Pharmaceutical compositions for treating acid sphingomyelinase deficiency

Abstract

To provide compositions of recombinant human ASM (rhASM) for treating acid sphingomyelinase deficiency (ASMD).SOLUTION: A composition comprises a recombinant human acid sphingomyelinase, sodium phosphate, methionine, and sucrose.SELECTED DRAWING: None

Description

[Background technology]

[0001] Acid sphingomyelinase deficiency (ASMD) is a rare, life-threatening lysosomal storage disorder. It is an autosomal recessive genetic disease caused by mutations in the SMPD1 gene, which encodes the lysosomal enzyme acid sphingomyelinase (ASM) (Non-Patent Document 1). ASMD patients are unable to metabolize sphingomyelin, which results in its accumulation in the lysosomes of multiple organs and, in severe cases, leads to visceral disease and neurodegeneration. ASMD patients have increased cholesterol and other lipids in the spleen, liver, lungs and bone marrow.

[0002] Infantile neurovisceral ASMD (also known as Niemann-Pick disease type A or NPD A) is the most severe disease phenotype and is characterized as early onset and acute neuropathic. NPD A causes growth failure, hepatosplenomegaly, and rapidly progressive neurodegeneration. Patients die in infancy (Non-Patent Document 2).

[0003] Patients with chronic visceral ASMD (NPD B) and chronic neurovisceral ASMD (NPD A / B) develop symptoms from infancy through adulthood (Non-Patent Document 3; Non-Patent Document 4). NPD B patients are usually diagnosed in childhood, typically after age 2. Most NPD B patients live into adulthood. NPD A / B patients are classified as intermediate types that present with childhood neurological symptoms that may manifest as neurodegenerative diseases. Morbidity from liver, lung, and hematopoietic system disease occurs in all patients with chronic ASMD and includes hepatosplenomegaly, liver dysfunction, infiltrative lung disease, and thrombocytopenia (Non-Patent Document 5; Non-Patent Document 6). Childhood growth restriction and bone disorders such as low bone density are also common features of chronic ASMD (Non-Patent Document 7). The main causes of death in these patients are lung and liver disease (Non-Patent Document 8; Non-Patent Document 9).

[0004] Due to the high morbidity and mortality associated with ASMD, there is an urgent need for effective treatments for this genetic disease. [Prior art documents] [Non-patent literature]

[0005] [Non-Patent Document 1] Schuchman et al., Mol. Genet. Metab. 120(1-2):27-33 (2017) [Non-Patent Document 2] McGovern et al., Neurology 66(2):228-232 (2006) [Non-Patent Document 3] Wasserstein et al., Pediatrics 114(6):e672-677(2004) [Non-Patent Document 4] Wasserstein et al., J. Pediatr. 149(4):554-559 (2006) [Non-Patent Document 5] McGovern et al., Genet. Med. 15(8):618-623 (2013) [Non-Patent Document 6] McGovern et al., Orphanet J. Rare Dis. 12(1):41(2017) [Non-Patent Document 7] Wasserstein et al., J. Pediatr. 142(4):424-428 (2003) [Non-Patent Document 8] McGovern et al., Pediatrics 122(2):e341–349 (2008) [Non-Patent Document 9] Cassiman et al., Mol. Genet. Metab. 118(3):206-213 (2016) Summary of the Invention [Problem to be solved by the invention]

[0006] The present invention provides compositions of recombinant human ASM (rhASM) for treating ASMD. In some embodiments, the compositions include rhASM, sodium phosphate, methionine, and sucrose (or trehalose). In certain embodiments, the rhASM is oliprodase alpha (SEQ ID NO: 2). [Means for solving the problem]

[0007] In some embodiments, the composition is lyophilized. The lyophilized composition of the present invention can be, for example: 4-7% w / w oliprodase alfa, 3-7% w / w sodium phosphate, 15-25% w / w L-methionine, and It may contain 65-75% w / w sucrose.

[0008] In a particular embodiment, the lyophilized composition of the present invention comprises: 5.5% w / w oliprodase alfa, 2.3% w / w sodium phosphate dibasic heptahydrate, 2.6% w / w sodium phosphate monobasic monohydrate, 20.5% w / w L-methionine, and It may contain 68.6% w / w sucrose.

[0009] In some embodiments, the composition is an aqueous liquid composition. The aqueous liquid composition may, for example, comprise: 1-10mg / mL oliprodase alfa, 10-50 mM sodium phosphate, 70-150mM L-methionine, and It may contain 1-10% w / v sucrose, Here, the composition has a pH of 5 to 8.

[0010] In certain embodiments, the aqueous liquid composition of the present invention comprises: 3-5mg / mL oliprodase alfa, 10-30 mM sodium phosphate, 80-120mM L-methionine, and It may contain 4-6% w / v sucrose, Here, the composition has a pH of 6-7.

[0011] In certain embodiments, the aqueous liquid composition of the present invention comprises: 4mg / mL oliprodase alfa, 20mM Sodium Phosphate, 100mM L-methionine, and may contain 5% w / v sucrose, Here, the composition has a pH of 6.5.

[0012] In some embodiments, the aqueous liquid composition of the present invention may further comprise 0.005% w / v Polysorbate 80.

[0013] The invention further provides compositions obtainable by drying (e.g., freeze-drying or spray-drying) the aqueous liquid compositions described herein. The invention also provides a method for producing a freeze-dried composition comprising freeze-drying the aqueous liquid compositions described herein. do.

[0014] In some embodiments, the present invention provides a vial comprising a lyophilized composition described herein. In certain embodiments, the lyophilized composition in the vial comprises or consists essentially of: 21.2mg oliprodase alfa, 9.0 mg sodium phosphate dibasic heptahydrate, 10.0 mg sodium phosphate monobasic monohydrate, 79mg L-Methionine, and 265mg sucrose.

[0015] In some embodiments, the lyophilized composition is reconstituted in 5.1 mL of sterile water to obtain an aqueous liquid composition.

[0016] In certain embodiments, the lyophilized composition in the vial comprises or consists essentially of: 4.8mg oliprodase alfa, 2.0 mg sodium phosphate dibasic heptahydrate, 2.3mg sodium phosphate monobasic monohydrate, 17.9mg L-Methionine, and 60mg sucrose.

[0017] In some embodiments, the lyophilized composition is reconstituted in 1.1 mL of sterile water to obtain an aqueous liquid composition.

[0018] The invention further provides an article of manufacture comprising: 1) a vial containing a lyophilized composition described herein; and 2) a vial containing, for example, sterile water, 0.9% sodium chloride, or phosphate buffered saline for reconstituting the lyophilized composition.

[0019] The present invention further provides a method of treating ASMD in a human patient comprising administering to the patient a composition described herein, wherein the composition, if it is a lyophilized composition, is reconstituted to a liquid form prior to administration.

[0020] The present invention further provides the compositions described herein for use in the treatment of ASMD in a human patient.

[0021] The invention further provides the use of a composition described herein for the manufacture of a medicament for treating ASMD in a human patient.

[0022] In some embodiments, the treatment of ASMD described herein is for Niemann-Pick Type A / B or Type B, or a non-neurological symptom of ASMD. [Brief description of the drawings]

[0023] [Figure 1]FIG. 1A shows the stability of rhASM as measured by specific (enzymatic) activity after storage for 2 weeks at 30° C. in succinate, citrate, citrate / phosphate, or phosphate buffers of various pHs. FIG. 1B shows the stability of rhASM as measured by percent high molecular weight species (% HMWS) after storage for 1 week at 30° C. in succinate, citrate, citrate / phosphate, or phosphate buffers of various pHs. HMWS was determined by size exclusion chromatography (SEC). FIG. 1C shows the stability of rhASM as measured by thermal stability in citrate / phosphate or phosphate buffers at various pHs. Thermal stability was determined by differential scanning calorimetry. [Diagram 2] Figure 2A shows the specific activity of rhASM over time in 10 mM, 20 mM, 50 mM, or 100 mM phosphate buffer at pH 6.5 at 30° C. Figure 2B shows the physical stability of rhASM over time as measured by % HMWS in 10 mM, 20 mM, 50 mM, or 100 mM phosphate buffer at pH 6.5 at 30° C. [Diagram 3] Figure 3A shows the effect of 5% w / v mannitol, sucrose, or trehalose on the specific (enzymatic) activity of 4 mg / mL rhASM before (liquid) and after (lyo) lyophilization. Figure 3B shows the effect of 5% w / v mannitol, sucrose, and trehalose on the physical stability of 4 mg / mL rhASM as measured by %HMWS before (liquid) and after (lyo) lyophilization. [Figure 4] Figure 4A shows the specific activity of rhASM over time at 5°C. RhASM was lyophilized from solutions containing 5% mannitol, 5% sucrose, or 3% mannitol and 2% sucrose (all w / v concentrations). Figure 4B shows the physical stability of rhASM over time at 5°C as measured by % HMWS. RhASM was lyophilized from solutions containing 5% mannitol, 5% sucrose, or 3% mannitol and 2% sucrose (all w / v concentrations). [Diagram 5]Figure 5A shows the specific activity of rhASM over time at 5°C. RhASM was lyophilized from a solution containing 5% sucrose with or without 100 mM methionine (all w / v concentrations). Figure 5B shows the physical stability of rhASM over time at 5°C as measured by % HMWS. RhASM was lyophilized from a solution containing 5% w / v sucrose with or without 100 mM methionine (all w / v concentrations). [Figure 6] FIG. 6 shows the effect of pH, protein concentration, methionine concentration, and sucrose concentration on the percentage of dimer over time in liquid rhASM compositions at 2-8° C. [Figure 7] FIG. 7 shows the effect of pH, protein concentration, methionine concentration, and sucrose concentration on rhASM specific activity over time in a liquid composition at 2-8° C. [Figure 8] 8 shows % HMWS over time for liquid rhASM compositions with varying pH, protein concentration, methionine concentration, and sucrose concentration at 2-8° C. Formulation numbers are indicated to the right of the graph. [Figure 9] 9 shows % aggregation over time in liquid rhASM compositions at various pH, protein, methionine, and sucrose concentrations at 2-8° C. Formulation numbers are indicated to the right of the graph. [Figure 10] FIG. 10 shows % dimer, % aggregation, and specific activity at 25° C. in liquid rhASM compositions at various pHs. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0024] The present invention provides a composition comprising recombinant human ASM, e.g., oliprodase alpha, and one or more pharma- ceutically acceptable excipients. The compositions of the present disclosure have improved stability and shelf life compared to other compositions. In some embodiments, the compositions of the present invention are pharmaceutical compositions, i.e., compositions that are in such a form or can be prepared in such a form that the biological activity of the active ingredient is effective while not containing additional ingredients that are significantly toxic or otherwise cause undesirable side effects in patients unrelated to the active ingredient. The terms "pharmaceutical composition" and "pharmaceutical formulation" are used interchangeably herein. The pharmaceutical compositions of the present invention are useful for treating patients with ASM deficiency, as further described below.

[0025] Recombinant Human Acid Sphingomyelinase ASM is an enzyme that catalyzes the degradation of sphingomyelin to ceramide and phosphorylcholine. "Recombinant human ASM" refers to a human ASM that is prepared by recombinant means and has a distinct homology to the wild-type sequence. It refers to human ASM with or without specific amino acid modifications. For example, recombinant human ASM is expressed in cultured mammalian host cells (such as COS, CHO, HeLa, 3T3, 293T, NS0, SP2 / 0, or HuT 78 cells) or in animals transgenic for the human ASM coding sequence.

[0026] In some embodiments, the recombinant human ASM is oributidase alpha. Oributidase alpha is the glycoform alpha of human ASM (EC-3.1.4.12) produced in CHO cells. Mature oributidase alpha is a 570 amino acid polypeptide that retains the enzymatic and lysosomal targeting activity of the native human protein. The amino acid sequence of oributidase alpha, including the leader sequence (residues 1-57), is shown below as SEQ ID NO:1, where the leader sequence is in italics and bold. The mature oributidase alpha sequence (SEQ ID NO:2 spanning residues 58-627 of SEQ ID NO:1) is lacking the leader sequence.

[0027] [ka]

[0028] In other embodiments, human ASM useful in the invention is 99%, 98%, 97%, 96%, or 95% identical in amino acid sequence to oligopeptidase alpha. For example, the human ASM in the composition can have the sequence set forth in U.S. Patent No. 6,541,218, the disclosure of which is incorporated herein in its entirety. The sequence (SEQ ID NO:3) is shown below with the leader sequence (residues 1-59) in italics and bold, where the mature protein (SEQ ID NO:4, spanning residues 60-629 of SEQ ID NO:3) is absent the leader sequence.

[0029] [ka]

[0030] The human ASM in the composition may also be identical in amino acid sequence to the human ASM disclosed in the UNIPROT database as sequence P17405-1 or a polymorphic variant thereof. The P17405-1 sequence is shown below (SEQ ID NO:5) with the leader sequence (residues 1-59) in italics and bold, whereas the mature protein (SEQ ID NO:6 spanning residues 60-629 of SEQ ID NO:5) is absent the leader sequence.

[0031] [ka]

[0032] Recombinant human acid sphingomyelinase compositions The compositions of the invention comprise recombinant human ASM and exhibit excellent stability with respect to the enzyme. "Stable" or "stability" refers to the ability of the active ingredient in the composition to retain its physical stability, chemical stability, and / or biological activity during storage and / or when subjected to physical or chemical stress. Stability can be at a selected temperature, e.g., under refrigerated conditions (e.g., 2-8°C), or at room temperature (e.g., 23-25°C), for a selected period of time, e.g., 16 weeks, 24 weeks, 36 weeks, 4 months, 6 months, 1 year, 2 years, 3 years, or more. Protein stability is measured in assays that are performed within a shorter period of time, but whose results are indicative of stability in a clinical setting. Such assays include freeze / thaw assays, in which the protein composition is subjected to one or more freeze-thaw cycles; or agitation assays, in which the protein composition is subjected to mechanical agitation treatment for a predetermined period of time. Protein stability can be determined by storing the protein composition at a designated storage temperature (e.g., 2-8°C) for a selected period of time and analyzing its structural and functional attributes, such as the extent of dimerization or aggregation (e.g., as measured by size-exclusion HPLC or protein gels), proteolysis (e.g., as measured by size-exclusion HPLC or protein gels), color change of the composition, clarity of the liquid composition, enzymatic activity, glycan content and composition, receptor binding affinity, methionine residual oxidation, and biological activity of the composition.

[0033] The compositions of the present invention include one or more pharma- ceutically acceptable excipients. "Excipient" refers to an inert substance used as a diluent, vehicle, carrier, preservative, binder, or stabilizer for the active ingredient of a drug. For example, the compositions may include a buffer, an isotonicity agent, and / or a stabilizer, such as an antioxidant. In some cases, one agent may serve more than one of these purposes. In some embodiments, the compositions of the present invention include recombinant human ASM, such as oliprodase alpha, a buffer, such as sodium phosphate or sodium citrate, a stabilizer, such as L-methionine, and a non-reducing sugar, such as sucrose or trehalose. Human ASM has improved stability due to the particular makeup of the composition.The compositions of the invention may be aqueous solutions or lyophilized preparations.

[0034] liquid composition In some embodiments, the composition is an aqueous liquid composition comprising 1-10 mg / mL (e.g., 3-5 mg / mL) rhASM (e.g., oliprodase alpha); 10-50 mM (e.g., 10-30 mM) sodium phosphate; 70-150 mM (e.g., 80-120 mM) methionine (e.g., L-methionine); and 1-10% (e.g., 4-6%) w / v sucrose or trehalose. The pH of the aqueous liquid composition may be 5-8 (e.g., 6-7).

[0035] In some embodiments, the aqueous liquid compositions do not contain detectable amounts of mannitol, the most readily used crystalline excipient, as this can significantly increase aggregation of human ASM during or after lyophilization of the aqueous liquid compositions described herein.

[0036] In some embodiments, the aqueous liquid composition comprises 0.004-0.008%, 0.005-0.007%, or 0.005% w / v of a surfactant. Exemplary surfactants include non-ionic surfactants such as polysorbates (e.g., polysorbate 20 and 80) and poloxamers (e.g., poloxamer 188). In certain embodiments, the aqueous liquid composition comprises 0.005% polysorbate 80. In some cases, the presence of a surfactant may help reduce the turbidity of the liquid composition.

[0037] In some embodiments, the aqueous liquid composition contains no more than 0.05, 0.01, or 0.005 mM of a chelating agent, such as EDTA and EGTA; in exemplary embodiments, the aqueous liquid composition does not contain detectable amounts of a chelating agent. In some cases, the presence of a chelating agent, for example at concentrations greater than 0.05 mM or 0.1 mM, can increase aggregation and decrease the stability of human ASM, especially after extended storage periods, such as 12-16 weeks, or under non-refrigerated conditions, for example at 25° C.

[0038] In some embodiments, the aqueous liquid composition can include 0-50 ppm (eg, 15-30 ppm) zinc, which is carried over from the manufacturing process or added exogenously, for example.

[0039] In a particular embodiment, the aqueous liquid composition comprises or consists essentially of 4 mg / mL oligopeptidase alpha, 20 mM sodium phosphate, 100 mM methionine, and 5% (w / v) sucrose, and has a pH of 6.5. The term "consists essentially of" means that the composition does not contain detectable amounts of other components, or contains only trace amounts of certain substances resulting from the protein manufacturing process, and that such substances do not affect the biological activity of the enzyme or are harmful to the human patient.

[0040] In some embodiments, the composition is an aqueous liquid composition comprising 1-20 mg / mL (e.g., 10 mg / mL) rhASM (e.g., olidocidase alpha) and 10-50 mM (e.g., 20 mM) sodium phosphate. In certain embodiments, the aqueous liquid composition further comprises methionine (e.g., L-methionine) and sucrose or trehalose. In certain embodiments, the aqueous liquid composition further comprises 80-120 mM (e.g., 100 mM) methionine and 4-6% (e.g., 5%) (w / v) sucrose. In certain embodiments, the aqueous liquid composition has a pH of 6.5.

[0041] In some embodiments, the composition comprises 1-50 mg / mL (e.g., 3.8, 18, or 49 mg / mL) rhASM (e.g., oliprodase alpha) and 10-50 mg / mL The aqueous liquid composition comprises 1 mM (e.g., 20 mM) sodium phosphate. In certain embodiments, the aqueous liquid composition further comprises 1-15% (e.g., 5%, 6%, 7%, or 8%) sucrose or trehalose. In certain embodiments, the aqueous liquid composition further comprises 80-120 mM (e.g., 100 mM) methionine. In certain embodiments, the aqueous liquid composition has a pH of 6.5. The composition may comprise, for example, 3.8 mg / mL rhASM, 20 mM sodium phosphate, and 5% sucrose; 18 mg / mL rhASM, 20 mM sodium phosphate, and 5% sucrose; or 49 mg / mL rhASM, 20 mM phosphate, and 8% sucrose.

[0042] Aqueous liquid compositions can be prepared by mixing human ASM produced by recombinant techniques and subsequently purified from host cells with an excipient as described herein in water and adjusting the resulting mixture to the desired pH. For example, the human ASM and the desired excipients are added or buffer exchanged into a sodium phosphate buffer having a desired sodium phosphate concentration and pH.

[0043] In some embodiments, aqueous liquid compositions can be prepared by reconstituting a lyophilized composition of the invention, as described in more detail below, with a pharma- ceutically acceptable liquid, such as sterile water, saline (e.g., 0.9% sodium chloride), or phosphate-buffered saline.

[0044] Freeze-dried composition The present invention also provides a lyophilized composition. Such a composition can be prepared by freeze-drying the aqueous liquid composition described herein. The lyophilized composition is suitable for long-term storage. Freeze-drying can be carried out according to methods known in the art. For example, the liquid composition can be cooled to a subzero (Celsius) temperature (e.g., -5°C to -80°C) where freezing occurs, and then placed in a low pressure (partial vacuum) chamber to allow sublimation to occur (primary drying); here, the temperature of the composition can be increased in a second stage of drying (secondary drying) if necessary to further remove unwanted water molecules. In some embodiments, after completion of the lyophilization process, an inert gas such as nitrogen is introduced into the container of the composition (e.g., a glass vial) before sealing the container.

[0045] In some embodiments, the present invention provides a powder composition that can be prepared by, for example, spray drying the aqueous liquid composition described herein. The spray-dried composition is suitable for long-term storage. Spray drying can be carried out according to methods known in the art. For example, the liquid composition can be forced to disperse as small droplets of controlled size through an atomizer or spray nozzle into a hot gas stream in a chamber, resulting in the liquid composition being rapidly dried into a powder. The dried powder can then be collected at the bottom of the drying chamber. Other drying methods for preparing the powder composition are also contemplated.

[0046] The inventors have unexpectedly discovered that sucrose (or trehalose) and methionine present in the amounts described herein provide superior results during lyophilization; the lyophilized product retains the stability of human ASM during storage while retaining the elegant cake. The human ASM in the lyophilized compositions of the invention can maintain its biological activity under refrigerated conditions (e.g., 0-10° C., 2-8° C., or 4° C.) for at least 4 months (e.g., at least 6 months, or at least 12 months) without aggregation.

[0047] In some embodiments, the compositions of the present invention comprise 4-50% oliprodase alpha, 3-7% sodium phosphate, and 45-90% sucrose (all w / w percent). It is a lyophilized pharmaceutical composition. In a particular embodiment, the lyophilized composition comprises 5.5% oliprodase alpha, 20.6% L-methionine, 2.3% sodium phosphate dibasic heptahydrate, 2.6% sodium phosphate monobasic monohydrate, and 69.0% sucrose (all w / w percent). In a particular embodiment, the lyophilized composition comprises 6.6% oliprodase alpha, 3.0% sodium phosphate dibasic heptahydrate, 3.3% sodium phosphate monobasic monohydrate, and 87.1% sucrose (all w / w percent). In a particular embodiment, the lyophilized composition comprises 25.2% oliprodase alpha, 2.4% sodium phosphate dibasic heptahydrate, 2.6% sodium phosphate monobasic monohydrate, and 69.9% sucrose (all w / w percent). In a particular embodiment, the lyophilized composition comprises 47.8% oliprodase alpha, 1.7% sodium phosphate dibasic heptahydrate, 1.8% sodium phosphate monobasic monohydrate, and 48.8% sucrose (all w / w percentages).

[0048] In some embodiments, the compositions of the present invention are lyophilized pharmaceutical compositions comprising 4-7% oliprodase alpha, 15-25% L-methionine, 3-7% sodium phosphate, and 65-75% sucrose (all w / w percent). In certain embodiments, the lyophilized composition comprises 5.5% oliprodase alpha, 20.5% L-methionine, 2.3% sodium phosphate dibasic heptahydrate, 2.6% sodium phosphate monobasic monohydrate, and 68.6% sucrose (all w / w percent). In certain embodiments, the lyophilized composition may also comprise, for example, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, or 1.0% water.

[0049] In some embodiments, the present invention provides a vial containing a lyophilized pharmaceutical composition comprising 15-25 mg olivodase alpha, 75-85 mg L-methionine, 15-25 mg sodium phosphate, and 250-300 mg sucrose. Prior to use, the composition can be reconstituted with 4-6 mL of sterile water.

[0050] In some embodiments, the vial contains a lyophilized pharmaceutical composition comprising or consisting of 21.2 mg, 20.1 mg, 95.4 mg, or 259.7 mg of oliprodase alpha; 9.0 mg sodium phosphate dibasic heptahydrate; 10.0 mg sodium phosphate monobasic monohydrate; and 265 mg sucrose. The lyophilized composition may optionally contain 79.1 mg L-methionine. The lyophilized pharmaceutical composition may optionally contain 0-0.3 mg (e.g., 0.08-0.16 mg) of zinc, which may be carried over from the manufacturing process or added exogenously, for example. In certain embodiments, the vial may have a sterile nitrogen-filled atmosphere inside. In certain embodiments, the lyophilized composition may be reconstituted with 5.1 mL of sterile water to obtain a concentration of about 4.0 mg / mL, 3.8 mg / mL, 18 mg / mL, or 49 mg / mL of oliprodase alpha, respectively. The reconstituted composition is further diluted to a specific volume with 0.9% sodium chloride solution based on the dose to be administered.

[0051] In a particular embodiment, the vial contains a lyophilized pharmaceutical composition comprising or consisting of 21.2 mg oliprodase alpha, 79 mg L-methionine, 9.0 mg sodium phosphate dibasic heptahydrate, 10.0 mg sodium phosphate monobasic monohydrate, and 265 mg sucrose. The lyophilized pharmaceutical composition may optionally contain 0-0.3 mg (e.g., 0.08-0.16 mg) zinc, which may be carried over from the manufacturing process or added exogenously, for example. In a particular embodiment, the lyophilized pharmaceutical composition is in the form of a cake or a lyophilized powder. In a particular embodiment, the vial may have a sterile nitrogen-filled atmosphere inside. In a particular embodiment, the lyophilized composition is reconstituted in 5.1 mL of sterile water to obtain an oliprodase alpha concentration of about 4.0 mg / mL. The reconstituted composition may be added to a particular volume, depending on the dose to be administered. It is further diluted with 0.9% sodium chloride solution based on the above.

[0052] In some embodiments, the present invention provides a vial containing a lyophilized pharmaceutical composition comprising 3-5 mg olivopridase alpha, 15-17 mg L-methionine, 3-5 mg sodium phosphate, and 50-60 mg sucrose. Prior to use, the composition may be reconstituted in 0.8-1.2 mL of sterile water.

[0053] In a particular embodiment, the vial contains a lyophilized pharmaceutical composition comprising or consisting of 4.8 mg oliprodase alpha, 17.9 mg L-methionine, 2.0 mg sodium phosphate dibasic heptahydrate, 2.3 mg sodium phosphate monobasic monohydrate, and 60 mg sucrose. In a particular embodiment, the lyophilized pharmaceutical composition is in the form of a cake or a lyophilized powder. The lyophilized composition may optionally contain 0-0.06 mg zinc, which may be carried over from the manufacturing process or added exogenously, for example. In a particular embodiment, the vial may have a sterile nitrogen-filled atmosphere inside. In a particular embodiment, the lyophilized composition may be reconstituted in 1.1 mL of sterile water to obtain an oliprodase alpha concentration of about 4.0 mg / mL. The reconstituted composition is further diluted with 0.9% sodium chloride solution to a particular volume based on the dose to be administered.

[0054] manufactured goods The compositions of the present invention can be provided in an article of manufacture (e.g., a kit) that includes instructions for use in treating ASM disorders, and optionally other therapeutic agents. The pharma- ceutical active ingredient (e.g., rhASM) in the product can be provided in an amount that can be easily administered according to the dosing schedule described herein. For example, a "starter kit" can include multiple vials of rhASM in various amounts for use in a dose escalation regimen.

[0055] For example, the article of manufacture may include a vial containing 15-25 mg lipidase alpha, 75-85 mg L-methionine, 15-25 mg sodium phosphate, and 250-300 mg sucrose. In a particular embodiment, the article of manufacture provides a lyophilized composition containing 21.2 mg lipidase alpha, 79 mg methionine, 9.0 mg sodium phosphate dibasic heptahydrate, 10.0 mg sodium phosphate monobasic monohydrate, and 265 mg sucrose.

[0056] As another example, the article of manufacture may include a vial containing 3-5 mg oliprodase alpha, 15-17 mg L-methionine, 3-5 mg sodium phosphate, and 50-60 mg sucrose. In a particular embodiment, the article of manufacture provides a lyophilized composition containing 4.8 mg oliprodase alpha, 17.9 mg L-methionine, 2.0 mg sodium phosphate dibasic heptahydrate, 2.3 mg sodium phosphate monobasic monohydrate, and 60 mg sucrose.

[0057] In some embodiments, the article of manufacture may further include solutions for reconstituting the lyophilized composition and / or for further diluting the reconstituted composition prior to administration to a patient (e.g., sterile water, 0.9% sodium chloride, and / or phosphate buffered saline).

[0058] Uses of Acid Sphingomyelinase Compositions The pharmaceutical composition of the present invention can be administered parenterally to a patient in need thereof as an enzyme replacement therapy. "Parenteral administration" refers to a means of administration other than enteral and topical administration, usually by injection. Parenteral administration includes, but is not limited to, intravenous infusion or injection, as well as intramuscular, intradermal, intraperitoneal, and subcutaneous injections. In certain embodiments, the pharmaceutical composition The substance is administered via intravenous infusion.

[0059] Appropriate dosage levels of the pharmaceutical compositions described herein can be determined based on a variety of factors, including the patient's age, weight, medical condition, general health, and medical history, as well as the route and frequency of drug administration, the pharmacodynamics and pharmacokinetics of the ASM active ingredient in the drug, and any other drugs the patient may be taking concomitantly. In some embodiments, the pharmaceutical compositions described herein can be administered according to the dosage regimens described, for example, in U.S. Pat. No. 9,655,954 (Schuchman et al.). For example, a patient can receive increasing doses of human ASM, with dose strengths starting at, for example, 0.1 mg / kg or less and ending at 3 mg / kg (maintenance dose) or less, depending on the patient's age and condition. In some embodiments, the first one or two doses may be given at a dose strength of 0.03 mg / kg or 0.1 mg / kg for pediatric patients, or 0.1 mg / kg for adult patients; after the patient receives one or two doses at 0.03 and / or 0.1 mg / kg, the patient is then given successive doses of 0.3 mg / kg, 0.3 mg / kg, 0.6 mg / kg, 0.6 mg / kg, 1.0 mg / kg, 2.0 mg / kg, and 3.0 mg / kg. In certain embodiments, any of the doses may be repeated (e.g., doses at 1.0 mg / kg and 2.0 mg / kg). For some patients, a dose strength of 3.0 mg / kg is suitable as a maintenance dose, while for other patients, a lower dose strength may be sufficient for maintenance. The interval between successive doses may be 2 weeks, or shorter or longer than 2 weeks, as determined by the clinician as appropriate.

[0060] The present invention provides a method of using the pharmaceutical composition described herein for treating ASMD in a patient in need thereof, a pharmaceutical composition described herein for use in treating ASMD in a patient in need thereof, and a use of the pharmaceutical composition described herein for the manufacture of a medicament for treating ASMD in a patient in need thereof. In some embodiments, the pharmaceutical composition may be a lyophilized composition, which is reconstituted in a pharma- ceutical acceptable liquid, such as sterile water, 0.9% sodium chloride solution, or phosphate buffered saline.

[0061] The patient may be an adult (e.g., a patient aged 18 years or older, including geriatric patients aged 65 years or older). The patient may be a pediatric patient (a patient younger than 18 years, e.g., a patient aged newborn to 6 years, 6 to 12 years, or 12 to 18 years). In some embodiments, the patient may have NPD A / B or NPD B. In some embodiments, the patient may have NPD A. In certain embodiments, the pharmaceutical composition is for treating an adult or pediatric patient with chronic visceral ASMD (NPD B). In certain embodiments, the pharmaceutical composition is for treating a non-neurological symptom of ASMD in an adult or pediatric patient.

[0062] Exemplary embodiments Further specific embodiments of the present invention are described below. 1. A composition comprising recombinant human acid sphingomyelinase, sodium phosphate, methionine, and sucrose. 2. The composition comprises: 4-7% w / w oliprodase alpha (SEQ ID NO: 2), 3-7% w / w sodium phosphate, 15-25% w / w L-methionine, and 2. The composition of embodiment 1, which is a lyophilized composition comprising 65-75% w / w sucrose. 3. In essence: 5.5% w / w oliprodase alfa, 2.3% w / w sodium phosphate dibasic heptahydrate, 2.6% w / w sodium phosphate monobasic monohydrate, 20.5% w / w L-methionine, and The composition of embodiment 2, wherein the composition consists of 68.6% w / w sucrose. 4. The composition comprises: 1-10mg / mL oliprodase alfa, 10-50 mM sodium phosphate, 70-150mM L-methionine, and An aqueous liquid composition comprising 1-10% w / v sucrose, The composition of embodiment 1, wherein the composition has a pH of 5 to 8. 5. The composition comprises: 3-5mg / mL oliprodase alfa, 10-30 mM sodium phosphate, 80-120mM L-methionine, and An aqueous liquid composition comprising 4-6% w / v sucrose, The composition of embodiment 4, wherein the composition has a pH of 6-7. 6. In essence: 4mg / mL oliprodase alfa, 20mM Sodium Phosphate, 100mM L-methionine, and 5. The composition of embodiment 4, comprising 5% w / v sucrose, The composition has a pH of 6.5. 7. The composition of any one of embodiments 4-6, further comprising 0.005% w / v Polysorbate 80. 8. A composition obtained by freeze-drying the aqueous liquid composition of any one of embodiments 4 to 7. 9. A method for producing a lyophilized composition comprising: Obtaining an aqueous liquid composition according to any one of embodiments 4 to 7; and A method comprising lyophilizing the aqueous liquid composition. 10. In essence: 21.2mg oliprodase alfa, 9.0 mg sodium phosphate dibasic heptahydrate, 10.0 mg sodium phosphate monobasic monohydrate, 79mg L-Methionine, and A vial containing a lyophilized composition consisting of 265 mg sucrose. 11. In essence: 21.2mg oliprodase alfa, 9.0 mg sodium phosphate dibasic heptahydrate, 10.0 mg sodium phosphate monobasic monohydrate, 79mg L-Methionine, and 265m sucrose An aqueous liquid composition obtained by reconstituting a lyophilized composition consisting of the above in 5.1 mL of sterile water. 12. In essence: 4.8mg oliprodase alfa, 2.0 mg sodium phosphate dibasic heptahydrate, 2.3mg sodium phosphate monobasic monohydrate, 17.9mg L-Methionine, and A vial containing a lyophilized composition consisting of 60 mg sucrose. 13. In essence: 4.8mg oliprodase alfa, 2.0 mg sodium phosphate dibasic heptahydrate, 2.3mg sodium phosphate monobasic monohydrate, 17.9mg L-Methionine, and An aqueous liquid composition obtained by reconstituting a lyophilized composition consisting of 60 mg sucrose in 1.1 mL of sterile water. 14. An article of manufacture comprising a vial of embodiment 10 or 12 and a vial containing sterile water, 0.9% sodium chloride, or phosphate buffered saline for reconstituting the lyophilized composition. 15. A method of treating acid sphingomyelinase deficiency (ASMD) in a human patient, comprising administering to the patient a composition of any one of embodiments 1-8, 11, and 13, wherein the composition, if it is a lyophilized composition, is reconstituted to a liquid form prior to administration. 16. The composition of any one of embodiments 1-8, 11, and 13, for use in treating ASMD in a human patient. 17. Use of the composition of any one of embodiments 1 to 8, 11, and 13 for the manufacture of a medicament for treating ASMD in a human patient. 18. The method of embodiment 15, the composition for use of embodiment 16, or the use of embodiment 17, wherein ASMD is Niemann-Pick type A / B or type B. 19. The method, composition for use, or use of embodiment 18, wherein the treatment is for a non-neurological symptom of ASMD.

[0063] All publications and other references mentioned herein are incorporated by reference in their entirety. Although many documents are cited herein, this citation is not an admission that any of these documents form part of the general knowledge in the art. Unless otherwise defined herein, scientific and technical terms used in connection with the present invention shall have the meanings commonly understood by those of ordinary skill in the art. Exemplary methods and materials are described below, although methods and materials similar or equivalent to those described herein may also be used in practicing or testing the present invention. In case of conflict, the present specification, including definitions, will control. In general, the nomenclature used in connection with and techniques of cell and tissue culture, molecular biology, microbiology, genetics, analytical chemistry, synthetic organic chemistry, medicinal chemistry, and protein and nucleic acid chemistry described herein are well known and commonly used in the art. Enzymatic reactions and purification techniques are performed according to manufacturer's specifications as commonly accomplished in the art or as described herein. Furthermore, unless the context requires otherwise, the singular shall include the plural and the plural shall include the singular. Throughout this specification and the embodiments, the words "have" and "include" or variations such as "have," "having," "including," or "including" will be understood to mean the inclusion of a stated integer or group of integers but not the exclusion of other integers or groups of integers. EXAMPLES

[0064] In order that the invention may be better understood, the following examples are presented, which are for illustrative purposes only and should not be construed as limiting the scope of the invention in any manner. EXAMPLES

[0065] Recombinant human acid sphingomyelinase preparation This example describes a study evaluating the stability of various aqueous liquid and lyophilized compositions of oliprodase alpha.

[0066] material and method Solution turbidity The opalescence of the solutions was assessed by a spectroscopic turbidity assay. An optical density range of 340–360 nm was used to set the range for the opalescence category previously established based on the European Pharmacopoeia reference suspensions at specific NTU values. Analysis was performed on a SpectraMax Plus 384 microplate spectrophotometer (Molecular Devices, Sunnyvale, CA).

[0067] agglomeration Aggregation and dimer analysis was performed by SEC. Each sample was mixed by gentle pipette circulation five times before loading into the HPLC vial. SEC analysis was performed on an 1100 / 1200 series HPLC (Agilent, Santa Clara, CA) equipped with a TSK gel G3000SWXL (Tosoh Biosciences, Tokyo, Japan) analytical column and a corresponding guard column. The mobile phase used was 20 mM sodium phosphate, pH 6, 0.5 mL min -1 The run duration was 35 min at a flow rate of 200 mM sodium chloride. Three injections of approximately 80 μg were performed for each sample. Detection was performed by UV absorbance at 280 nm.

[0068] Levels of oliprodase alpha-related high molecular weight species (HMWS) were determined using SDS-PAGE under non-reducing conditions, followed by staining with Coomassie blue. An oliprodase alpha reference standard was included in each gel. Olipidase alpha samples were mixed with sample buffer and loaded onto a 4-20% Tris-Glycine gradient gel along with molecular weight markers. After electrophoresis at 125 V target for approximately 2 hours, the gel was stained with Coomassie blue and destained with methanol, acetic acid, and HPLC-grade water. Densitometric analysis was performed to provide quantitative results for the percentage of HMWS bands relative to all bands observed.

[0069] Enzyme activity rhASM samples were diluted 2000:1 in 1.2 mL library tubes. This procedure measured the rate of hydrolysis of 2-(N-hexadecanoylamino)-4-nitrophenylphosphorylcholine (HDA-PC) catalyzed by rhASM at 37 °C. The released chromophore was measured by absorbance at 415 nm using a SpectraMax Plus 384 microplate spectrophotometer. One unit of rhASM activity is defined as the amount of enzyme that produces 1 μmol of 2-(N-hexadecanoylamino)-4-nitrophenol per minute from HDA-PC under the specified assay conditions.

[0070] Protein concentration The protein concentration of rhASM samples was determined by absorbance at 280 nm. Samples were diluted 1:10 and 1:20 in duplicate using the corresponding buffer. Absorbance at 280 nm was measured on a SpectraMax Plus 384 microplate spectrophotometer.

[0071] pH pH analysis of samples was performed with a Thermo Electron Microprobe pH meter (Thermo Scientific, Beverly, MA). Thermo Orion 8203BN PerHecT Ross semi-micro glass probe (Thermo Scientific) was used.

[0072] Differential Scanning Calorimeter Differential scanning calorimetry (DSC) analysis was performed using a CAP-VP-DSC microcalorimeter (M The measurements were performed using a DSC analyzer (MicroCal-GE Healthcare, Northampton, MA). Samples were diluted to 0.4 mg / mL in the corresponding buffer. Samples were run from 15 to 100 °C at a scan rate of 200 °C / h. Data analysis was performed using the Origin 3D Scanner with DSC analysis add-on (MicroCal-GE Healthcare). The experiment was performed using OriginLab 7.0 (OriginLab, Northampton, MA).

[0073] result Buffers and pH Evaluation The effect of different buffer pHs and buffer types on the stability of rhASM was evaluated by incubating 4 mg / ml olivopridase alpha in 20 mM buffer at 30°C for 2 weeks and assessing the physical and functional stability of the enzyme (Figures 1A-1C).

[0074] Significant instability, both physical and functional, was observed in olivomycin alpha below pH 6.0. Enzyme activity decreased sharply below pH 6.0 but remained relatively constant above pH 6.0 (Figure 1A). Aggregation tendency was lowest between pH 5.5 and 6.5. A sharp increase in aggregation was observed below pH 5.5 and a gradual increase in aggregation above pH 6.5 (Figure 1B). At comparable pH values, stability was higher in phosphate buffer compared to citrate / phosphate buffer. These stability trends were consistent with data obtained from samples stored at refrigerated temperatures. The gradual increase in aggregation rate observed above pH 6.5 was supported by DSC analysis, which indicated decreased thermal stability at higher pH in phosphate buffer (Figure 1C). Based on these data, sodium phosphate buffer at pH 6.5 was identified as a suitable buffer system for rhASM formulations.

[0075] Next, the effect of ionic strength or buffer concentration on stability was investigated by varying the sodium phosphate concentration from 10 mM to 100 mM. As shown in Figures 2A and 2B, ionic strength at low buffer concentrations (10 mM, 20 mM, or 50 mM) had little effect on the enzymatic activity and physical stability of oliprodase alfa. The stability of oliprodase alfa in sodium phosphate, pH 6.5, was relatively consistent at buffer concentrations below 50 mM. However, there was a significant decrease in stability when the concentration of sodium phosphate was increased to 100 mM. Such concentrations resulted in a rapid increase in aggregated species and a concomitant decrease in activity (Figures 2A and 2B). The data at 30°C were consistent with those seen during storage of oliprodase alfa under refrigerated conditions. A sodium phosphate concentration of 20 mM was selected as the buffer concentration, which, although low, ensured sufficient buffering capacity.

[0076] Excipient evaluation Various pharma- ceutically acceptable excipients were evaluated for their effect on the stability of oliprodase alfa in liquid and lyophilized compositions. The effect of known stabilizers acting via preferential exclusion mechanisms on liquid stability was first investigated (e.g., Timasheff, Proc Nat Acad Sci USA. 99: 9721-9726 (2002); and Lee et al., J. Biol. Chem. 256: 7193-7201 (1981)). Sucrose, trehalose, and propylene glycol were added to a base formulation of 20 mM sodium phosphate, 0.005% PS80, pH 6.5, and the effect on the enzymatic activity and physical stability of 4 mg / mL oliprodase alfa was evaluated. Data showed that the presence of any of these preferential exclusion stabilizers did not enhance the stability of oliprodase alfa in the liquid state at 5°C or 30°C (data not shown). There was a loss of enzyme activity and physical stability under all conditions studied, although sucrose appeared to be slightly preferred over trehalose and propylene glycol.

[0077] Next, the effect of sucrose and trehalose on the stability of oliprodase alpha during freeze-drying was investigated. Mannitol was investigated because it is a bulking agent commonly used in freeze-drying (Figures 3A and 3B). The three polyols were included at 5% w / v in the liquid composition, which was then freeze-dried. The data in Figure 3A show that the addition of mannitol reduced the enzyme activity of oliprodase alpha after freeze-drying. The data in Figure 3B show that the addition of mannitol resulted in a significant increase in protein aggregation after freeze-drying. The increase in aggregation in the presence of sucrose and trehalose was minimal. As a cryoprotectant, sucrose was selected at a concentration of 5% w / v in the liquid composition, which could then be freeze-dried.

[0078] The stability of lyophilized oliprodase alfa formulations over time in the presence of mannitol was also examined. Olipudase alfa was lyophilized from a liquid composition containing 20 mM sodium phosphate buffer (pH 6.5), 0.005% PS80, and (i) 5% w / v mannitol, (ii) 5% w / v sucrose, or (iii) 3% mannitol and 2% sucrose. The data show that mannitol not only led to an immediate as well as a continuing increase in protein aggregation during lyophilization when used alone, but also when used in combination with sucrose for 6 months (Figures 3B and 4B). Enzyme activity at 6 months was also lower in the presence of mannitol than in its absence (Figure 4B). Thus, mannitol was deemed an inappropriate bulking agent for lyophilized rhASM formulations.

[0079] Since mannitol was found to be detrimental to the stability of rhASM during lyophilization, methionine was evaluated as a potential bulking agent. The enzymatic activity and physical stability of oliprodase alfa during liquid storage in the presence of L-methionine were evaluated and it was found that the addition of 100 mM L-methionine did not improve or adversely affect the liquid stability of oliprodase alfa at 5°C. As shown in Figures 5A and 5B, lyophilized compositions prepared from liquid compositions containing 100 mM L-methionine resulted in stable lyophilized constructs of oliprodase alfa. No changes in activity or aggregation were observed during 6 months of storage at 5°C. Furthermore, cakes obtained from lyophilized oliprodase alfa in the presence of methionine and sucrose had improved appearance versus those lyophilized with sucrose alone.

[0080] Identification of the appropriate amount of methionine for lyophilization was performed by X-ray diffraction (XRD) of oliprodase alfa cakes lyophilized from liquid compositions containing 20 mM sodium phosphate (pH 6.5), 5% w / v sucrose, and increasing amounts of L-methionine (data not shown). Samples without methionine were completely amorphous. There was some evidence of crystallization in samples containing 33 mM methionine, and further evidence of crystallization in samples containing 66 mM and 100 mM methionine. Lyophilization of oliprodase alfa at methionine levels below 100 mM (e.g., 10 mM and 33 mM) resulted in vials that appeared hollow and collapsed (shrinking). Therefore, 100 mM methionine was selected as the bulking agent for the lyophilized rhASM formulation. EXAMPLES

[0081] Robustness of Olipidase Alpha Composition To evaluate the robustness of compositions containing oligopeptidase alpha, sucrose, and L-methionine, each of these components was assayed at five different levels (low, medium-low, middle (control), medium-high, and high) compared to the control in 20 mM sodium phosphate buffer (Table 1).

[0082] [Table 1]

[0083] A total of 26 liquid formulation variants were generated (Table 2). Formulations 2 and 7 represent the control formulation or center point. The remaining 24 formulation variants represent various conditions around the center point. All 26 variants were stored at 2-8 °C (24 weeks or up to 12 months) and 25 °C (16 weeks) and their stability was monitored.

[0084] [Table 2]

[0085] At the end of the study period, the following parameters will indicate the stability of the compositions: (1) clear and colorless appearance; (2) pH 6.0-7.0; (3) aggregation ≤3.0%; (4) 3.5-4.5 mg / mL protein; (5) HMWS ≤15%; and (6) specific activity 11-42 U / mg. Figures 6-8 show the stability of the 26 formulation variants, respectively, after 24 weeks of storage at 2-8°C. Figure 1 shows the effect of different factors on % dimer, specific activity, and % HMWS in. The data show that there were no significant differences between the variants with respect to % dimer, specific activity, or % HMWS, and therefore all 24 variants were stable for 24 weeks at 2-8°C along with the two control formulations.

[0086] It was also observed that for some variants, there was no significant effect of varying pH and component concentrations on % dimer and specific activity through 36 weeks. Figure 9 shows that all variants had similar aggregation at 36 weeks.

[0087] At an accelerated temperature of 25°C, there was also little effect of excipients on stability over 16 weeks in the range tested. The most significant effect was observed for variants with different pH (Figure 10). At higher pH (6.8 to 7.0) there was more stability of olivomycin alfa, and at lower pH (below 6.5) there was more aggregation and dimerization.

[0088] This study shows that the variant formulation at 2-8 °C is robust and stable with the selected composition range.

Claims

1. 1. A vial for containing a lyophilized composition for treating acid sphingomyelinase deficiency (ASMD) in a human patient, the lyophilized composition comprising essentially about: 20mg oliprodase alfa, 5 mg disodium phosphate, 8 mg monosodium phosphate, 75 mg L-methionine, and 250 mg sucrose, Consists of: The vial.

2. 10. The vial of claim 1, wherein the lyophilized composition contains no more than 0.5% moisture, and optionally contains 0.3% moisture.

3. Essentially approximately: 20mg oliprodase alfa, 5 mg disodium phosphate, 8 mg monosodium phosphate, 75 mg L-methionine, and 250 mg sucrose, 2. An aqueous liquid composition for treating acid sphingomyelinase deficiency (ASMD) in a human patient, comprising:

4. 1. A vial for containing a lyophilized composition for treating acid sphingomyelinase deficiency (ASMD) in a human patient, the lyophilized composition comprising essentially: 4mg oripdase alfa, 0.9 mg disodium phosphate, 1.6 mg monosodium phosphate, 15 mg L-methionine, and 50 mg sucrose, The vial comprising:

5. 5. The vial of claim 4, wherein the lyophilized composition contains no more than 0.5% moisture, and optionally contains 0.3% moisture.

6. Essentially approximately: 4mg oripdase alfa, 0.9 mg disodium phosphate, 1.6 mg monosodium phosphate, 15 mg L-methionine, and 50 mg sucrose, 1. An aqueous liquid composition for treating acid sphingomyelinase deficiency (ASMD) in a human patient, comprising:

7. 10. An article of manufacture for treating acid sphingomyelinase deficiency (ASMD) in a human patient, comprising a vial of any one of claims 1, 2, 4, or 5, and a vial containing sterile water, 0.9% sodium chloride, or phosphate buffered saline for reconstituting the lyophilized composition.

8. 8. The article of manufacture of claim 7, wherein the lyophilized composition contains no more than 0.5% moisture, and optionally contains 0.3% moisture.

9. Use of a vial or composition according to any one of claims 1 to 6 for the manufacture of a medicament for treating Acid Sphingomyelinase Deficiency (ASMD) in a human patient.

10. 10. The use according to claim 9, wherein the ASMD is Niemann-Pick type A / B or Niemann-Pick type B.

11. 11. The use of claim 10, wherein the treatment is for a non-neurological symptom of ASMD.

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