the rhTPP1 formulation of this invention, delivered weekly to children with CLN2 disease via an ICV catheter at a dose of 300 mg (10 mL total volume). The study was designed to assess safety and tolerability, starting with low doses (30 mg and 100 mg), but all patients were escalated to a high expected treatment dose (300 mg) when the independent data monitoring committee noted the safety of the lower doses. The study duration for all enrolled patients was 48 weeks of treatment with a stable expected treatment dose of 300 mg every other week via ICV. The primary objective was to evaluate the safety and tolerability of the rhTPP1 formulation of this invention administered via an implanted ICV reservoir and cannula to individuals with CLN2 disease and to assess its validity compared to natural history data using CLN2 disease-specific rating scales after 12 months of treatment. A secondary objective was to assess the effect of treatment on measurements of brain atrophy after 12 months of treatment, compared to natural history data for CLN2 disease.

[0108] The primary inclusion criteria were a CLN2 diagnosis and an age of at least 3 years. Patients with a baseline disease rating score less than 3 at screening (using the Hamburger 0–6 Total Motor / Verbal Scale) were excluded from the study. Patients younger than 3 years were more likely to be non-progressive due to age rather than treatment, as depicted by the horizontal line on the progression curve. Patients with a score of 2 or lower at screening were attributed to disease stage, which is more nonlinear, more variable, and potentially more difficult to treat. Therefore, the treatment group was defined solely by age and score to include early and highly predictable decline.

[0109] The mean age at enrollment was 4.0 years, with slightly more girls than boys, and the majority were Caucasian. Clinical CLN2 scores at screening and baseline are shown in Table 4 below, which displays the Hamburger Motor / Verbal Scores and total scores at screening and baseline for each study group.

[0110] Table 4

[0111]

[0112] In summary, pre-treatment CLN2 scores tended to favor later-stage disease. Assuming rapid disease progression and difficulty in identification, a bias towards lower scores is expected. Furthermore, some score decline occurred during the period from screening to baseline assessment (up to two weeks before ICV reservoir placement). Four patients with a score of 3 from the screening group slipped one point from baseline, and two patients with a score of 4 from the screening group lost one point from baseline to 3. The two patients who entered with a score of 6 (i.e., substantially normal) were siblings of the affected child. The distribution, demographics, and characteristics of the individual populations are summarized in Table 5 below.

[0113] Table 5

[0114]

[0115]

[0116] a Patients 1287-1007 who received a single dose and agreed to withdraw due to failure to meet study procedures were enrolled.

[0117] b Similar to the distribution of natural history groups

[0118] * Common genotypes: c.622C>T and c.509-1G>C

[0119] All enrolled patients received a steady-state dose of 300 mg ICV every other week. Group 1 patients were exposed to 30 mg ICV every other week for ≥1 month, then increased to 100 mg ICV every other week for ≥4 weeks, while Group 2 patients started with 100 mg ICV every other week for ≥4 weeks. Groups 1 and 2 patients increased to 300 mg ICV every other week, and all subsequent patients, including those in Group 3, started a steady-state dose of 300 mg ICV every other week for ≥48 weeks. The 300 mg dose was administered in 10 mL via the ICV catheter over approximately 4 hours. Just before starting the infusion, a volume of CSF equal to the amount of rhTPP1 formulation administered was not removed; this was atypical but unexpectedly did not cause any side effects. Immediately after administering the 300 mg dose, approximately 2 mL of flushing solution was administered to the individual via the same ICV catheter. The flushing solution was consistent with the formulations in Table 2, but without rhTPP1. The 300 mg bolus dose of enzyme per dosing event was significantly higher than previous intrathecal or ICV-administered enzyme replacement therapy, thus the safety and efficacy profiles observed after administration of such high doses could not have been predicted previously. More specifically, the absence of associated serious, difficult-to-manage adverse events with a 300 mg hTPP1 bolus dose may not have been previously predicted.

[0120] result

[0121] The role of treatment in the clinical assessment of gait and language: The primary assessment tool for quantitative evaluation of clinical severity is the 0-6 total of the gait and language subscales common to the Hamburger and WCMC Disease Rating Scales. This scale captures predictable, rapid, and progressive clinical decline in matched, untreated, natural-history patients used as the comparison item in the primary efficacy analysis.

[0122] Gait / language disorder rating scores of 23 patients whose treatment duration exceeded 42 weeks are displayed on Figures 3A to 3F Of the 23 patients, 3 (1244-1001, 1244-1002, and 1244-1003) were from group 1 (C1), 3 (1244-1004, 1244-1006, and 1247-1005) were from group 2 (C2), 3 (1244-1008, 1244-1009, and 1244-1010) were from group 3 (C3), and 14 (0119-10) were from group 4 (C1). 20, 0146-1021, 0146-1022, 0146-1023, 1244-1011, 1244-1012, 1244-1017, 1244-1024, 1323-1013, 1323-1014, 1323-1015, 1323-1016, 1323-1018, and 1323-1019) were from the 300 mg stable-dose (SBO) group only. As expected, language impairment usually occurred later than gait impairment. The inclusion scores were not randomly distributed; 12 patients had significant disease progression with a combined inclusion score of 3, and 2 patients had a combined inclusion score of 6. Assuming rapid progression and disease confirmation, children often exhibit significant regression or resemble siblings of those with significant regression.

[0123] After treatment with the rhTPP1 formulation of the present invention (shown in Table 2 above), the CLN2 gait / language disorder score stabilized, as shown in Table 2 above. Figures 3A to 3F As shown in the figure. Eleven of the 23 patients experienced a reversal of their rating decline during the treatment period. Four patients had a single-unit decline at the beginning of the treatment period, but the decline was subsequently reversed. Two patients (1244-1008 and 1323-1013) experienced a one-unit decline from 3 to 2 between screening and baseline, but did not experience any other loss in rating during treatment. Based on these results, there was a significant treatment benefit in all patients, regardless of the group (starting dose) or the inclusion score. Reversed rating declines were observed in a large number of patients. For example, patients 1287-1005 ( Figure 3B The patient exhibited a 2-unit reduction in the rating during the first month of treatment, indicating loss of gait and language function. However, this patient regained one unit on day 60 of treatment and remained unchanged thereafter until day 440 of the analysis. The regained rating represents language acquisition, highlighting the clinical significance of a single unit change.

[0124] Neither of the two patients with a score of 6 at enrollment had lost a rating unit. Of the 12 patients with a score of 3 at enrollment, seven experienced a reversal of decline, and two stabilized after an initial decline of one unit. Therefore, the treatment benefit was evident in patients with significant deficiencies and disease progression.

[0125] As demonstrated in the CLN2 natural history study, the median rate of decline in the untreated natural history population was estimated at 2.1 units per year. Therefore, all patients in the treatment group had improved ratings compared to the untreated natural history population.

[0126] To establish a clearer relationship between treated and matched untreated natural history patients during the course of the disease, study patients were matched with untreated natural history patients using parameters such as baseline CLN2 score, age, and genotype. While no clear subgroup or factor predicts progression in CLN2 disease, these parameters are most commonly used to define disease severity. Individual treated patients were compared to members of the natural history group with similar gait / language ratings at baseline, such as... Figures 4A to 4I As shown in the diagram. Patients in the study were matched by baseline CLN2 scores as follows: For a given study patient with a given baseline score, all natural history patients with reported one or more CLN2 assessments having the same CLN2 score were identified. If the study patient's baseline CLN2 score was 2, 3, 4, or 5, then the CLN2 time morphology of each natural history patient was shifted left or right temporally to cover the study patient's baseline score. If a natural history patient had multiple assessments equal to the study patient's baseline CLN2 score, then the intermediate time point of the multiple assessments was used for time shifting. If the study patient's baseline CLN2 score was 6, then the natural history patient's previous score of 6 was used for time shifting. Sensitivity analyses were performed using other matching criteria, and the results from these analyses were consistent with the analysis of score matching.

[0127] Figures 4A to 4I The results of mapping individuals treated with the rhTPP1 formulation of this invention relative to matched untreated natural history patients are presented. Treated individuals and untreated natural history patients were matched using disease rating scales, with gait and language subscales from 0 to 6 units as the total. Each rating was compared over a one-year treatment period. A treatment benefit was observed in individuals receiving rhTPP1 compared to the matched untreated natural history patient group. Individuals 1244-1001 ( Figure 4A The patient experienced a decline in grade from 3 units to 2 units after 120 days of treatment, but later regained one unit with no net change. Individual 1244-1002 Figure 4BThe study showed increases in ratings from 3 to 4 units, decreases from 4 to 2 units, and increases from 2 to 3 units, resulting in an overall maintenance of disease ratings compared to day 1 at the end of the study. Individuals 1244-1003 ( Figure 4C ) and 1244-1010 ( Figure 4I Throughout the study period, the individual maintained a rating of 6 units, indicating normal motor and language function. Individual 1244-1004 Figure 4D ) and 1244-1009 ( Figure 4H The rating was maintained at 3 units throughout the study. Individuals 1244-1006 ( Figure 4E The initial rating decreased from 3 units to 2 units, but then regained one unit with no net change, before decreasing again from 3 units to 2 units. Individual 1244-1008 Figure 4G The rating initially declined from 3 units to 2 units, but then remained unchanged.

[0128] Compared to all treated individuals, the majority of their matched untreated natural history patients had irreversible grade declines ranging from 3 to 0 units by the end of the comparison period, indicating progression to complete loss of combined gait and language function. Matched analysis demonstrated treatment benefit for these patients who maintained their disease grade scores and those with initial grade declines that subsequently stabilized.

[0129] The most complex response (individuals 1287-1005) is shown in Figure 4F Although this patient experienced a rapid decline of 2 units from a baseline score of 3 to a study score of 1 in the first month of the study, the patient was able to regain one unit and stabilize at a score of 2. The explanation for this process was elucidated by comparison with score-matched untreated natural history patients. In 15 of the 18 score-matched untreated natural history patients, clinical progression was worse, and the same was true in only 2 score-matched untreated natural history patients (a single untreated natural history match is not estimable). Clinical course consistently worsened in untreated patients, with shorter intervals between lost milestones. Lost function was never re-established and subsequently stabilized. Therefore, matching with the most complex treatment patterns also demonstrated a clear treatment benefit.

[0130] Figure 5This shows the distribution of clinical changes from baseline in matched untreated natural history patients compared to the study individuals during the patient-matched treatment duration. As mentioned above, 7 out of 9 patients (>75%) showed no change in baseline disease rating scales. For those 7 patients during the treatment period, all matched untreated natural history patients had at least a single unit decline, but more commonly multiple unit declines or 2 to 4 units. For example, patients 1244-1001 had 1 match with a single point loss, 3 matches with a 2-point loss, and 14 matches with all 3 available language / gait disease rating points lost. Therefore, during the same period, the treated patients showed no change, but 14 out of 18 matched untreated natural history patients (>75%) lost all gait and language functions. A base effect was observed at the baseline inclusion score of 3, with many matched untreated natural history patients losing all available rating units. However, two patients with inclusion scores of 6 (patients 1244-1003 and 1244-1010) significantly demonstrated that these matches also actively regressed, with some showing a decline to 4 and 5 points during the treatment period. This observation provides clear clinical evidence of the substantial therapeutic effect; most treated children retained their inclusion clinical ratings against the backdrop of active loss of language and independent gait in the untreated natural history matches during the same period, with many showing complete functional loss. The remaining two treated patients who lost single points (individuals 1287-1005 and 1287-1006) remained better than most matched clinical ratings during the treatment period. Overall, 97% of the score-matched untreated natural history patients had worse ratings than the treated individuals.

[0131] Using multiple matching criteria (e.g., baseline and genotype), nearly 100% of comparisons showed favorable treatment outcomes compared to matched untreated natural history patients. The mean treatment difference across matching criteria ranged from 1.9 to 2.1 points across all treated patients compared to natural history patients, depending on the matching criteria used.

[0132] Figure 9A The mean change in motor-verbal ratings is shown for patients treated for ≥48 weeks (n=21; dashed line) and the natural history group (n=41; solid line). The mean decline in disease ratings for treated patients was 0.43 (standard deviation 0.839), with a median decline of 0.00 units over 48 weeks. In contrast, the mean decline in disease ratings for the natural history group was 2.09 (standard deviation 0.966), with a median decline of 1.87 units over 48 weeks. Overall, 79% of the expected clinical decline was significantly preserved in treated patients (p<0.0001). Figure 9BThis study presents the changes in motor-verbal scores in patients (n=23) from the last measurement taken ≥48 weeks prior to the first 300 mg dose (baseline). Overall, 65% (15 of 23) of patients experienced improvement or no clinical disease progression during treatment, and 87% (20 of 23) of patients demonstrated better efficacy (i.e., a score change of -1 or greater) during treatment compared to treatment-naïve individuals from the natural history study. These analyses unanimously support the conclusion that significant and clinically meaningful stabilization of the CLN2 score was observed in treated patients compared to matched members of the treatment-naïve natural history group, who experienced rapid and predictable decline.

[0133] The role of treatment in the clinical assessment of vision: In untreated CLN2 patients, vision loss occurs later than language and gait decline; however, once symptoms appear, the process is predictably rapid and progresses to blindness. Therefore, protecting vision is an important treatment outcome. Vision loss can be captured using a 0-3 point subscale, similar to other subscales, where 3 represents normal and 0 represents functional blindness. Most treated patients experienced reversal of loss in the vision subscale domain during the treatment period. When treated patients were matched with untreated natural history individuals using a combined scale (0-9 units) of the gait, language, and vision subscales based on scores, age, and genotype, it was clear that the untreated matched natural history individuals had additional points lost compared to the treatment group.

[0134] Figures 6A to 6I The results of mapping nine individuals treated with the rhTPP1 formulation of this invention relative to matched untreated natural history patients are presented. Patients were matched using disease rating scales, with gait, language, and visual acuity subscales ranging from 0 to 9 units as the total. Individuals 1244-1001 ( Figure 6A The individual experienced a one-point drop in rating from 6 to 5, but soon regained one point to a rating of 6, after which there was no net change. Individual 1244-1002 Figure 6B The disease rating increased from 5 to 6 points, then decreased from 6 to 4 points, and then increased from 4 to 5 points, resulting in an overall stable disease rating compared to day 1 at the end of the study. Individuals 1244-1003 ( Figure 6C ) and 1244-1010 ( Figure 6I A score of 9 was maintained throughout the study, indicating normal gait, language function, and vision; individual scores ranged from 1244 to 1004. Figure 6D ) and 1244-1009 ( Figure 6H The score was maintained at 6 throughout the study, and individuals 1244-1008 ( Figure 6G The score was maintained at 5 throughout the study. Individuals 1244-1006 Figure 6EAn initial rating of 6 to 5 points decreased by one unit, then further decreased to 4 points, but regained a rating of 5 points by one unit, after which there was no net change. Individual 1287-1005 Figure 6F It has a rating that drops from 6 to 4, then increases from 4 to 5 and drops back to 4, but then regains a unit to the final rating of 5.

[0135] The inclusion of the visual acuity subscale did not cause changes in the nine treated patients during the treatment period. However, the effect of matching untreated natural history patients on visual acuity loss to the total score was significant. Several untreated natural history matches differed from treated patients by more than 3 points, demonstrating the impact of visual acuity decline on the total score during the study period. Therefore, the inclusion of the visual acuity subscale increased the difference between treated patients and matched untreated natural history patients. Because the loss of disease rating units was reversed in matched untreated natural history patients, the observed treatment effects of rhTPP1 related to preventing disease progression and stabilizing function can be extended from motor / gait and language to the clinical domain including visual acuity.

[0136] The role of treatment in overall disease assessment: During the study, patients were also assessed using a combined 12-point scale containing either the complete Hamburger or WCMC scores. The scores on the 12-point scale were the sum of the individual scores for (1) motor / gait, (2) language, (3) tics / myoclonus, and (4) vision / eating. Figures 10A to 10L The results show the outcomes of individuals treated with the rhTPP1 formulation of this invention using 0 to 12 units of the combination hamburger (left panel) and the WCMC scale (right panel). Sixteen of the 23 patients showed reversal of decline on at least one scale, and eight patients showed an increase in scores on at least one scale compared to baseline at the end of the treatment period, confirming the overall treatment benefit in patients receiving rhTPP1.

[0137] Effects of treatment on brain volume: MRI was used to assess secondary endpoints in treated patients. Disease course is characterized by atrophy, cell loss, and signal abnormalities. These parameters, individually or as a composite score, are correlated with patient age and disease rating scores. Therefore, there is general agreement that disease progression is associated with MRI indices of atrophy, and several MRI parameters have been shown to be correlated with age and disease severity in CLN2 disease (Dyke et al., AJNR AmJ Neuroradiol. 2013; 34(4):884-9; Paniagua et al., Clin Neuroradiol. 2013; 23(3):189-96). The imaging databases supporting these conclusions are based on cross-sectional correlations of a significant number of patients; however, longitudinal acquisitions within patients without MRI imaging are not available. Therefore, longitudinal studies derived from MRI analyses cannot be equivalently matched with similar derived natural history databases.

[0138] To analyze treated patients, MRI parameters were standardized across the hardware platform at the study sites. Data was acquired locally, discriminatory information was edited, and then transmitted to the central imaging core laboratory. Imaging was randomized, ensuring that independent radiologists were unaware of the patient's or time's relationship to baseline. Brain volume changes were reconstructed from randomized, independent central readings. Data were analyzed across the treatment population, compared to baseline, and longitudinally. Figure 7 This section presents an overview of brain volume measurements taken by MRI in treated patients. Brain atrophy results in a larger volume and proportion of intracranial CSF. This increase in these measurements is correlated with age and severity in CLN2 patients. Longitudinal curves of CSF volume and proportion in treated patients suggest that any changes in CSF parameter measurements appear to be minimal. All patients had MRI volume analyses that showed constancy and stability consistent with rating assessments.

[0139] Figure 8A to 8L Longitudinal MRI assessment of brain volume in treated patients is presented. Autonomic degeneration in CLN2 patients is characterized by significant gray matter loss and compensatory increase in CSF. However, during the assessment period, very stable brain volume was observed, and there was no evidence of neurodegenerative processes in treated patients. For each treated patient, Figure 8A to 8L The gray matter volume shown in each of the top and bottom plots as the difference between the CSF and gray matter curves (dashed lines) and the CSF curves (solid lines) remained stable throughout the study. The change in cortical gray matter volume as a percentage of total brain volume, compared to the last observation after ≥48 weeks of treatment (baseline) prior to the first 300 mg infusion, is shown in Table 6 below.

[0140] Table 6

[0141]

[0142] The longitudinal change in cortical volume was -1% per year in normal children aged 4-12 years, but -12.5% ​​per year in untreated CLN2 patients. During rhTPP1 treatment, the volumes of CSF, gray matter, and white matter remained relatively constant, reducing disease-related cortical volume loss by 89%.

[0143] Adverse events: One patient withdrew from the study due to protocol non-compliance. The remaining 23 patients remained in the study and were permitted to be treated with the rhTPP1 drug product via the ICV route. There were no deaths, treatment-related withdrawals, or study interruptions due to safety-related reasons. Consistent with the minimal impact of device implantation, all patients were administered medication within one week of the procedure. Of the total 325 infusions, only 5 (1.5%) were interrupted for any reason, and of these, only 2 (0.6%) were interrupted for reasons related to adverse events. The most frequent non-CLN2 disease-related adverse events observed in the study were fever, hypersensitivity reactions, and upper respiratory tract infections (each in 25% of the entire individuals). Generally, these events were mild, self-limiting, and medically managed. Hypersensitivity events, as defined by the investigators, were associated with a small number of peripheral presentations and were medically managed with a combination of antipyretics, antihistamines, and / or steroids. Laboratory data demonstrated a lack of clinically relevant changes in peripheral laboratories. In CSF, some patients had mild, transient cerebrospinal fluid cytosis with no changes in CSF glucose or protein. In summary, the evaluation of safety parameters demonstrates that the rhTPP1 formulation of the present invention is permissible for treatment via ICV infusion in all patients.

[0144] in conclusion

[0145] Clinical studies have demonstrated a significant clinical benefit in every patient with treatment exposure exceeding 36 weeks, characterized by complete suppression of CLN2 disease progression. This constitutes the greatest treatment benefit because no functional increase was expected in patients with moderate progression and active degeneration within this timeframe.

[0146] This finding is even more compelling when patients are matched to members of a natural history database based on multiple parameters including baseline disease rating, age, and genotype. This matching revealed that matched untreated natural history patients experienced substantial functional loss during the same period when treated individuals experienced cessation of disease progression while on rhTPP1 therapy. Therefore, all treated patients showed active disease progression cessation compared to matched untreated natural history patients. Based on available natural history data, the median rate of decline in the untreated natural history population was estimated at 2.1 units per year, where each unit of decline represents a significant milestone loss of physiological function. For most patients enrolled in the study, maintenance of 2 units translated into sustained independent walking and meaningful communication.

[0147] Overall, the results demonstrate that the rhTPP1 formulation and treatment method of this invention have an acceptable safety / tolerability profile. No individuals discontinued the study or treatment due to adverse events. One individual withdrew from the study after a single treatment dose due to protocol noncompliance. PK and immunogenicity analyses revealed high CNS delivery and no antibody formation in CSF.

[0148] The above examples demonstrate that formulations and methods including rhTPP1 described herein are effective in preventing or treating CLN2 disease and / or one or more CLN2 clinical symptoms. In diseases characterized by rapid, relentless, and irreversible neurodegenerative disease progression, halting disease progression, particularly in each treated patient, is a substantial and unexpected clinical benefit.

[0149] Examples of embodiments of the present invention:

[0150] 1. A formulation comprising recombinant human tripeptidyl peptidase-1 (rhTPP1) for intraventricular, intrathecal, or intraocular administration.

[0151] 2. The formulation according to embodiment 1, wherein the rhTPP1 comprises the amino acid sequence of SEQ ID NO:1 or a fragment thereof.

[0152] 3. The formulation according to embodiment 1 or 2, wherein the formulation comprises rhTPP1 at a concentration of about 25 mg / mL to about 35 mg / mL.

[0153] 4. The formulation according to any one of embodiments 1 to 3, wherein the formulation has a pH value of about 6.5.

[0154] 5. The formulation according to any one of embodiments 1 to 4 further comprises potassium chloride at a concentration of about 0.01 mg / mL to about 1 mg / mL, magnesium chloride hexahydrate at a concentration of about 0.01 mg / mL to about 1 mg / mL, and calcium chloride dihydrate at a concentration of about 0.01 mg / mL to about 1 mg / mL.

[0155] 6. The formulation according to any one of embodiments 1 to 5 further comprises a concentration of disodium hydrogen phosphate heptahydrate of about 0.01 mg / mL to about 1 mg / mL, a concentration of sodium dihydrogen phosphate monohydrate of about 0.01 mg / mL to about 1 mg / mL, and a concentration of sodium chloride of about 1 mg / mL to about 20 mg / mL.

[0156] 7. The formulation according to any one of embodiments 1 to 6 comprises about 0.11 mg / mL of disodium hydrogen phosphate heptahydrate, about 0.08 mg / mL of sodium dihydrogen phosphate monohydrate, about 8.77 mg / mL of sodium chloride, about 0.22 mg / mL of potassium chloride, about 0.16 mg / mL of magnesium chloride hexahydrate, and about 0.21 mg / mL of calcium chloride dihydrate.

[0157] 8. The formulation according to any one of embodiments 1 to 7, wherein the formulation does not contain preservatives.

[0158] 9. The formulation according to any one of embodiments 1 to 8, wherein the formulation is stable at about 5°C for at least about 6 months.

[0159] 10. A method for treating neuronal ceroid lipofuscin deposition disease (CLN2), comprising administering to an individual in need a formulation according to any one of embodiments 1 to 9.

[0160] 11. A composition comprising a formulation according to any one of embodiments 1 to 9 for treating neuronal ceroid lipofuscin deposition disease (CLN2).

[0161] 12. Use of a formulation according to any one of embodiments 1 to 9 for manufacturing a medicament for treating neuronal ceroid lipofuscinosis (CLN2).

[0162] 13. A method of treating neuronal ceroid lipofuscinosis (CLN2) disease, comprising administering to an individual in need about 10 mL of a composition comprising a dose of about 300 mg of recombinant human tripeptidyl peptidase-1 (rhTPP1), wherein the composition is administered to the individual over a period of about 4 hours.

[0163] 14. A method of treating neuronal ceroid lipofuscinosis (CLN2) disease, comprising administering to an individual in need a dose of about 300 mg of recombinant human tripeptidyl peptidase-1 (rhTPP1), wherein said dose is administered to said individual at a rate of less than or equal to 75 mg / h.

[0164] 15. A method of treating neuronal ceroid lipofuscinosis (CLN2) disease, comprising administering to an individual in need a dose of about 300 mg of recombinant human tripeptidyl peptidase-1 (rhTPP1), wherein said dose is administered to said individual at a rate of less than or equal to 75 mg / 2.5 mL / h.

[0165] 16. A method for preventing symptoms of neuronal ceroid lipofuscinosis (CLN2) disease, comprising administering to an individual in need about 10 mL of a composition comprising a dose of about 300 mg of recombinant human tripeptidyl peptidase-1 (rhTPP1), wherein the composition is administered to the individual over a period of about 4 hours.

[0166] 17. A method for preventing symptoms of neuronal ceroid lipofuscinosis (CLN2) disease, comprising administering to an individual in need a dose of about 300 mg of recombinant human tripeptidyl peptidase-1 (rhTPP1), wherein said dose is administered to said individual at a rate of less than or equal to 75 mg / h.

[0167] 18. A method for preventing symptoms of neuronal ceroid lipofuscinosis (CLN2) disease, comprising administering to an individual in need a dose of about 300 mg of recombinant human tripeptidyl peptidase-1 (rhTPP1), wherein said dose is administered to said individual at a rate of less than or equal to 75 mg / 2.5 mL / h.

[0168] 19. A composition comprising recombinant human tripeptidyl peptidase-1 (rhTPP1) for treating an individual with neuronal ceroid lipofuscinosis (CLN2) disease or preventing symptoms of CLN2 disease, said composition comprising:

[0169] i. A composition comprising approximately 10 mL of approximately 300 mg of rhTPP1, wherein said dose is administered to the individual over an period of approximately 4 hours; or

[0170] ii. A dose of approximately 300 mg of rhTPP1, wherein the dose is administered to the individual at a rate of less than or equal to 75 mg / hour; or

[0171] iii. A dose of approximately 300 mg of rhTPP1, wherein the dose is administered to the individual at a rate of less than or equal to 75 mg / 2.5 mL / hour.

[0172] 20. Use of a recombinant human tripeptidyl peptidase-1 (rhTPP1) for manufacturing an agent for treating neuronal ceroid lipofuscinosis (CLN2) disease in an individual or for preventing symptoms of CLN2 disease, wherein the agent comprises:

[0173] i. A composition comprising approximately 10 mL of approximately 300 mg of rhTPP1, wherein said dose is administered to the individual over an period of approximately 4 hours; or

[0174] ii. A dose of approximately 300 mg of rhTPP1, wherein the dose is administered to the individual at a rate of less than or equal to 75 mg / hour; or

[0175] iii. A dose of approximately 300 mg of rhTPP1, wherein the dose is administered to the individual at a rate of less than or equal to 75 mg / 2.5 mL / hour.

[0176] 21. The method, composition or use according to any one of embodiments 10 to 20, wherein the individual has a family history of CLN2 disease.

[0177] 22. The method, composition or use according to any one of embodiments 10 to 21, wherein the rhTPP1 comprises the amino acid sequence of SEQ ID NO:1 or a fragment thereof.

[0178] 23. The method, composition, or use according to any one of embodiments 10 to 22, wherein the formulation, composition, or dose is administered every other week.

[0179] 24. The method, composition, or use according to any one of embodiments 10 to 23, wherein the formulation, composition, or dose is administered intraventricularly.

[0180] 25. The method, composition, or use according to any one of embodiments 10 to 23, wherein the formulation, composition, or dose is administered intrathecally.

[0181] 26. The method, composition, or use according to any one of embodiments 10 to 23, wherein the formulation, composition, or dose is administered intraocularly.

[0182] 27. The method, composition, or use according to any one of embodiments 10 to 23, wherein the formulation, composition, or dose is administered intraventricularly and intraocularly.

[0183] 28. The method, composition, or use according to any one of embodiments 10 to 23, wherein the formulation, composition, or dose is administered intrathecally and intraocularly.

[0184] 29. The method, composition, or use according to any one of embodiments 10 to 28 further includes administering a rinse solution to the individual after administering the formulation, composition, or dose comprising rhTPP1.

[0185] 30. The method, composition, or use according to embodiment 29, comprising administering about 0.5 mL to about 5 mL of the rinse solution to the individual.

[0186] 31. The method, composition, or use according to embodiment 29 or 30, wherein the rinsing solution comprises about 0.11 mg / mL of disodium hydrogen phosphate heptahydrate, about 0.08 mg / mL of sodium dihydrogen phosphate monohydrate, about 8.77 mg / mL of sodium chloride, about 0.22 mg / mL of potassium chloride, about 0.16 mg / mL of magnesium chloride hexahydrate, and about 0.21 mg / mL of calcium chloride dihydrate.

[0187] 32. A method of treating neuronal ceroid lipofuscinosis (CLN2) disease, comprising administering to an individual in need a dose of rhTPP1 that effectively maintains or slows or reduces the decline of physiological function in the individual, wherein the physiological function is language function, motor function, vision or eating function.

[0188] 33. A composition comprising recombinant human tripeptidyl peptidase-1 (rhTPP1) for treating an individual with neuronal ceroid lipofuscinosis (CLN2) disease, said composition comprising a dose of recombinant human tripeptidyl peptidase-1 (rhTPP1) that effectively maintains or slows or reduces the decline of physiological function in said individual, said physiological function being language function, motor function, vision or eating function.

[0189] 34. Use of a recombinant human tripeptidyl peptidase-1 (rhTPP1) for manufacturing a medicament for treating an individual with neuronal ceroid lipofuscinosis (CLN2) disease, wherein the medicament comprises a dose of rhTPP1 that effectively maintains or slows or reduces the decline of physiological function in the individual, wherein the physiological function is language function, motor function, vision or eating function.

[0190] 35. The method, composition or use according to any one of embodiments 32 to 34, wherein the rhTTP1 dose effectively maintains or slows or reduces language function decline in the individual.

[0191] 36. The method, composition or use according to any one of embodiments 32 to 35, wherein the rhTTP1 dose effectively maintains motor function or slows or reduces motor function degeneration in the individual.

[0192] 37. A method for maintaining or slowing or reducing language function decline in an individual suffering from neuronal ceroid lipofuscinosis (CLN2) disease, comprising administering a therapeutically effective dose of recombinant human tripeptidyl peptidase-1 (rhTPP1) to said individual.

[0193] 38. A method for maintaining or slowing or reducing motor function decline in an individual suffering from neuronal ceroid lipofuscinosis (CLN2) disease, comprising administering a therapeutically effective dose of recombinant human tripeptidyl peptidase-1 (rhTPP1) to said individual.

[0194] 39. A composition comprising a therapeutically effective dose of recombinant human tripeptidyl peptidase-1 (rhTPP1), said composition being used for:

[0195] i. To maintain or slow or reduce language function decline in individuals with neuronal ceroid lipofuscinosis (CLN2); or

[0196] ii. To maintain or slow or reduce motor function decline in individuals with neuronal ceroid lipofuscin deposition disease (CLN2).

[0197] 40. Use of a therapeutically effective dose of recombinant human tripeptidyl peptidase-1 (rhTPP1) for the manufacture of a pharmaceutical agent for the following purposes:

[0198] i. To maintain or slow or reduce language function decline in individuals with neuronal ceroid lipofuscinosis (CLN2); or

[0199] ii. To maintain or slow or reduce motor function decline in individuals with neuronal ceroid lipofuscin deposition disease (CLN2).

[0200] 41. The method, composition, or use according to any one of embodiments 32 to 40, wherein the language or motor function deterioration is a reduction of at least one point compared to a previous rating determined before or during treatment, as measured using Weill Cornell Medical College or the Hamburg disease rating scale.

[0201] 42. The method, composition, or use according to any one of embodiments 32 to 41, wherein the rhTTP1 dose effectively maintains vision or slows or reduces vision deterioration in the individual.

[0202] 43. A method for maintaining or slowing or reducing visual impairment in an individual suffering from neuronal ceroid lipofuscinosis (CLN2) disease, comprising administering a therapeutically effective dose of recombinant human tripeptidyl peptidase-1 (rhTPP1) to said individual.

[0203] 44. A composition comprising a therapeutically effective dose of recombinant human tripeptidyl peptidase-1 (rhTPP1) for maintaining vision or slowing or reducing vision deterioration in individuals suffering from neuronal ceroid lipofuscinosis (CLN2).

[0204] 45. Use of a therapeutically effective amount of recombinant human tripeptidyl peptidase-1 (rhTPP1) for the manufacture of an agent for maintaining or slowing or reducing visual deterioration in individuals suffering from neuronal ceroid lipofuscinosis (CLN2).

[0205] 46. ​​The method, composition or use according to any one of embodiments 42 to 45, wherein the visual impairment is a reduction of at least one point compared to baseline, as measured using the Hamburger Disease Rating Scale.

[0206] 47. The method, composition or use according to any one of embodiments 32 to 46, wherein the rhTTP1 dose effectively maintains or slows or reduces the decline of eating function in the individual.

[0207] 48. The method, composition, or use according to embodiment 47, wherein the decline in eating function is measured as a reduction of at least one point from baseline, as measured using the Weill Cornell Medical College Disease Rating Scale.

[0208] 49. A method of treating neuronal ceroid lipofuscinosis (CLN2) disease, comprising administering to an individual in need a dose of recombinant human tripeptidyl peptidase-1 (rhTPP1) that effectively improves physiological function in said individual, wherein said physiological function is language function, motor function, vision or eating function.

[0209] 50. A composition comprising recombinant human tripeptidyl peptidase-1 (rhTPP1) for treating an individual with neuronal ceroid lipofuscinosis (CLN2) disease, said composition comprising a dose of recombinant human tripeptidyl peptidase-1 (rhTPP1) that effectively improves physiological function in said individual, said physiological function being language function, motor function, vision or eating function.

[0210] 51. Use of a recombinant human tripeptidyl peptidase-1 (rhTPP1) for manufacturing a medicament for treating neuronal ceroid lipofuscinosis (CLN2) in an individual, wherein the medicament comprises a dose of rhTPP1 that effectively improves physiological functions in the individual, wherein the physiological functions are language function, motor function, vision or eating function.

[0211] 52. The method, composition, or use according to any one of embodiments 49 to 51, wherein the rhTPP1 dose effectively improves language function in the individual.

[0212] 53. The method, composition, or use according to any one of embodiments 49 to 52, wherein the rhTPP1 dose effectively improves motor function in the individual.

[0213] 54. A method for improving language and / or motor function in an individual with neuronal ceroid lipofuscinosis (CLN2) disease, comprising administering a therapeutically effective dose of recombinant human tripeptidyl peptidase-1 (rhTPP1) to said individual.

[0214] 55. A composition comprising a therapeutically effective dose of recombinant human tripeptidyl peptidase-1 (rhTPP1) for improving language and / or motor function in individuals with neuronal ceroid lipofuscinosis (CLN2) disease.

[0215] 56. Use of a therapeutically effective dose of recombinant human tripeptidyl peptidase-1 (rhTPP1) for the manufacture of a medicament for improving language and / or motor function in individuals with neuronal ceroid lipofuscinosis (CLN2).

[0216] 57. The method, composition, or use according to any one of embodiments 49 to 56, wherein the improvement in language or motor function is an increase of one point or greater as measured using the Weill Cornell Medical College or Hamburger Disease Rating Scale.

[0217] 58. The method, composition, or use according to any one of embodiments 49 to 57, wherein the rhTPP1 dose effectively improves vision.

[0218] 59. A method for improving vision in an individual suffering from neuronal ceroid lipofuscinosis (CLN2) disease, comprising administering a therapeutically effective dose of recombinant human tripeptidyl peptidase-1 (rhTPP1) to said individual.

[0219] 60. A composition comprising a therapeutically effective dose of recombinant human tripeptidyl peptidase-1 (rhTPP1) for improving vision in individuals suffering from neuronal ceroid lipofuscinosis (CLN2) disease.

[0220] 61. Use of a therapeutically effective dose of recombinant human tripeptidyl peptidase-1 (rhTPP1) for the manufacture of a medicament for improving vision in individuals suffering from neuronal ceroid lipofuscinosis (CLN2).

[0221] 62. The method, composition, or use according to any one of embodiments 58 to 61, wherein the improvement in vision is an increase of one point or greater as measured using the Hamburger Disease Rating Scale.

[0222] 63. The method, composition or use according to any one of embodiments 49 to 62, wherein the rhTPP1 dose effectively improves eating function.

[0223] 64. The method, composition, or use according to embodiment 63, wherein the improvement in eating is an increase equal to or greater than 1.0 as measured using the Weill Cornell Medical College Disease Rating Scale.

[0224] 65. A method of treating neuronal ceroid lipofuscinosis (CLN2) disease, comprising administering to an individual in need a dose of recombinant human tripeptidyl peptidase-1 (rhTPP1) that is effective in preventing or improving neurological symptoms of said disease, wherein said neurological symptoms are decreased brain volume, reduced gray matter in the brain, seizures, or increased cerebrospinal fluid.

[0225] 66. A composition comprising recombinant human tripeptidyl peptidase-1 (rhTPP1) for treating an individual with neuronal ceroid lipofuscinosis (CLN2) disease, said composition comprising a dose of rhTPP1 effective in preventing or improving neurological symptoms of said disease, said neurological symptoms being decreased brain volume, reduced gray matter in the brain, seizures, or increased cerebrospinal fluid.

[0226] 67. Use of a recombinant human tripeptidyl peptidase-1 (rhTPP1) for the manufacture of a medicament for treating an individual with neuronal cerebrospinal fluid deposition disease (CLN2), wherein the medicament comprises a dose of rhTPP1 effective in preventing or improving neurological symptoms of the disease, wherein the neurological symptoms are decreased brain volume, decreased gray matter in the brain, seizures, or increased cerebrospinal fluid.

[0227] 68. The method, composition, or use according to any one of embodiments 65 to 67, wherein the rhTTP1 dose effectively maintains or slows or reduces the decline in brain volume in the individual.

[0228] 69. A method for maintaining or slowing or reducing brain volume decline in an individual suffering from neuronal ceroid lipofuscinosis (CLN2) disease, comprising administering a therapeutically effective dose of recombinant human tripeptidyl peptidase-1 (rhTPP1) to said individual.

[0229] 70. A composition comprising a therapeutically effective dose of recombinant human tripeptidyl peptidase-1 (rhTPP1) for maintaining or slowing or reducing brain mass loss in an individual suffering from neuronal ceroid lipofuscinosis (CLN2) disease.

[0230] 71. Use of a therapeutically effective dose of recombinant human tripeptidyl peptidase-1 (rhTPP1) for the manufacture of an agent for maintaining or slowing or reducing the decline in brain volume in an individual suffering from neuronal ceroid lipofuscinosis (CLN2).

[0231] 72. The method, composition, or use according to any one of embodiments 65 to 71, wherein the rhTTP1 dose effectively maintains or slows or reduces gray matter degeneration in the individual's brain.

[0232] 73. The method, composition, or use according to any one of embodiments 65 to 72, wherein the rhTPP1 dose effectively reduces the number of seizures experienced by the individual each month.

[0233] 74. The method, composition, or use according to any one of embodiments 65 to 73, wherein the rhTTP1 dose effectively maintains or slows the increase in cerebrospinal fluid volume in the individual.

[0234] 75. The method, composition or use according to any one of embodiments 32 to 74, wherein the rhTPP1 comprises the amino acid sequence of SEQ ID NO:1 or a fragment thereof.

[0235] 76. The method, composition, or use according to any one of embodiments 32 to 75, wherein the dose is about 10 mL of a composition comprising about 300 mg rhTPP1, administered to the individual over a period of about 4 hours.

[0236] 77. The method, composition or use according to any one of embodiments 32 to 76, wherein the dose is about 300 mg rhTPP1, administered to the individual at a rate of less than or equal to about 75 mg / hr.

[0237] 78. The method, composition or use according to any one of embodiments 32 to 77, wherein the dose is about 300 mg rhTPP1, administered to the individual at a rate of less than or equal to about 75 mg / 2.5 mL / hour.

[0238] 79. The method, composition, or use according to any one of embodiments 32 to 78, wherein the dose is administered every other week.

[0239] 80. The method, composition, or use according to any one of embodiments 32 to 79, wherein the dose is administered intraventricularly.

[0240] 81. The method, composition, or use according to any one of embodiments 32 to 79, wherein the dose is administered intrathecally.

[0241] 82. The method, composition, or use according to embodiment 80 or 81, wherein the dose is administered without removal of the cerebrospinal fluid from the individual before administration of the dose of rhTPP1.

[0242] 83. The method, composition, or use according to any one of embodiments 32 to 79, wherein the dose is administered intraocularly.

[0243] 84. The method, composition or use according to any one of embodiments 32 to 79, wherein the dose is administered intraventricularly and intraocularly.

[0244] 85. The method, composition, or use according to any one of embodiments 10 to 84, wherein the dosage is administered as a formulation comprising potassium chloride at a concentration of about 0.1 mg / mL to about 1 mg / mL, magnesium chloride hexahydrate at a concentration of about 0.1 mg / mL to about 1 mg / mL, and calcium chloride dihydrate at a concentration of about 0.1 mg / mL to about 1 mg / mL.

[0245] 86. The method, composition, or use according to any one of embodiments 10 to 85, wherein the dosage is administered as a formulation comprising a concentration of disodium hydrogen phosphate heptahydrate of about 0.01 mg / mL to about 1 mg / mL, sodium dihydrogen phosphate monohydrate of about 0.01 mg / mL to about 1 mg / mL, and sodium chloride of about 1 mg / mL to about 20 mg / mL.

[0246] 87. The method, composition, or use according to any one of embodiments 10 to 86, wherein the dosage is administered as a formulation comprising about 0.11 mg / mL of disodium hydrogen phosphate heptahydrate, about 0.08 mg / mL of sodium dihydrogen phosphate monohydrate, about 8.77 mg / mL of sodium chloride, about 0.22 mg / mL of potassium chloride, about 0.16 mg / mL of magnesium chloride hexahydrate, and about 0.21 mg / mL of calcium chloride dihydrate.

[0247] 88. The method, composition, or use according to any one of embodiments 32 to 87 further includes administering a flushing solution to the individual after administering the dose comprising rhTPP1.

[0248] 89. The method, composition, or use according to embodiment 88, comprising administering about 0.5 mL to about 5 mL of the rinse solution to the individual.

[0249] 90. The method, composition, or use according to embodiment 88 or 89, wherein the rinsing solution comprises about 0.11 mg / mL of disodium hydrogen phosphate heptahydrate, about 0.08 mg / mL of sodium dihydrogen phosphate monohydrate, about 8.77 mg / mL of sodium chloride, about 0.22 mg / mL of potassium chloride, about 0.16 mg / mL of magnesium chloride hexahydrate, and about 0.21 mg / mL of calcium chloride dihydrate.

[0250] 91. A kit comprising a formulation and a rinsing solution according to any one of embodiments 1 to 9.

[0251] 92. The kit according to embodiment 91, wherein the rinsing solution comprises about 0.11 mg / mL of disodium hydrogen phosphate heptahydrate, about 0.08 mg / mL of sodium dihydrogen phosphate monohydrate, about 8.77 mg / mL of sodium chloride, about 0.22 mg / mL of potassium chloride, about 0.16 mg / mL of magnesium chloride hexahydrate, and about 0.21 mg / mL of calcium chloride dihydrate.

[0252] 93. The kit according to embodiment 91 or 92 further includes a reservoir and catheter for implantation.

[0253] 94. The kit according to any one of embodiments 91 to 93 further comprises one or more elements selected from the group consisting of: extension tubing, in-line filter, port needle, syringe, syringe needle, and combinations thereof.

[0254] All disclosures, patents, and patent applications referenced in this specification are incorporated herein by reference as if specifically and individually indicated that each individual disclosure, patent, or patent application is incorporated by reference. Although the invention has been described in considerable detail by description and examples for clarity, it will be apparent to those skilled in the art that certain variations and modifications may be made without departing from the spirit or scope of the appended claims, based on the teachings of the invention. sequence list <110> BIOMARIN PHARMACEUTICAL INC. <120> TPP1 formulations and methods for treating CLN2 disease <130> 30610 / 49443 / PC <160> 2 <170> PatentIn version 3.5 <210> 1 <211> 544 <212> PRT <213> Homo sapiens <400> 1 Ser Tyr Ser Pro Glu Pro Asp Gln Arg Arg Thr Leu Pro Pro Gly Trp 1 5 10 15 Val Ser Leu Gly Arg Ala Asp Pro Glu Glu Glu Leu Ser Leu Thr Phe 20 25 30 Ala Leu Arg Gln Gln Asn Val Glu Arg Leu Ser Glu Leu Val Gln Ala 35 40 45 Val Ser Asp Pro Ser Ser Pro Gln Tyr Gly Lys Tyr Leu Thr Leu Glu 50 55 60 Asn Val Ala Asp Leu Val Arg Pro Ser Pro Leu Thr Leu His Thr Val 65 70 75 80 Gln Lys Trp Leu Leu Ala Ala Gly Ala Gln Lys Cys His Ser Val Ile 85 90 95 Thr Gln Asp Phe Leu Thr Cys Trp Leu Ser Ile Arg Gln Ala Glu Leu 100 105 110 Leu Leu Pro Gly Ala Glu Phe His His Tyr Val Gly Gly Pro Thr Glu 115 120 125 Thr His Val Val Arg Ser Pro His Pro Tyr Gln Leu Pro Gln Ala Leu 130 135 140 Ala Pro His Val Asp Phe Val Gly Gly Leu His Arg Phe Pro Pro Thr 145 150 155 160 Ser Ser Leu Arg Gln Arg Pro Glu Pro Gln Val Thr Gly Thr Val Gly 165 170 175 Leu His Leu Gly Val Thr Pro Ser Val Ile Arg Lys Arg Tyr Asn Leu 180 185 190 Thr Ser Gln Asp Val Gly Ser Gly Thr Ser Asn Asn Ser Gln Ala Cys 195 200 205 Ala Gln Phe Leu Glu Gln Tyr Phe His Asp Ser Asp Leu Ala Gln Phe 210 215 220 Met Arg Leu Phe Gly Gly Asn Phe Ala His Gln Ala Ser Val Ala Arg 225 230 235 240 Val Val Gly Gln Gln Gly Arg Gly Arg Ala Gly Ile Glu Ala Ser Leu 245 250 255 Asp Val Gln Tyr Leu Met Ser Ala Gly Ala Asn Ile Ser Thr Trp Val 260 265 270 Tyr Ser Ser Pro Gly Arg His Glu Gly Gln Glu Pro Phe Leu Gln Trp 275 280 285 Leu Met Leu Leu Ser Asn Glu Ser Ala Leu Pro His Val His Thr Val 290 295 300 Ser Tyr Gly Asp Asp Glu Asp Ser Leu Ser Ser Ala Tyr Ile Gln Arg 305 310 315 320 Val Asn Thr Glu Leu Met Lys Ala Ala Ala Arg Gly Leu Thr Leu Leu 325 330 335 Phe Ala Ser Gly Asp Ser Gly Ala Gly Cys Trp Ser Val Ser Gly Arg 340 345 350 His Gln Phe Arg Pro Thr Phe Pro Ala Ser Ser Pro Tyr Val Thr Thr 355 360 365 Val Gly Gly Thr Ser Phe Gln Glu Pro Phe Leu Ile Thr Asn Glu Ile 370 375 380 Val Asp Tyr Ile Ser Gly Gly Gly Phe Ser Asn Val Phe Pro Arg Pro 385 390 395 400 Ser Tyr Gln Glu Glu Ala Val Thr Lys Phe Leu Ser Ser Ser Pro His 405 410 415 Leu Pro Pro Ser Ser Tyr Phe Asn Ala Ser Gly Arg Ala Tyr Pro Asp 420 425 430 Val Ala Ala Leu Ser Asp Gly Tyr Trp Val Val Ser Asn Arg Val Pro 435 440 445 Ile Pro Trp Val Ser Gly Thr Ser Ala Ser Thr Pro Val Phe Gly Gly 450 455 460 Ile Leu Ser Leu Ile Asn Glu His Arg Ile Leu Ser Gly Arg Pro Pro 465 470 475 480 Leu Gly Phe Leu Asn Pro Arg Leu Tyr Gln Gln His Gly Ala Gly Leu 485 490 495 Phe Asp Val Thr Arg Gly Cys His Glu Ser Cys Leu Asp Glu Glu Val 500 505 510 Glu Gly Gln Gly Phe Cys Ser Gly Pro Gly Trp Asp Pro Val Thr Gly 515 520 525 Trp Gly Thr Pro Asn Phe Pro Ala Leu Leu Lys Thr Leu Leu Asn Pro 530 535 540 <210> 2 <211> 368 <212> PRT <213> Homo sapiens <400> 2 Leu His Leu Gly Val Thr Pro Ser Val Ile Arg Lys Arg Tyr Asn Leu 1 5 10 15 Thr Ser Gln Asp Val Gly Ser Gly Thr Ser Asn Asn Ser Gln Ala Cys 20 25 30 Ala Gln Phe Leu Glu Gln Tyr Phe His Asp Ser Asp Leu Ala Gln Phe 35 40 45 Met Arg Leu Phe Gly Gly Asn Phe Ala His Gln Ala Ser Val Ala Arg 50 55 60 Val Val Gly Gln Gln Gly Arg Gly Arg Ala Gly Ile Glu Ala Ser Leu 65 70 75 80 Asp Val Gln Tyr Leu Met Ser Ala Gly Ala Asn Ile Ser Thr Trp Val 85 90 95 Tyr Ser Ser Pro Gly Arg His Glu Gly Gln Glu Pro Phe Leu Gln Trp 100 105 110 Leu Met Leu Leu Ser Asn Glu Ser Ala Leu Pro His Val His Thr Val 115 120 125 Ser Tyr Gly Asp Asp Glu Asp Ser Leu Ser Ser Ala Tyr Ile Gln Arg 130 135 140 Val Asn Thr Glu Leu Met Lys Ala Ala Ala Arg Gly Leu Thr Leu Leu 145 150 155 160 Phe Ala Ser Gly Asp Ser Gly Ala Gly Cys Trp Ser Val Ser Gly Arg 165 170 175 His Gln Phe Arg Pro Thr Phe Pro Ala Ser Ser Pro Tyr Val Thr Thr 180 185 190 Val Gly Gly Thr Ser Phe Gln Glu Pro Phe Leu Ile Thr Asn Glu Ile 195 200 205 Val Asp Tyr Ile Ser Gly Gly Gly Phe Ser Asn Val Phe Pro Arg Pro 210 215 220 Ser Tyr Gln Glu Glu Ala Val Thr Lys Phe Leu Ser Ser Ser Pro His 225 230 235 240 Leu Pro Pro Ser Ser Tyr Phe Asn Ala Ser Gly Arg Ala Tyr Pro Asp 245 250 255 Val Ala Ala Leu Ser Asp Gly Tyr Trp Val Val Ser Asn Arg Val Pro 260 265 270 Ile Pro Trp Val Ser Gly Thr Ser Ala Ser Thr Pro Val Phe Gly Gly 275 280 285 Ile Leu Ser Leu Ile Asn Glu His Arg Ile Leu Ser Gly Arg Pro Pro 290 295 300 Leu Gly Phe Leu Asn Pro Arg Leu Tyr Gln Gln His Gly Ala Gly Leu 305 310 315 320 Phe Asp Val Thr Arg Gly Cys His Glu Ser Cys Leu Asp Glu Glu Val 325 330 335 Glu Gly Gln Gly Phe Cys Ser Gly Pro Gly Trp Asp Pro Val Thr Gly 340 345 350 Trp Gly Thr Pro Asn Phe Pro Ala Leu Leu Lys Thr Leu Leu Asn Pro 355 360 365

Claims

1. A kit comprising a 300 mg dose of rhTPP1 of the sequence shown in SEQ ID NO: 1 or SEQ ID NO: 2, said rhTPP1 being contained in a composition for administration, said composition for administration comprising a formulation of disodium hydrogen phosphate heptahydrate at a concentration of 0.11 mg / mL, sodium dihydrogen phosphate monohydrate at a concentration of 0.08 mg / mL, sodium chloride at a concentration of 8.77 mg / mL, potassium chloride at a concentration of 0.22 mg / mL, magnesium chloride hexahydrate at a concentration of 0.16 mg / mL, and calcium chloride dihydrate at a concentration of 0.21 mg / mL, said kit further comprising a rinse solution at pH 6.

5.

2. The kit according to claim 1, wherein the rinsing solution comprises 0.11 mg / mL of disodium hydrogen phosphate heptahydrate, 0.08 mg / mL of sodium dihydrogen phosphate monohydrate, 8.77 mg / mL of sodium chloride, 0.22 mg / mL of potassium chloride, 0.16 mg / mL of magnesium chloride hexahydrate, and 0.21 mg / mL of calcium chloride dihydrate.

3. The kit according to claim 1 or 2, further comprising a reservoir and catheter for implantation.

4. The kit according to claim 1 or 2, further comprising one or more elements selected from the group consisting of: extension tubing, in-line filter, port needle, syringe, syringe needle, and combinations thereof.

5. The kit of claim 3, further comprising one or more elements selected from the group consisting of: extension tubing, in-line filter, port needle, syringe, syringe needle, and combinations thereof.

Images