Composition for the treatment of sanfilippo syndrome type a (mps iiia) comprising heparan n-sulfatase (HNS)

Abstract

The present invention relates to a pharmaceutical composition comprising heparan N-sulfatase (HNS) for preventing or treating Swefil syndrome type A (MPS IIIA), and more particularly, to an optimal dose and cycle for administering heparan N-sulfatase which can effectively reduce the accumulation of heparan sulfate (HS) while improving the cognition of a patient, and more particularly, to a pharmaceutical composition for preventing or treating Swefil syndrome type A (MPS IIIA) comprising heparan N-sulfatase (HNS) for preventing or treating Swefil syndrome type A (MPS IIIA). According to the invention, it can be used as an enzyme replacement therapy (ERT) for the treatment of Swefil Syndrome Type A.

Description

(19) State Intellectual Property Office (12) Invention Patent Application (10) Application Publication Number (43) Application Publication Date (21) Application Number 202480054559.4 (22) Application Date 2024.08.28 (30) Priority Data 10-2023-0112883 2023.08.28 KR (85) PCT International Application Entering National Phase Date 2026.02.25 (86) PCT International Application Application Data PCT / KR2024 / 012852 2024.08.28 (87) PCT International Application Publication Data WO2025 / 048486 EN 2025.03.06 (71) Applicant: Green Cross Co., Ltd. Address: Gyeonggi Province, South Korea (72) Inventors: Kim So-ra, Kim Min-joo, Chen Dong-kui (74) Patent Agency: Beijing Kunrui Law Firm 11494 Patent Attorney Chen An (51) Int.Cl. A61K 38 / 46 (2006.01) A61K 9 / 00 (2006.01) A61K 9 / 08 (2006.01) A61K 47 / 26 (2006.01) A61K 47 / 30 (2006.01) A61P 25 / 28 (2006.01) (54) Invention Title Composition for the Treatment of San Phillips Syndrome Type A (MPS IIIA) Including Heparin N-sulfatase (HNS) (57) Abstract This invention relates to a pharmaceutical composition comprising heparin N-sulfatase (HNS) for the prevention or treatment of San Phillips Syndrome Type A (MPS IIIA), and more specifically, to the optimal dose and cycle of administration of heparin N-sulfatase, which effectively reduces the accumulation of heparin sulfate (HS) while improving patient cognition. According to the present invention, it can be used as an enzyme replacement therapy (ERT) for the treatment of Santa Felipe syndrome type A. Claims 1 page, Description 16 pages, Sequence List (electronic publication), Drawings 10 pages. CN 122121891 A 2026.05.29 CN 1 22 12 18 91 A 1. A pharmaceutical composition for the prevention or treatment of Santa Felipe syndrome type A (MPS IIIA), comprising heparin N-sulfatase (HNS), wherein the heparin N-sulfatase is administered to the patient at a dose of 3 to 150 mg per dose, at intervals of 2 to 4 weeks, via intraventricular injection (ICV), intracerebral injection (IC), or intrathecal injection (IT). 2. The pharmaceutical composition according to claim 1, wherein the heparin N-sulfatase is administered at a dose of 6 to 100 mg per dose. 3. The pharmaceutical composition according to claim 1, wherein the total volume of each dose is 10 mL or less.4. The pharmaceutical composition of claim 1, wherein the pharmaceutical composition further comprises 1 to 40 mM histidine buffer. 5. The pharmaceutical composition of claim 1, wherein the pharmaceutical composition further comprises a sugar. 6. The pharmaceutical composition of claim 5, wherein the sugar is at least one selected from the group consisting of trehalose, sucrose, maltose, lactose, and sorbitol. 7. The pharmaceutical composition of claim 6, wherein the concentration of the sugar is 0.1 to 5.0 w / v%. 8. The pharmaceutical composition of claim 1, wherein the composition further comprises a salt. 9. The pharmaceutical composition of claim 8, wherein the salt is NaCl or KCl. 10. The pharmaceutical composition of claim 9, wherein the concentration of the salt is 30 mM to 500 mM. 11. The pharmaceutical composition of claim 1, wherein the composition further comprises a surfactant. 12. The pharmaceutical composition of claim 11, wherein the surfactant is polysorbate 20 or polysorbate 80. 13. The pharmaceutical composition of claim 12, wherein the concentration of the surfactant is 0.0001 to 0.1 w / v%. 14. The pharmaceutical composition of claim 1, wherein the drug is administered to the central nervous system via intraventricular injection (ICV). Claims 1 / 1 page 2 CN 122121891 A Composition comprising heparin N-sulfatase (HNS) for the treatment of Santa Felipe syndrome type A (MPS IIIA) Technical Field

[0001] The present invention relates to a pharmaceutical composition comprising heparin N-sulfatase (HNS) for the prevention or treatment of Santa Felipe syndrome type A (MPS IIIA), and more particularly, to a pharmaceutical composition characterized by administration of HNS at specific doses and cycles, and a method of using the pharmaceutical composition for the prevention or treatment of MPS IIIA. Background Art

[0002] Lysosomal storage disease (LSD) is a hereditary metabolic disorder caused by defects in lysosomal function. LSD is caused by lysosomal dysfunction due to a deficiency of one or more enzymes required for the metabolism of lipids, glycoproteins, or mucopolysaccharides. The deficiency of lysosomal enzymes leads to the accumulation of lipids, glycoproteins, or mucopolysaccharides in lysosomes, resulting in systemic abnormalities (Nature Reviews Disease Primers. 4 (1): 27; Biochem. Soc. Trans. 28 (2): 150–4). Mucopolysaccharide storage disease (MPS), or mucopolysaccharide deposition, is a type of LSD caused by the accumulation of glycosaminoglycans (GAGs) in lysosomes due to a deficiency of lysosomal enzymes required for GAG degradation.

[0003] St. Philipp syndrome is a type of MPS named after Dr. St. Philipp, an American physician who first discovered the condition in 1963. Also known as MPS III, St. Philipp syndrome is an autosomal recessive genetic disorder clinically characterized by the absence of corneal opacity, mild physical changes such as hepatomegaly and skeletal changes, but with very severe and progressive central nervous system (CNS) symptoms.

[0004] St. Philipp syndrome is caused by a deficiency of four different enzymes required to break down polysaccharides (particularly GAG). Based on the deficient enzymes, St. Philipp syndrome is classified into MPS IIIA (S. Philipp A), MPS IIIB (S. Philipp B), MPS IIIC (S. Philipp C), and MPS IIID (S. Philipp D). The deficient enzymes and genetic loci for each type of St. Philipp syndrome are listed below.

[0005] Type A (MPS IIIA): Heparin N-sulfatase - Chromosome 17 (17q25.3)

[0006] Type B (MPS IIIB): N-acetyl-α-D-glucosidase - Chromosome 17 (17q21)

[0007] Type C (MPS IIIC): Acetyl-CoA:α-glucosinolate-N-acetyltransferase - Chromosome 14

[0008] Type D (MPS IIID): N-acetyl-α-D-glucosinolate-6-sulfatase - Chromosome 12 (12q14)

[0009] MPS IIIA is caused by a deficiency of heparin N-sulfatase (HNS), an enzyme involved in the breakdown of heparin sulfate (HS) by hydrolyzing the sulfate ester moiety attached to the amino group of the glucosamine residue of HS. Symptoms of MPS IIIA typically appear between the ages of 2 and 6, but some cases are diagnosed after the age of 13. In general, MPS IIIA patients have significant developmental delays and are known to have low long-term survival rates.

[0010] Currently, there are no approved treatments for MPS IIIA, and only palliative care is used to alleviate symptoms. Enzyme replacement therapy (ERT), which involves administering exogenously manufactured HNS to MPS IIIA patients, holds promise for being extremely useful in the treatment of this disease.

[0011] ERT, which involves administering a lacking functional lysosomal enzyme to correct the deficiency of enzyme function, is one of the main therapies for LSD, and its advantage lies in relieving symptoms and preventing permanent damage to the body through simple injection. As a well-known ERT for enzyme storage diseases, intravenous (IV) therapy using glucocerebrosidase (GCase) was first approved by the FDA for Gaucher disease in 1991 (National Gaucher Foundation. Retrieved 2017-06-08).Instructions for Use, Page 1 / 16, CN 122121891 A

[0012] However, given that most LSDs are in the CNS, particularly in neurons and meninges of the brain, causing excessive accumulation of GAGs and leading to various CNS disorders, intravenously administered ERTs are not effective in treating neurological disorders and diseases caused by lysosomal accumulation, especially in the brain, because lysosomal enzymes, as active ingredients, have difficulty crossing the blood-brain barrier (BBB), resulting in insufficient delivery of enzymes to the CNS. Therefore, various CNS delivery therapies that deliver drugs directly to the CNS to bypass the BBB to deliver enzymes are being investigated.

[0013] Several therapies have been developed to deliver drugs to the CNS by bypassing the BBB. In particular, intracerebral injection (IC), intraventricular injection (ICV), and intrathecal injection (IT) are the most common routes of administration for the direct delivery of proteins to the brain.

[0014] Injection of heparin N-sulfatase (HNS) into the CNS has become a method for delivering the enzyme to MPS and has shown significant reductions in GAG and significant improvements in neurological symptoms in various animal models of MPS (Molecular Therapy - Methods & Clinical Development, 21, 67-75). However, due to the very limited dosage of direct brain injection therapy, there is a need to develop ERTs with appropriate effective doses and cycles to achieve effective levels of therapeutic benefit.

[0015] Against this technical background, the inventors of the present invention have determined the optimal dose and cycle for administering HNS to the CNS, which has significant effects, and have completed the present invention.

[0016] The information provided in the Background section is intended only to enhance the understanding of the background of the invention and may not include information constituting prior art known to those skilled in the art. Summary of the Invention

[0017] The object of the present invention is to provide a method for preventing or treating San Phillips syndrome type A (MPS IIIA) by administering heparin N-sulfatase (HNS) at a sustained and stable effective dose and cycle.

[0018] To achieve the above objectives, the present invention provides a pharmaceutical composition comprising heparin N-sulfatase (HNS) for the prevention or treatment of Santa Felipe syndrome type A (MPS IIIA), wherein the heparin N-sulfatase is administered to a patient at a dose of 3 to 150 mg per dose, at intervals of 2 to 4 weeks, via intraventricular injection (ICV), intracerebral injection (IC), or intrathecal injection (IT).

[0019] The present invention also provides a method for the prevention or treatment of Santa Felipe syndrome type A (MPS IIIA), the method comprising administering the pharmaceutical composition to a patient.

[0020] The present invention also provides the use of the pharmaceutical composition for the prevention or treatment of Santa Felipe syndrome type A (MPS IIIA).

[0021] The present invention also provides the use of the pharmaceutical composition in the preparation of a medicament for the prevention or treatment of Santa Felipe syndrome type A (MPS IIIA). Brief Description of the Drawings

[0022] Figure 1 is a graph showing the results of a single-dose efficacy test of GC1130A, wherein Figure 1A shows the HS content in the brain (all data are expressed as mean ± SEM. **P < 0.01, ***P < 0.0005, ***P < 0.0001 compared with vector-treated MPS IIIA mice), and Figure 1B shows the HS content in cerebrospinal fluid (CSF) (all data are expressed as mean ± SEM. **P < 0.01, ***P < 0.0005, ***P < 0.0001 compared with vector-treated MPS IIIA mice).

[0023] Figure 2 is a graph showing the results of a low-dose, single-dose efficacy test of GC1130A, where Figure 2A shows the HS content in the brain (all data are expressed as mean ± SEM, *** P < 0.001, **** P < 0.0001 compared to G2), and Figure 2B shows the HS content in the CSF (all data are expressed as mean ± SEM. * P < 0.05, ** P < 0.002, **** P < 0.0001 compared to G2).

[0024] Figure 3 is a graph showing the correlation between HS in the brain and CSF when administered as a single dose via ICV.

[0025] Figure 4 is a graph showing the changes in HS content in the brain and CSF with GC1130A dose after repeated administration (analyzed using GraphPad Prism version 9.4.0, by one-way ANOVA Dunnett's multiple comparison test, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 VS vector group, (mean ± SEM)).

[0026] Figure 5 is a graph showing the correlation of HS in the brain and CSF after repeated ICV administration.

[0027] Figure 6 is a graph showing the results of an open field test assessing the behavior of mice after repeated administration of GC1130A.

[0028] Figure 7 shows the biodistribution of GC1130A conjugated with fluorescent dye in mice after IV or ICV administration (the graph in Figure 7B is for each time curve n=4).

[0029] Figure 8 is the time curve of ICV administration of GC1130A in the mouse brain, where the error bars represent the standard deviation of the mean (n=4).

[0030] Figure 9 is a graph showing the quantitative results of repeated ICV administration of GC1130A to the mouse brain.

[0031] Figure 10 is a graph showing the results of a quantitative IHC analysis, namely a LAMP2 analysis, of brain pathological changes after repeated ICV administration.

[0032] Figure 11 is a graph showing the results of a quantitative IHC analysis, namely a CD68 analysis, of brain pathological changes after repeated ICV administration. Detailed Description

[0033] Unless otherwise defined, all technical and scientific terms used herein will have the same meaning as commonly understood by those skilled in the art. Generally, the terms used herein are those well known and commonly used in the art.

[0034] In embodiments of the invention, in order to determine the effective dose and interval of HNS as an ERT, it was observed that in a Santa Felipe syndrome type A (MPS IIIA) mouse model, administration of a single dose of 15 to 60 μg at intervals of 2 to 4 weeks resulted in a significant reduction in HS levels and an improvement in behavioral indicators.

[0035] Furthermore, by comparing allometric growth scale values ​​based on brain weight and CSF, it was determined that a single dose of 3 to 150 mg, preferably 6 to 100 mg, and even more preferably 6 to 60 mg, for human patients when administered at 2 to 4-week intervals was determined.

[0036] Therefore, in one aspect, the present invention relates to a pharmaceutical composition comprising heparin N-sulfatase (HNS) for the prevention or treatment of San Phillips syndrome type A (MPS IIIA), wherein the heparin N-sulfatase is administered to the patient at a dose of 3 to 150 mg per dose at 2 to 4-week intervals via intraventricular injection (ICV), intracerebral injection (IC), or intrathecal injection (IT).

[0037] HNS is a lysosomal enzyme that catalyzes the hydrolysis of HS and N-linked sulfate groups from the non-reducing terminal glucosamine moiety of HS or heparin (Biochem. Biophys. Res. Commun. 2001, 280, 1251-1257). It is well known that mutations in the heparin N-sulfatase gene (SGSH) cause MPS type A (MPSIIIA, OMIM# 252900), also known as San Phillips A syndrome. MPS type A is characterized by the deficiency of heparin N-sulfatase (HNS), an enzyme involved in the lysosomal catabolism of glycosaminoglycan (GAG) heparin sulfate (Neufeld EF, et al. The Metabolic and Molecular Bases of Inherited Disease (2001) pp. 3421-3452). In the absence of this enzyme, GAG accumulates in the lysosomes of neurons and glial cells, causing severe neurological damage.

[0038] As used herein, the term "heparin N-sulfatase" may be used interchangeably with N-sulfoglucosylhydrogenase (SGSH).Specification 3 / 16 pages 5 CN 122121891 A

[0039] In this invention, heparin N-sulfatase may have a wild-type or naturally occurring amino acid sequence. For example, heparin N-sulfatase may be derived from various organisms, more preferably from humans, but is not limited thereto. In this invention, heparin N-sulfatase may include the amino acid sequence represented by SEQ ID NO: 1, but is not limited thereto. In this invention, heparin N-sulfatase may include a sequence having at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% homology with a wild-type or naturally occurring sequence.

[0040]

[0041] In this invention, heparin N-sulfatase may be a recombinant enzyme produced by recombinant synthesis. Techniques known in the art for producing recombinant cells for expressing various target proteins can facilitate the recombinant synthesis of heparin N-sulfatase.

[0042] In this invention, heparin N-sulfatase may also be included in the form of a fusion protein or conjugate. In this invention, heparin N-sulfatase can be fused or conjugated with portions of receptors and / or lysosomal targeting agents capable of binding to brain cell surfaces to promote cellular uptake or lysosomal targeting. Korean Patent No. 10-2007044 discloses modifications of alternative enzymes such as heparin N-sulfatase.

[0043] In this invention, heparin N-sulfatase is administered to the patient at doses of 3 to 150 mg, preferably 5 to 120 mg, more preferably 6 to 100 mg, and most preferably 6 to 60 mg, at intervals of 2 to 4 weeks, but is not limited thereto.

[0044] In particular, heparin N-sulfatase can also be administered as a single dose of 6 to 60 mg at intervals of 2 to 4 weeks, most preferably at intervals of 2 weeks, but is not limited thereto.

[0045] In this invention, the total volume of the pharmaceutical composition per dose can be 10 mL or less, preferably 9 mL or less, more preferably 6 mL or less, but is not limited thereto.

[0046] In this invention, the pharmaceutical composition may include heparin N-sulfatase at concentrations of about 2 to about 50 mg / mL, preferably about 3 to about 40 mg / mL, more preferably about 5 to about 30 mg / mL, more preferably about 8 to about 25 mg / mL, more preferably about 10 to about 20 mg / mL, and most preferably about 12 to about 15 mg / mL, but is not limited thereto.

[0047] The pharmaceutical composition may further include 1 to 40 mM histidine buffer.

[0048] Although formulations for central nervous system delivery of heparan N-sulfatase comprising phosphate have been reported (e.g., Korean Patent No. 10-2007044), studies have consistently reported negative effects of phosphate-buffered saline on the activity of heparan N-sulfatase (J. Inherit Metab. Dis. 1993;16(2):465-72; and Acta Crystallogr D Biol. Crystallogr. 2014 May;70(Pt 5):1321-35). Therefore, histidine buffer can be used as a stabilizer instead of phosphate for central nervous system delivery of heparan N-sulfatase. Histidine buffer offers several advantages over conventional phosphate buffer, including significantly increased stability due to reduced protein-protein and protein-buffered saline interactions, and a significant reduction in turbidity.

[0049] In the pharmaceutical composition according to the invention, the histidine buffer may be, but is not limited to, about 1 to about 40 mM, preferably about 2 to about 30 mM, more preferably about 3 to about 25 mM, and most preferably about 5 to about 20 mM. The concentration of the above-mentioned histidine buffer is a concentration calculated based on the histidine concentration.

[0050] The pH of the pharmaceutical composition according to the invention may be, but is not limited to, about 7.8 or higher, preferably about 7.8 to about 9.0, more preferably about 7.9 to about 8.9, and most preferably about 8.0 to about 8.8.

[0051] In the invention, the pharmaceutical composition may further include sugars.

[0052] The inclusion of sugars, particularly trehalose, in the pharmaceutical composition results in very high purity (%) and titers, not only when the composition is used as is in liquid form, but also when it is formulated and reconstituted into lyophilized formulations.

[0053] In the invention, the sugar may be at least one selected from the group consisting of trehalose, sucrose, maltose, lactose, and sorbitol. In this invention, the concentration of the sugar included may be about 0.1% or more, about 0.5% or more, about 1.0% or more, or about 1.3% or more, more specifically, about 0.1% to about 5.0%, preferably about 0.5% to about 4.0%, and most preferably about 1.0% to about 3.0%.

[0054] In this invention, unless otherwise stated, the % concentration of each substance refers to w / v%.

[0055] In this invention, the pharmaceutical composition may further include a salt.

[0056] In this invention, the salt may be NaCl or KCl.In this invention, the concentration of the included salt can be from about 30 mM to about 500 mM, preferably from about 50 mM to about 400 mM, more preferably from about 60 mM to about 200 mM, and most preferably from about 80 mM to 150 mM, but is not limited thereto.

[0057] In this invention, the concentration of the included salt can have an appropriate osmotic effect for the central nervous system delivery of the pharmaceutical composition of the present invention. Suitable osmotic concentrations for pharmaceutical formulations for central nervous system delivery are known in the art.

[0058] In this invention, the osmotic concentration of the pharmaceutical composition can be, for example, about 400 mOsmol / kg or less, preferably about 350 mOsmol / kg or less, more preferably about 330 mOsmol / kg or less, more preferably about 300 mOsmol / kg or less, and most preferably about 290 mOsmol / kg or less, but is not limited thereto. In this invention, the permeation concentration of the pharmaceutical formulation can be, for example, about 200 to about 400 mOsmol / kg, preferably about 220 to about 360 mOsmol / kg, more preferably about 250 to about 330 mOsmol / kg, and most preferably about 280 to about 300 mOsmol / kg, but is not limited thereto.

[0059] In this invention, the pharmaceutical composition may further include a surfactant.

[0060] In this invention, the surfactant may be a polysorbate-based surfactant, more preferably polysorbate 20 or polysorbate 80, and most preferably polysorbate 20.

[0061] In this invention, the concentration of the included surfactant can be about 0.0001% to about 0.1%, preferably about 0.002% to about 0.07%, more preferably about 0.003% to about 0.05%, and most preferably about 0.004% to about 0.01%, but is not limited thereto.

[0062] However, when the pharmaceutical composition according to the invention is formulated into a lyophilized dosage form, reconstituted, and administered to a patient, the surfactant may be used in the form included in the solution used for reconstitution, rather than included in the pharmaceutical composition and dosage form used for lyophilization.

[0063] The pharmaceutical composition according to the invention may further include suitable carriers, excipients, and diluents conventionally used in pharmaceutical compositions.

[0064] In particular, pharmaceutical excipients for liquid protein formulations are well known to those skilled in the art.Non-limiting examples include, for example, body solvents or co-solvents; sugars or sugar alcohols such as mannitol, sucrose, sorbitol, fructose, maltose, lactose, or dextran; buffers; preservatives such as benzalkonium chloride, benzyl chloride, tertiary ammonium salts, and chlorhexidine diacetate; carriers such as polyethylene glycol (PEG); antioxidants such as ascorbic acid, sodium metabisulfite, and methionine; chelating agents such as EDTA or citric acid; or biodegradable polymers such as water-soluble polyesters; cryoprotectants; lyophilization protectants; fillers; and stabilizers, and the protein formulations described herein may include other pharmaceutically acceptable carriers, excipients, or stabilizers, such as those described in Remington: "The Science and Practice of Pharmacy" 20th edition, Alfonso R Gennaro, Ed., Lippincott Williams & Wilkins (2000), provided that they do not adversely affect the desired properties of the formulation.

[0065] In this invention, the pharmaceutical composition can be formulated into a pharmaceutical dosage form, such as a liquid dosage form or a lyophilized dosage form.

[0066] Liquid formulations are preferably, but not exclusively, in the form of ampoules or pre-filled syringes.

[0067] Preferably, the pharmaceutical composition can be formulated into a lyophilized formulation. Lyophilized formulations have advantages in terms of storage and transportation, and can be prepared by various freeze-drying methods known in the art, in addition to those methods described in the embodiments of this invention.

[0068] If the composition according to the invention is formulated into a lyophilized form, i.e., a dry powder, it can be reconstituted into a liquid composition for administration. Non-limiting examples of solutions for reconstitution may include ordinary aqueous solutions, saline solutions, etc., and if the composition according to the invention does not contain a surfactant or contains an insufficient amount of surfactant, the solution for reconstitution may contain surfactants such as PS20 or PS80.

[0069] The compositions according to the invention can be administered to the central nervous system via various methods of administration. They can be administered to the central nervous system via intraventricular injection (ICV), intracerebral injection (IC), or intrathecal injection (IT), most preferably via intraventricular injection (ICV).

[0070] As used herein, intraventricular injection refers to the administration of medication by injecting the drug into the ventricles of the brain, which are interconnected hollow spaces within the brain. The advantage of intraventricular injection compared to intracerebral injection is the ability to deliver a larger volume of drug over a larger area. Various techniques for intraventricular injection are known in the art, such as, but not limited to, the Ommaya reservoir developed by Ayub Ommaya as a conventional intraventricular injection device, and various techniques for intraventricular injection are continuously being developed and reported.Various other intraventricular injection devices and techniques known in the art or developed thereafter can be used without limitation for intraventricular injection of the pharmaceutical compositions of the present invention.

[0071] As used herein, intraventricular injection refers to the injection of a drug into the brain tissue itself. Various intraventricular injection techniques are known in the prior art, such as Mathon et al. 2015, which describes intraventricular injection methods in detail.

[0072] As used herein, intrathecal injection refers to injection into the spinal canal. Various intrathecal injection techniques are known in the art and are described in detail, for example, in Lazorthes et al. Advances in Drug Delivery Systems and Applications in Neurosurgery, 143-192 and Omaya et al. Cancer Drug Delivery, 1: 169-179, which serve as representative methods.

[0073] In the present invention, when the pharmaceutical composition is administered via intraventricular injection, a certain amount of cerebrospinal fluid (CSF) can be drained from the ventricles of the subject before administration. Drainage of CSF can prevent an increase in intracranial pressure due to changes in CSF volume after ICV administration.

[0074] Preferably, the total administration volume of the pharmaceutical composition according to the invention via intraventricular (ICV) administration may be 10 mL or less, preferably 9 mL or less, more preferably 6 mL or less, but is not limited thereto. Specification 6 / 16 pages 8 CN 122121891 A

[0075] In this invention, administration of the pharmaceutical composition to the central nervous system can deliver heparan N-sulfatase to various target tissues, such as the brain, spinal cord, and peripheral tissues. In this invention, target tissues include any tissue affected by the lysosomal storage disease to be treated; for example, target tissues may be brain target tissues, spinal cord target tissues, and / or peripheral target tissues, and administration to the central nervous system can provide systemic delivery of heparan N-sulfatase.

[0076] In this invention, administration of the pharmaceutical composition to the central nervous system can achieve therapeutic or clinically effective levels or activities in the various target tissues described herein. As used herein, therapeutic or clinically effective levels or activities refer to levels or activities sufficient to impart a therapeutic effect in the target tissue. For example, a therapeutic or clinically effective level or activity can be an enzyme level or activity sufficient to improve disease-related symptoms (e.g., GAG accumulation) in the target tissue.

[0077] In this invention, administration of the formulation or pharmaceutical composition to the central nervous system can achieve an enzyme level or activity of at least 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 95% of the normal level or activity of heparan N-sulfatase in the target tissue.In this invention, administration of the pharmaceutical composition to the central nervous system can achieve an increase in enzyme levels or activity of at least 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 times compared to a control group (e.g., untreated endogenous levels or activity).

[0078] In this invention, administration of the pharmaceutical composition to the central nervous system can result in a reduction in GAG (e.g., heparan sulfate) storage in brain target tissues, spinal cord neurons, and / or peripheral target tissues. In this invention, GAG storage can be reduced by at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 1, 1.5, or 2 times compared to a negative control group (e.g., GAG storage in subjects before treatment or after administration of the carrier only). In this invention, administration of the pharmaceutical composition to the central nervous system can result in a reduction in vacuolation in neurons. For example, compared with the negative control group, it can cause a reduction of at least 20%, 40%, 50%, 60%, 80%, 90%, 1, 1.5, or 2 times.

[0079] The pharmaceutical composition according to the invention can be administered as a single therapeutic agent or in combination with other therapeutic agents, can be administered sequentially or simultaneously with conventional therapeutic agents, and can be administered in single or multiple doses.

[0080] Furthermore, for patient comfort, the pharmaceutical composition according to the invention is preferably administered at a preferably rapid rate of administration. As an example, the rate of administration of the pharmaceutical composition according to the invention can be, but is not limited to, about 0.1 ml / min or higher, or about 0.5 ml / min or higher, preferably about 1 ml / min or higher, more preferably about 2 ml / min or higher, and most preferably about 5 ml / min or higher.

[0081] As used herein, the term “prevention” means any action by administering the composition to prevent the onset of disease or to delay the progression of disease. Furthermore, as used herein, the term “treatment” means any action by administering the composition to improve or alleviate or cure disease symptoms.

[0082] As used herein, “patient” means a mammal, preferably a human, that suffers from or is at risk of suffering from a condition or disease that can be relieved, suppressed, or cured by administration of the composition according to the invention.

[0083] In another aspect, the present invention relates to a method for the prevention or treatment of Santa Felipe syndrome type A (MPS IIIA), the method comprising administering the pharmaceutical composition to a patient.

[0084] In another aspect, the present invention relates to the use of the pharmaceutical composition for the prevention or treatment of Santa Felipe syndrome type A (MPS IIIA).

[0085] In another aspect, the present invention relates to the use of the pharmaceutical composition in the preparation of a medicament for the prevention or treatment of Santa Felipe syndrome type A (MPS IIIA).

[0086] The invention will now be described in more detail with reference to the following examples.These embodiments are merely illustrative of the invention, and it will be apparent to those skilled in the art that the scope of the invention should not be construed as limited by these embodiments. Specification 7 / 16 pages 9 CN 122121891 A

[0087] The recombinant heparin N-sulfatase (rHNS) used in the embodiments of the present invention is named “GC1130A” or “GC1130A protein”.

[0088] Example 1: High-dose single-dose efficacy study of GC1130A

[0089] The purpose of this study was to evaluate the efficacy of GC1130A as a single intraventricular (ICV) injection for the treatment of San Philippo syndrome type 2 (MPS IIIA) in an MPS IIIA mouse model (C57BL / 6 Sgshmps3a gene WT, homozygous mutant, 2 weeks, male & female, n=12) and to establish dose ranges and intervals for repeated ICV dosing efficacy studies.

[0090] As shown in Table 1 below, the animals were divided into five different groups to evaluate the efficacy of the targeted drug.

[0091] [Table 1]

[0092]

[0093]

[0094] Mice in the treatment and control groups received 12 to 110 μg / dose of GC1130A (22 mg / mL, GC Biopharma) and the carrier, respectively, and there were no deaths, systemic symptoms, or changes in body weight and organ weight in either group after ICV administration. Animals were divided into three dosage groups and necropsy was performed on days 7, 14, and 28 after administration. Brain and cerebrospinal fluid (CSF) were collected and stored in a cryogenic freezer until analysis. The changes in heparan sulfate (HS) content in the brain and CSF over time were analyzed by LC-MS / MS to determine the trend of HS clearance after each administration (Figure 1).

[0095] It can be seen that HS had accumulated in the diseased mice by day 7. At the lowest dose of 12 μg, HS decreased on day 7 and increased on day 14, while at the intermediate doses of 37 μg and 110 μg, HS continued to decrease until day 14 and rebounded on day 28. In other words, a dose-dependent effect was found in both the brain and CSF only on day 14 after administration, and repeated doses of 15, 30, and 60 μg were selected based on the above results.

[0096] After 14 days of treatment, HS showed a significant decreasing trend, and after 28 days, except for the low-dose group, the therapeutic effect of the drug weakened but remained unchanged, so further experiments were conducted at dosing intervals of every two weeks (2QW) or every four weeks (4QW).

[0097] Example 2: Study on the efficacy of a low-dose single-dose administration of GC1130A

[0098] The purpose of this study was to evaluate the efficacy of a single low-dose intraventricular (ICV) injection of GC1130A for the treatment of Santa Felipe syndrome type A (MPS IIIA) in a mouse model of MPS IIIA (C57BL / 6 Sgshmps3a gene WT, homozygous mutant, 2 weeks old, male & female, n=14).

[0099] As shown in Table 2 below, the animals were divided into five different groups to evaluate the efficacy of the targeted drug.

[0100] [Table 2]

[0101]

[0102] Mice in the treatment and control groups received 1.5 to 15 μg / dose of GC1130A (15 mg / mL, GC Biopharma) and the carrier, respectively, and there were no deaths, systemic symptoms, or changes in body weight and organ weight in either group after ICV administration. Autopsies were performed on the treatment group at 14 and 28 days post-administration, and brain and cerebrospinal fluid (CSF) were collected and stored in a cryogenic freezer until analysis. Changes in heparan sulfate (HS) content in the brain and CSF over time were analyzed by LC-MS / MS to determine the trend of HS clearance after specific dose administration (Figure 2).

[0103] On days 14 and 28, compared with the control group, HS in the brain showed a statistically significant dose-dependent decrease in all treatment groups (day 14: -31% for G3, -61% for G4, and -80% for G5 compared with the control group; day 28: -32% for G3, -32% for G4, and -67% for G5 compared with the control group). On day 14, HS in cerebrospinal fluid showed a statistically significant dose-dependent decrease in all treatment groups relative to the control group, but this decrease was not significant on day 28 (day 14: -45% for G3, -62% for G4, and -57% for G5 relative to the control group; day 28: -15% for G3, -23% for G4, and -8% for G5 relative to the control group).

[0104] In other words, the GC1130A treatment group (≥1.5 μg / dose) was shown to have dose-dependent HS depletion in the brain, which was generally observed up to 28 days post-mortem, while in the CSF, significant HS depletion was observed only on day 14 in the GC1130A treatment group with 1.5 μg / dose.

[0105] In summary, the effects on the brain and cerebrospinal fluid were determined based on the effective concentration of GC1130A and the analysis of the maximum volume in the mouse ventricles (5 μL / dose), at the lowest dose of 1.5 μg / dose.

[0106] Furthermore, strong correlations in hourly and overall analyses confirmed a significant correlation between HS levels in the brain and CSF in response to GC1130A administration (Figure 3).

[0107] Example 3: Repeated-dose efficacy study of GC1130A

[0108] The purpose of this study was to evaluate the efficacy of GC1130A administered via ICV for the treatment of central nervous system symptoms in MPS IIIA mice.

[0109] A formulation containing GC1130A protein (5 mM histidine, 125 mM NaCl, 1.8% trehalose, 0.005% PS20, pH 8.11) was used, and an MPS IIIA mouse model (C57BL / 6 Sgshmps3a gene WT, homozygous mutant, 2 weeks old, male & female) was used, and each mouse group is shown in Table 3 below.

[0110] [Table 3]

[0111]

[0112] Instructions for Use 10 / 16 pages 12 CN 122121891 A

[0113] Cerebrospinal fluid (CSF) samples were collected during autopsy, and the isolated brain was stored in a low-temperature freezer until HS measurement was performed.

[0114] HS levels in mouse CSF and brain samples were measured by LC-MS / MS, and biochemical markers, namely open field test behavioral assessment, were performed to verify the improvement in central nervous system function. In addition, MRI analysis and pathological analysis were performed by immunohistochemistry to confirm changes in actual brain structure (Table 4).

[0115] [Table 4]

[0116]

[0117] The biochemical parameter HS in both brain and CSF of the GC1130A 15-60 μg treatment group was significantly reduced, and the degree of HS inhibition in the brain was greater than that in CSF. Direct injection into the ventricles of the brain is likely the reason for these results. A 2-week interval administration produced a greater inhibitory effect than a 4-week interval administration, but HS was also effectively inhibited at the 4-week interval, and no dose-dependent or sex differences were observed (Figure 4).

[0118] Furthermore, a significant positive correlation was found between CSF and HS content in brain tissue (Figure 5). Specification 11 / 16 pages 13 CN 122121891 A

[0119] Open field behavior experiments confirmed that all behavioral parameters were improved in the treatment group with repeated administration of GC1130A. To ensure statistical significance, data from the carrier treatment groups in WT and MPS IIIA mice were calculated by adding the treatment groups every two weeks (Q2W) and once a month (Q4W), and no statistical difference was found between the two groups. Compared with the control group, the total activity of GC1130A showed a trend of increasing activity (Figure 6).

[0120] The minimum dose for expected cognitive improvement was found to be 30 μg / dose x 1 / month. Even if HS in the CSF is reduced by 50% or more, cognitive improvement cannot be expected if the CSF HS is below the level of carrier therapy for 2 weeks.For cognitive improvement, CSF HS must be maintained at 50% or less of the carrier for at least 2 weeks (15 μg / dose x 2 / month).

[0121] As a result, HNS was found to be effective against MPS IIIA in mice when administered at a dose of 15-60 μg / dose every 2 to 4 weeks.

[0122] Example 4: Biodistribution of GC1130A conjugated with fluorescent dye in mice after IV or ICV administration

[0123] The aim of this study was to analyze the pharmacokinetics of GC1130A in mice according to the route of administration and to assess the distribution profile of GC1130A in different tissues over time.

[0124] After a single dose of 10 mg / kg of GC1130A conjugated with fluorescent dye was administered to mice via IV or ICV, fluorescence intensity was imaged at time curves of 0.083, 1, 2, 4, 8, 24, 48, 96, and 192 hours using an in vivo imaging system (IVIS) to determine the distribution by organ (Table 5).

[0125] [Table 5]

[0126]

[0127] When GC1130A was administered via IV, higher fluorescence intensity was observed in the rest of the body compared to the brain, while when administered via ICV, higher fluorescence intensity was observed in the brain compared to other organs (Fig. 7A).

[0128] Fig. 7B shows the mean radioefficiency-time curves of the brain after IV or ICV administration of GC1130A. The mean radioefficiency in the brain was the highest after ICV injection compared to other administration regimens.

[0129] Example 5: Pharmacokinetics of GC1130A in the mouse brain after ICV administration

[0130] This study aimed to evaluate the pharmacokinetics of GC1130A in the brain of 5-week-old C57BL / 6 mice after a single ICV administration.

[0131] After ICV administration to 5-week-old C57BL / 6 mice, four mice were necropsed and the brains were homogenized for each of the following time curves: 0.25, 0.5, 1, 2, 4, 8, 24, 48, 96, 168, 240, 336, 504, and 672 hours (Table 6). The homogenates were used to measure the concentration of GC1130A in the brain by digital ELISA assay and pharmacokinetic analysis was performed.

[0132] [Table 6] Product Specification 12 / 16 pages 14 CN 122121891 A

[0133]

[0134] Dose-dependent drug exposure was observed in the brain after a single ICV administration of GC1130A, with a half-life of approximately 7 days (Figure 8).

[0135] Example 6: Quantitative analysis of GC1130A in the brain based on repeated ICV administration

[0136] After autopsy, the brain tissue of one male and one female in each group was embedded in paraffin.After immunostaining using a staining device, the stained slides were scanned to establish regions of interest (ROIs), and the expression of each biomarker was quantified.

[0137] After repeated ICV administration of GC1130A, brain pathological changes were quantitatively analyzed to confirm the drug delivery and efficacy via the CNS through brain administration (Figure 9).

[0138] Example 7: Quantitative IHC analysis of brain pathological changes after repeated ICV administration - LAMP2, CD68 analysis

[0139] After autopsy, brain tissue from one male and one female in each group was embedded in paraffin. After immunostaining using a staining device, the stained slides were scanned to establish regions of interest (ROIs), and the expression of each biomarker was quantified. Instructions for Use, pages 13 / 16, CN 122121891 A

[0140] Markers of inflammation and microglia, namely LAMP2 and CD68, were found to be increased in the MPS IIIA vector group and decreased in the GC1130A group (Figures 10 and 11). Furthermore, these markers were observed to decrease more at Q2W (2-week interval) than at Q4W (4-week interval). GC1130A was also detected in the same brain regions where LAMP2 and CD68 markers were decreased.

[0141] Example 8: Analysis of GC1130A Dosage in Humans

[0142] Example 8-1: Determination of Starting Dose of GC1130A in Pediatric Clinical Practice

[0143] The pediatric dose recommended in this example is based on the method validated in Hammon K, et al, Clin Transl Sci, 14, 1810-1821, 2021, and used to translate non-clinical data into human applications for ultra-rare neurodegenerative pediatric diseases. Allometric growth scaling and PK modeling methods were developed to predict the dose from non-clinical data. These methods were compared and analyzed to select the most conservative value as the clinical starting dose.

[0144] Example 8-2: Calculation of Human Equivalent Dose (HED) Based on Brain Weight

[0145] The principle of allometric growth scaling is based on direct administration to body parts and is usually based on body surface area, but in the case of GC1130A, it is a scaling based on the proportion of brain weight, as it is administered directly to brain tissue via intraventricular administration.

[0146] Data on mouse brain weight were obtained from repeated-dose in vivo efficacy studies in MPSIIIA mouse models. Treatment groups in this study received 15, 30, and 60 μg, administered every 2 weeks. All treatment groups showed significant efficacy, but no significant dose-response was observed. To calculate the human-to-animal brain weight ratio, the mean mouse brain weight obtained from previous studies was 0.4662 g. The mean monkey brain weight, obtained from a 28-week safety study, was 70 g.The brain weight is consistent with reported brain weights in juvenile monkeys. The estimate of human brain weight was obtained from a paper studying the relationship between brain weight and body weight with age. Brain weights for men and women were used as averages for each age category (Table 7).

[0147] [Table 7]

[0148] Human brain to animal brain weight ratio

[0149] The brain weight ratio was multiplied by the dose used in the mouse efficacy study and the dose used in the monkey safety study to obtain the HED (Table 8). Specification 14 / 16 pages 16 CN 122121891 A

[0150] [Table 8]

[0151]

[0152]

[0153] Human ICV dose based on brain weight

[0154] The therapeutic index based on MTD (maximum tolerated dose) and MED (minimum effective dose) can be used to calculate MRSD (maximum recommended starting dose), which is the maximum dose recommended in the initial clinical trial. However, due to the nature of biopharmaceuticals, no toxicity has been determined in the specification (pages 15 / 16, CN 122121891 A), so the maximum dosable dose was selected as the NOAEL (No Observed Adverse Effect Level) of the drug. Considering safety factors, 1 / 10 of the NOAEL was used as the standard for the clinical starting dose.

[0155] Since the selection was based on the clinical starting dose NOAEL, the NOAEL was converted to a human dose and divided by a safety factor of 10, which equals 23 mg.

[0156] A low single-dose efficacy study (Example 2) confirmed the effects on the brain and cerebrospinal fluid when administered to mice at a dose of 1.5 g / dose, which translates to a human dose of 3.5 mg (1 / 10 of the human dose of 35 mg in the 15 µg group in Table 8). This indicates that 23 mg is a safe and effective dose.

[0157] Examples 8-3: Selection of the clinical starting dose in a pediatric population

[0158] Since GC1130A is administered directly into the brain tissue via intraventricular administration, it is scaled not only by brain weight but also by cerebrospinal fluid volume. The average weight of a cynomolgus macaque is 3.6 kg and its average cerebrospinal fluid volume is 11.6 mL. A 2-year-old child has an average weight of 14.0 kg and an average cerebrospinal fluid volume of 3 to 4 mL / kg, approximately 42 to 56 mL.The ratio of cerebrospinal fluid volume between monkeys and humans is approximately 1:4 (William Bonadio, J Emerg Med. 2014 Jan;46(1): 141-50, Cheryl D Fryar, et al, Natl Health Stat Report. 2021 Aug:(160):1-24, Jenna M Sullivan, et al, J Transl Med. 2020 Aug 8;18(1):309).

[0159] For GC1130A, the NOAEL in a 28-week repeated-dose monkey safety study was 15 mg, which, when multiplied by four, yielded a human dose of 60 mg. Dividing this converted value by a safety factor of 10 yielded 6 mg.

[0160] Industrial Applicability

[0161] This invention relates to the optimal dosage and cycle of heparan-N-sulfatase (HNS), which effectively reduces the accumulation of heparan sulfate (HS), while improving patient cognition, and can be used as an enzyme replacement therapy (ERT) for the treatment of Santa Felipe syndrome type A (MPS IIIA).

[0162] Although certain aspects of the invention have been described in detail above, it will be apparent to those skilled in the art that these specific descriptions are merely preferred embodiments and are not intended to limit the scope of the invention. Therefore, the essential scope of the invention will be defined by the appended claims and their equivalents.

[0163] Sequence Listing Free Text

[0164] Accompanying electronic document. Instruction manual, page 16 / 16, 18 CN 122121891 A, Figure 1; Instruction manual, Figure 1 / 10, page 19 CN 122121891 A, Figure 2; Instruction manual, Figure 2 / 10, page 20 CN 122121891 A, Figure 3; Instruction manual, Figure 3 / 10, page 21 CN 122121891 A, Figure 4; Figure 5; Instruction manual, Figure 4 / 10, page 22 CN 122121891 A, Figure 6a; Instruction manual, Figure 5 / 10, page 23 CN 122121891 A, Figure 6b; Instruction manual, Figure 6 / 10, page 24 CN 122121891 A, Figure 7; Instruction manual, Figure 7 / 10, page 25 CN 122121891 A, Figure 8; Figure 9; Instruction manual, Figure 8 / 10, page 26 CN 122121891 A, Figure 10; Instruction manual, Figure 9 / 10, page 27 CN 122121891 A Figure 11. Appendix to the instruction manual, page 10 / 10, CN 122121891 A.

Claims

1. A pharmaceutical composition for the prevention or treatment of San Phillips syndrome type A (MPS IIIA), comprising heparin N-sulfatase (HNS), in, The heparin N-sulfatase is administered to patients at doses of 3 to 150 mg per dose, at intervals of 2 to 4 weeks, via intraventricular injection (ICV), intracerebral injection (IC), or intrathecal injection (IT).

2. The pharmaceutical composition according to claim 1, wherein, The heparin N-sulfatase is administered at a dose of 6 to 100 mg per dose.

3. The pharmaceutical composition according to claim 1, wherein, The total volume of each dose is 10 mL or less.

4. The pharmaceutical composition according to claim 1, wherein, The pharmaceutical composition further comprises 1 to 40 mM histidine buffer.

5. The pharmaceutical composition according to claim 1, wherein, The pharmaceutical composition further includes sugars.

6. The pharmaceutical composition according to claim 5, wherein, The sugar is at least one selected from the group consisting of trehalose, sucrose, maltose, lactose, and sorbitol.

7. The pharmaceutical composition according to claim 6, wherein, The concentration of the sugars included is from 0.1 to 5.0 w / v.

8. The pharmaceutical composition according to claim 1, wherein, The composition further comprises a salt.

9. The pharmaceutical composition according to claim 8, wherein, The salt is NaCl or KCl.

10. The pharmaceutical composition according to claim 9, wherein, The concentration of the salts included is from 30 mM to 500 mM.

11. The pharmaceutical composition according to claim 1, wherein, The composition further includes a surfactant.

12. The pharmaceutical composition according to claim 11, wherein, The surfactant is polysorbate 20 or polysorbate 80.

13. The pharmaceutical composition according to claim 12, wherein, The concentration of the surfactant included is from 0.0001 to 0.1 w / v.

14. The pharmaceutical composition according to claim 1, wherein, It is administered to the central nervous system via intraventricular injection (ICV).

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