Methods of treating congenital disorders of glycosylation (CDG) and ataxia in subjects suffering from cdg

Abstract

The disclosure provides phosphorylated carbohydrate replacement therapies (CRT) that include compositions of phosphorylated carbohydrates, such as mannose-1-phosphate (M1P), and phospholipids, as well as methods for preparing such compositions. Such compositions are suitable for pharmaceutical delivery of phosphorylated carbohydrates, such as M1P, for treating CDG type I and CDG type II diseases, including phosphomannomutase 2-congenital disorder of glycosylation (PMM2-CDG), as well as ataxia.

Description

PCT / US25 / 17678 27 February 2025 (27.02.2025)Attorney Docket No.: 74220-20015.40METHODS OF TREATING CONGENITAL DISORDERS OF GLYCOSYLATION (CDG) AND ATAXIA IN SUBJECTS SUFFERING FROM CDGCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to U.S. Provisional Patent Application No. 63 / 559,841, filed February 29, 2024, U.S. Provisional Patent Application No. 63 / 559,858, filed February 29, 2024, and U.S. Provisional Patent Application No. 63 / 559,845, filed February 29, 2024, each of which is incorporated herein by reference in its entirety.FIELD

[0002] The present invention relates generally to methods of treating congenital disorders of glycosylation (CDG), as well as methods of treating ataxia in a human suffering from CDG.BACKGROUND

[0003] Glycosylation, the enzymatic attachment of carbohydrates (glycans) to proteins and lipids, is a co-translational and post-translational modification (PTM) that is more common than any other PTM as it applies to a majority of proteins synthesized in the rough endoplasmic reticulum (ER). Glycosylation plays a critical role in a variety of biological processes of membrane and secreted proteins. In the ER, glycosylation defines the protein structure and folding and acts as a quality control mechanism that dictates the export of properly folded proteins to Golgi or targets misfolded ones for degradation. Glycan moieties may also act as ligands for cell surface receptors to mediate cell attachment or stimulate signal transduction pathways. Congenital disorders of glycosylation, also known as CDG syndromes, are a group of rare genetic diseases where tissue proteins and / or lipids carry defective glycosylation and / or lack of glycosylation. These diseases are linked to numerous enzymatic deficiencies and often times cause severe, sometimes fatal, impairments of the nervous system, muscles, intestines, and several other organ systems.

[0004] Common clinical symptoms in children with CDG include hypotonia, developmental delay, failure to thrive, hepatic dysfunction, coagulopathy, hypothyroidism, esotropia, abnormal fat pattern and inverted nipples, hypoglycemia, seizure, cerebellar hypoplasia, and stroke-like episodes in a developmentally delayed child. At an older age, in1 sf-6569471PCT / US25 / 17678 27 February 2025 (27.02.2025)Attorney Docket No.: 74220-20015.40 adolescence or adulthood, the presentation may include ataxia, cognitive impairment, the absence of puberty in females, small testes in males, retinitis pigmentosa, scoliosis, joint contractures, and peripheral neuropathy.

[0005] CDG may be classified into two groups: CDG type I and CDG type II. CDG type I is characterized by defects in the initial steps of N-linked protein glycosylation, z.e., biosynthesis of dolichol pyrophosphate linked oligosaccharide (DLO), which occur in the ER, or transfer of the DLO to asparagine residues of nascent polypeptides. CDG type II involves defects in furtherf processing (synthetic or hydrolytic) of the protein-bound glycan. Currently, twenty -two CDG type I and fourteen type II variants have been identified. One of the most common subtypes of CDG is CDG-Ia (approximately 70% of all CDG cases), which is characterized by loss or reduction of phosphomannomutase 2 (PMM) activity leading to the deficiency or insufficiency in intracellular N-glycosylation (Jaeken et al. J. of Inherit. Met. Disease. 2008, 31 : 669-672). PMM is responsible for the conversion of mannose-6- phosphate to mannose- 1 -phosphate.

[0006] Although several different approaches of developing therapies for CDG have been explored, researchers continue their search for a suitable cure or a therapy for mitigating the disease itself. Existing treatments for manifestations include, for example, nutritional supplements, tube feeding, and a wide range of therapies that attempt to treat gastroesophageal reflux, persistent vomiting, developmental delays, ocular abnormalities, and hypothyroidism. Patients also require intravenous (IV) hydration and physical therapy for stroke-like episodes. Adults with orthopedic symptoms often require wheel chairs, transfer devices, and surgical treatment for scoliosis (Sparks et al., Disorders of Glycosylation Overview. 2005 In: Pagon RA, Adam MP, Bird TD, et al., editors. GeneReviews™. Seattle (WA): University of Washington, Seattle; 1993-2013).

[0007] Currently, CDG-Ib is one known CDG for which a treatment is available, namely oral D-mannose administration. However, such therapy may not be as effective in treating CDG-Ia patients and there are currently limited treatment options for other CDG type I subtypes and CDG type II diseases. One of the reasons for the lack in established therapy for CDG-I disorders may be due to the plethora of heterogeneous clinical phenotypes presented that do not show a direct correlation to the PMM enzyme activity.2 sf-6569471PCT / US25 / 17678 27 February 2025 (27.02.2025)Attorney Docket No.: 74220-20015.40

[0008] Patients suffering from a reduction in PMM activity have reduced productions of mannose- 1 -phosphate (M1P, which is also referred to in the art as Man-l-P), associated with symptoms of multivisceral impairments. In order to overcome PMM production deficiency, it is important to supply downstream enzymes with the required substrate, such as M1P. However, the delivery and maintenance of such a systemic supply of M1P is problematic, as extracellular enzymes within bodily fluids degrade M1P when delivered exogenously by oral or intravenous administration.

[0009] Derivatives of the polar M1P can be synthesized to make M1P more cell- permeable (US Patent Publication No. 2009 / 0054353). This approach, however, is also problematic, as the cell-permeable M1P derivatives have been shown to be either unstable for clinical use or cytotoxic via the by-products of the M1P derivatives (Eklund et al., Glycobiology 2005, 15: 1084-1093; Rutschow et al. Bioorg Med Chem 2002, 10: 4043-4049; and Hardre et al., Bioorg Med Chem Lett 2007, 17: 152-155).

[0010] Other potential therapies have focused on inhibiting enzymes that catabolize mannose-6-phosphate (M6P, also referred to in the art as Man-6-P), a precursor to M1P, via the inhibition of phosphomannose isomerases (PMI). The approach focuses on forcing the reaction towards optimizing homeostasis, which with the use of PMI inhibitors, would have been skewed toward production of M6P. These approaches, however, are ineffective as clinical treatment options due to their associated toxicity, off-target side effects, and poor selective tissue penetration.

[0011] Another potential solution is to use a delivery vehicle (e.g., lipid particles) to encapsulate and deliver M1P (see WO 2015 / 053910). However, due to the high charge and polarity of phosphorylated carbohydrates in general, the optimal delivery vehicle must overcome challenges of stability, phosphorylated carbohydrates loading rate and concentration, toxicity, and delivery efficiency.

[0012] Ataxia is a key driver of disease burden in PMM2-CDG. This condition affects over 95% of PMM2 patients from infancy through adulthood, causing impairment of ambulation, balance, speech, swallowing (choking), and vision. See Pettinato et al., 2021. Ataxia describes poor muscle control that causes clumsy movements. Ataxia can affect walking and balance, hand coordination, speech and swallowing, and eye movements. Ataxia usually results from damage to the part of the brain called the cerebellum or its connections.3 sf-6569471PCT / US25 / 17678 27 February 2025 (27.02.2025)Attorney Docket No.: 74220-20015.40

[0013] Ataxia is commonly quantified by the International Cooperative Ataxia Rating Scale (ICARS), for ages 4 and up. ICARS evaluates: (1) Limb Coordination: Tremor and ataxia in upper & lower body; (2) Postural / Gait: Disturbances in posture and gait; (3) Speech: Fluency & clarity of speech; and (4) Oculomotor: Abnormal eye movements & eye tracking.

[0014] Accordingly, unmet needs exist for improved compositions and methods for delivering phosphorylated carbohydrates, such as M1P and M6P, to treat disorders, such as a congenital disorder of glycosylation (CDG) and ataxia, to subjects (including, for example, humans) in need of such treatment.BRIEF SUMMARY

[0015] In some aspects, provided is a method of treating a congenital disorder of glycosylation in a subject in need thereof, comprising administering to the subject a composition comprising mannose- 1 -phosphate (M1P). In some variations, the M1P is encapsulated in a lipid particle. Lipid particles include, but are not limited to liposomes, micelles, solid lipid nanoparticles, niosomes, lipospheres, emulsomes and emulsions. In some embodiments, the composition is a liposomal composition. In some embodiments, the composition administered may be any of the liposomal compositions described herein. In other variations, the M1P is delivered in a nanoparticle formulation formed from one or more copolymers. For instance, poly (D,L-lactide-co-glycolide) PLGA nanoparticles can be used to deliver the M1P.

[0016] In some embodiments, the congenital disorder of glycosylation is CDG-Ia, CDG- le, CDG-Ii, CDG-Ik, CDG-Io, CDG-Ip, or DPM2-CDG. In one embodiment, the congenital disorder of glycosylation is CDG-Ia (also known as PMM2-CDG). In one embodiment, the disclosure provides a method of treating PMM2-CDG in a patient in need thereof, comprising administering to the patient weekly, or biweekly, or every two weeks an aqueous parenteral (e.g., intravenous) dose of a composition (e.g., a liposomal composition) comprising M1P or a pharmaceutically salt thereof, wherein the dose provides a steady state plasma AUC(o-«>) of M1P of from about 10,000 pg.hr / mL to about 35,000 pg.hr / mL. In some embodiments, administration of a composition (e.g., a liposomal composition) comprising M1P or a pharmaceutically acceptable salt thereof provides a steady state plasma AUC(o-«>) of M1P of from about 12,000 pg.hr / mL to about 30,000 pg.hr / mL. In some embodiments, administration of a composition (e.g., a liposomal composition) comprising M1P or a4 sf-6569471PCT / US25 / 17678 27 February 2025 (27.02.2025)Attorney Docket No.: 74220-20015.40 pharmaceutically salt thereof provides a steady state plasma AUC(o-«>) of M1P of from about 15,000 pg.hr / mL to about 28,000 pg.hr / mL. In some embodiments, administration of a composition (e.g., a liposomal composition) comprising M1P or a pharmaceutically salt thereof provides a steady state plasma AUC(o-«>) of M1P of from about 20,000 pg.hr / mL to about 30,000 pg.hr / mL. In some embodiments, administration of a composition (e.g., a liposomal composition) comprising M1P or a pharmaceutically salt thereof provides a steady state plasma AUC(o-«>) of M1P of from about 15,000 pg.hr / mL to about 25,000 pg.hr / mL.

[0017] In some aspects, provided is a method of treating ataxia in a human suffering from phosphomannomutase 2-congenital disorder of glycosylation (PMM2-CDG), the method comprising administering to the human a liposomal composition comprising mannose 1- phosphate or a pharmaceutically acceptable salt thereof, at a dose between 10 mg / kg and 30 mg / kg of mannose- 1 -phosphate. In some variations, the method further comprises improving the human’s (i) postural and gait disturbances, (ii) limb ataxia, (iii) dysarthria, and / or (iv) oculomotor disorders. In some variations, the method further comprises periodically measuring an International Cooperative Ataxia Rating Scale (ICARS) score for (i) postural and gait disturbances, (ii) limb ataxia, (iii) dysarthria, and / or (iv) oculomotor disorders during the treatment duration in the human; and assessing the human’s ICARS scores over the treatment duration. In some variations, the liposomal composition is administered to the human for a treatment duration over a plurality of weeks, wherein an International Cooperative Ataxia Rating Scale (ICARS) score is measured periodically for (i) postural and gait disturbances, (ii) limb ataxia, (iii) dysarthria, and / or (iv) oculomotor disorders during the treatment duration, and wherein the human’s (i) postural and gait disturbances, (ii) limb ataxia, (iii) dysarthria, and / or (iv) oculomotor disorders improves over the treatment duration based on the ICARS scores.

[0018] In some variations of the foregoing, the subject is a human. In one variation, the subject is an adult. In some embodiments, the compositions are administered weekly, or biweekly, or every two weeks.

[0019] In some embodiments, the M1P is administered as a pharmaceutically acceptable salt. In particular embodiments, the salt is a di-potassium salt. In some embodiments, the salt is a hydrate or a dihydrate.5 sf-6569471PCT / US25 / 17678 27 February 2025 (27.02.2025)Attorney Docket No.: 74220-20015.40

[0020] In any of the foregoing embodiments, the dose of a composition comprising M1P or a pharmaceutically salt thereof can provide a Cmax of M1P in the plasma of from about 150 pg / mL to about 600 pg / mL. In some embodiments, the dose of a composition comprising M1P or a pharmaceutically acceptable salt thereof can provide a Cmax of M1P in the plasma of from about 250 pg / mL to about 500 pg / mL. In some embodiments, the dose of a composition comprising M1P or a pharmaceutically acceptable salt thereof can provide a Cmax of M1P in the plasma of from about 300 pg / mL to about 475 pg / mL. In some variations of the foregoing embodiments, the dose is administered weekly, or biweekly, or every two weeks to achieve the Cmax effects described.

[0021] In some embodiments, the composition comprising M1P or a pharmaceutically salt thereof can provide a steady state plasma AUC(o-«>) of M1P of from about 10,000 pg.hr / mL to about 35,000 pg.hr / mL and a Cmax of from about 150 pg / mL to about 600 pg / mL. In other embodiments, the composition comprising M1P or a pharmaceutically salt thereof can provide a steady state plasma AUC(o-«>) of M1P of from about 12,000 pg.hr / mL to about 30,000 pg.hr / mL and a Cmax of from about 250 pg / mL to about 500 pg / mL. In other embodiments, the composition comprising M1P or a pharmaceutically salt thereof can provide a steady state plasma AUC(o-«>) of M1P of from about 10,000 pg.hr / mL to about 28,000 pg.hr / mL and a Cmax of from about 200 pg / mL to about 480 pg / mL. In some embodiments, the composition comprising M1P or a pharmaceutically salt thereof can provide a steady state plasma AUC(o-«>) of M1P of from about 15,000 pg.hr / mL to about 35,000 pg.hr / mL and a Cmax of from about 400 pg / mL to about 600 pg / mL. In some variations of the foregoing embodiments, the dose is administered weekly, or biweekly, or every two weeks to achieve the Cmax and AUC effects described.

[0022] In some variations, the compositions are administered at a dose of M1P or a pharmaceutically salt thereof from about 10 mg / kg to about 40 mg / kg, from about 20 mg / kg to about 40 mg / kg, from about 30 mg / kg to about 30 mg / kg, from about 25 mg / kg to about 30 mg / kg. In some variations of the foregoing, the dose is administered weekly, or biweekly, or every two weeks at the doses (mg / kg) described. In some variations, the compositions are administered parenterally (e.g., intravenously). For instance, the compositions may be administered by intravenous infusion. It will be understood that in all embodiments, the dose of M1P is based on the molecular weight of the free acid form of M1P (chemical formula C6H13O9P; MW = 260.135 g / mol).6 sf-6569471PCT / US25 / 17678 27 February 2025 (27.02.2025)Attorney Docket No.: 74220-20015.40

[0023] In some embodiments, the compositions are administered as a weekly dose of M1P or a pharmaceutically acceptable salt thereof of about 10 mg / kg, wherein said dose provides a steady state plasma AUC(o-«>) of M1P of from about from about 7,000 pg.hr / mL to about 11,000 pg.hr / mL. In some such embodiments, the dose provides a Cmax of from about 120 pg / mL to about 160 pg / mL.

[0024] In some embodiments, the compositions are administered as a weekly dose of M1P or a pharmaceutically acceptable salt thereof of about 20 mg / kg, wherein said dose provides a steady state plasma AUC(o-«>) of M1P of from about from about 17,000 pg.hr / mL to about 24,000 pg.hr / mL. In some such embodiments, the dose provides a Cmax of from about 260 pg / mL to about 340 pg / mL.

[0025] In some embodiments, the compositions are administered as a weekly dose of M1P or a pharmaceutically acceptable salt thereof of about 30 mg / kg, wherein said dose provides a steady state plasma AUC(o-«>) of M1P of from about from about 28,000 pg.hr / mL to about 36,000 pg.hr / mL. In some such embodiments, the dose provides a Cmax of from about 380 pg / mL to about 400 pg / mL. In other such embodiments, the dose provides a Cmax of from about 400 pg / mL to about 500 pg / mL

[0026] It has been surprisingly discovered that plasma concentrations of M1P greater than 100 pg / mL (e.g., greather than 200 pg / mL, greather than 300 pg / mL, greater than 400 pg / mL or greater than 500 pg / mL) can be achieved following administration of the disclosed compositions, and that these concentrations result in significant cellular uptake of M1P, particularly in liver cells. These concentrations of M1P in the plasma are associated with significant restoration of GDP -Mannose in liver cells. For instance, in particular embodiments, the levels of GDP -Mannose in liver cells can be restored to normal levels observed in healthy humans.

[0027] In some embodiments, the concentration of M1P in the plasma following administration of a composition disclosed herein is from about 10 pg / mL to about 40 pg / mL, wherein the concentration of M1P is measured ar the trough level (i.e., immediately before the next dose is administered). In other embodiments, the concentration of M1P in the plasma following administration of a composition disclosed herein is from about 20 pg / mL to about 40 pg / mL, wherein the concentration of M1P is measured ar the trough level. In other embodiments, the concentration of M1P in the plasma following administration of a7 sf-6569471PCT / US25 / 17678 27 February 2025 (27.02.2025)Attorney Docket No.: 74220-20015.40 composition disclosed herein is from about 20 pg / mL to about 30 pg / mL, wherein the concentration of M1P is measured ar the trough level. In other embodiments, the concentration of M1P in the plasma following administration of a composition disclosed herein is from about 25 pg / mL to about 30 pg / mL, wherein the concentration of M1P is measured ar the trough level.

[0028] In some embodiments, the disclosed composotions of M1P are administered at a dose that achieves a plasma concentraion of greater than 20 pg / mL for at least 20% of time over a dosing cycle (e.g., weekly, or biweekly, or every two weeks). In other embodiments, the disclosed composotions of M1P are administered at a dose that achieves a plasma concentraion of greater than 20 pg / mL for at least 30% of time over a dosing cycle (e.g., weekly, or biweekly, or every two weeks). In other embodiments, the disclosed composotions of M1P are administered at a dose that achieves a plasma concentraion of greater than 20 pg / mL for at least 40% of time over a dosing cycle (e.g., weekly, or biweekly, or every two weeks). In other embodiments, the disclosed composotions of M1P are administered at a dose that achieves a plasma concentraion of greater than 20 pg / mL for at least 50% of time over a dosing cycle (e.g., weekly, or biweekly, or every two weeks). In other embodiments, the disclosed composotions of M1P are administered at a dose that achieves a plasma concentraion of greater than 20 pg / mL for at least 60% of time over a dosing cycle (e.g., weekly, or biweekly, or every two weeks). In other embodiments, the disclosed composotions of M1P are administered at a dose that achieves a plasma concentraion of greater than 20 pg / mL for at least 70% of time over a dosing cycle (e.g., weekly, or biweekly, or every two weeks). In other embodiments, the disclosed composotions of M1P are administered at a dose that achieves a plasma concentraion of greater than 20 pg / mL for at least 80% of time over a dosing cycle (e.g., weekly, or biweekly, or every two weeks). In other embodiments, the disclosed composotions of M1P are administered at a dose that achieves a plasma concentraion of greater than 20 pg / mL for at least 90% of time over a dosing cycle (e.g., weekly, or biweekly, or every two weeks). In other embodiments, the disclosed composotions of M1P are administered at a dose that achieves a plasma concentraion of greater than 20 pg / mL from about 20% to about 90% of time over a dosing cycle (e.g., weekly, or biweekly, or every two weeks). In other embodiments, the disclosed composotions of M1P are administered at a dose that achieves a plasma concentraion of greater than 20 pg / mL from about 30% to about 85% of time over a dosing cycle (e.g., weekly, or biweekly, or every two weeks). In other embodiments, the8 sf-6569471PCT / US25 / 17678 27 February 2025 (27.02.2025)Attorney Docket No.: 74220-20015.40 disclosed composotions of M1P are administered at a dose that achieves a plasma concentraion of greater than 20 pg / mL from about 50% to about 80% of time over a dosing cycle (e.g., weekly, or biweekly, or every two weeks).

[0029] In some embodiments, the disclosed composotions of M1P are administered at a dose that achieves a plasma concentraion of greater than 40 pg / mL for at least 20% of time over a dosing cycle (e.g., weekly, or biweekly, or every two weeks). In other embodiments, the disclosed composotions of M1P are administered at a dose that achieves a plasma concentraion of greater than 40 pg / mL for at least 30% of time over a dosing cycle (e.g., weekly, or biweekly, or every two weeks). In other embodiments, the disclosed composotions of M1P are administered at a dose that achieves a plasma concentraion of greater than 40 pg / mL for at least 40% of time over a dosing cycle (e.g., weekly, or biweekly, or every two weeks). In other embodiments, the disclosed composotions of M1P are administered at a dose that achieves a plasma concentraion of greater than 40 pg / mL for at least 50% of time over a dosing cycle (e.g., weekly, or biweekly, or every two weeks). In other embodiments, the disclosed composotions of M1P are administered at a dose that achieves a plasma concentraion of greater than 40 pg / mL for at least 60% of time over a dosing cycle (e.g., weekly, or biweekly, or every two weeks). In other embodiments, the disclosed composotions of M1P are administered at a dose that achieves a plasma concentraion of greater than 40 pg / mL for at least 70% of time over a dosing cycle (e.g., weekly, or biweekly, or every two weeks). In other embodiments, the disclosed composotions of M1P are administered at a dose that achieves a plasma concentraion of greater than 40 pg / mL for at least 80% of time over a dosing cycle (e.g., weekly, or biweekly, or every two weeks). In other embodiments, the disclosed composotions of M1P are administered at a dose that achieves a plasma concentraion of greater than 40 pg / mL for at least 90% of time over a dosing cycle (e.g., weekly, or biweekly, or every two weeks). In other embodiments, the disclosed composotions of M1P are administered at a dose that achieves a plasma concentraion of greater than 40 pg / mL from about 20% to about 90% of time over a dosing cycle (e.g., weekly, or biweekly, or every two weeks). In other embodiments, the disclosed composotions of M1P are administered at a dose that achieves a plasma concentraion of greater than 40 pg / mL from about 30% to about 85% of time over a dosing cycle (e.g., weekly, or biweekly, or every two weeks). In other embodiments, the disclosed composotions of M1P are administered at a dose that achieves a plasma9 sf-6569471PCT / US25 / 17678 27 February 2025 (27.02.2025)Attorney Docket No.: 74220-20015.40 concentraion of greater than 40 pg / mL from about 50% to about 80% of time over a dosing cycle (e.g., weekly, or biweekly, or every two weeks).

[0030] In any of the foreoing embodiments, the half-life (T1 / 2) of M1P following administration of a composition disclosed herein is from about 50 hous to about 175 hours. In some embodiments, the half-life of M1P is from about 75 hours to about 150 hours. In some embodiments, the half-life of M1P is from about 70 hours to about 120 hours.

[0031] It has been found that the compositions disclosed herein can be administered safely and efficaciously to patients in need thereof. For instance, the compositions can effectively restore the levels of GDP -mannose in the patient by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, or at least 90%. In some embodiments, the compositions restore the levels of GDP -mannose in the patient of from about 20% to about 80%. In other embodiments, the compositions restore the levels of GDP-mannose in the patient of from about 30% to about 70%. In other embodiments, the compositions restore the levels of GDP-mannose in the patient of from about 40% to about 60%.

[0032] In some embodiments, the liposomal comprise comprising three or more lipids. In some embodiments, the MIPlipid ratio is from about 0.1 to about 2, from about 0.1 to about 1, from about 0.1 to about 0.5, from about 0.1 to about 0.4, from about 0.1 to about 0.3, from about 0.1 to about 0.2, or from about 0.5 to about 1.5. In some variations, the MIPlipid ratio is about 0.1, 0.2, or 0.3. In some embodiments, the liposomal compositions comprise a phosphoethanolamine lipid. In other embodiments, the liposomal compositions comprise a phosphocholine lipid. In other embodiments, the liposomal compositions comprise a phosphoethanolamine lipid and a phosphocholine lipid.

[0033] In some embodiments, the liposomal compositions include a lipid membrane enclosing an intraliposomal compartment, in which the liposomes encapsulate in the intraliposomal compartment mannose- 1 -phosphate (M1P). The composition further includes intraliposomal buffer comprising a buffer salt, and optionally acid. In some variations, the pKa of the buffer salt in the intraliposomal buffer is between 6 to 8.5. The composition further includes extraliposomal buffer comprising a buffer salt and a tonicity modifier. In some variations, the pKa of the buffer salt of the extraliposomal buffer is between 6.0 to 8.5.10 sf-6569471PCT / US25 / 17678 27 February 2025 (27.02.2025)Attorney Docket No.: 74220-20015.40In some variation, the compositions optionally further include a radical scavenging antioxidant, present in the lipid bilayer.

[0034] In some embodiments, the lipid membrane comprises: (a) at least one phospholipid having an ethanolamine head group and at least one unsaturated fatty acid tail; (b) at least one phospholipid having a choline group and at least one unsaturated fatty acid tail; and (c) at least one phospholipid having an ethanolamine head group and at least one saturated fatty acid tail, conjugated to polyethylene glycol (PEG). In certain variations, the lipid membrane comprises DOPE, DOPC and DSPE conjugated to PEG.

[0035] In some embodiments, the intraliposomal buffer comprises tris(hydroxymethyl)aminomethane (Tris). In some embodiments, the extraliposomal buffer comprises Tris and saline. In other embodiments, the extraliposomal buffer comprises Tris and sugars, such as sucrose. In some embodiments, the radical scavenging antioxidant comprises butylated hydroxytoluene (BHT). In some embodiments, the radical scavenging antioxidant is absent.

[0036] In one variation, the liposomal compositions comprise: liposomes having a lipid membrane enclosing an intraliposomal compartment, wherein the liposomes encapsulate in the intraliposomal compartment M1P, and wherein the lipid membrane comprises DOPC, DOPE, and DSPE conjugated to PEG; intraliposomal buffer comprising Tris; extraliposomal buffer comprising Tris and saline; and BHT.

[0037] In one variation, the liposomal compositions comprise: liposomes having a lipid membrane enclosing an intraliposomal compartment, wherein the liposomes encapsulate in the intraliposomal compartment M1P, and wherein the lipid membrane comprises DOPC, DOPE, and DSPE conjugated to PEG; intraliposomal buffer comprising Tris; and extraliposomal buffer comprising Tris and saline.BRIEF DESCRIPTION OF DRAWINGS

[0038] The patent or application file contains at least one drawing executed in color. Copies of this patent or patent application publication with color drawings will be provided by the Office upon request and payment of the necessary fee.11 sf-6569471PCT / US25 / 17678 27 February 2025 (27.02.2025)Attorney Docket No.: 74220-20015.40

[0039] The present application can be understood by reference to the following description taken in conjunction with the accompanying figures.

[0040] FIG.l shows plasma concentration of M1P following three different liposomal compositions comprising various doses of M1P (3 mg / kg, 10 mg / kg and 20 mg / kg) after the third dose of a weekly dosing schedule.

[0041] FIGS. 2A-2C show that Formulation A is an effective MIP-liposome formulation in PMM2-CDG patient-derived fibroblasts. FIG. 2A shows GDP -mannose levels in healthy fibroblasts (Normal) and GM20942 (PMM2) fibroblasts derived from patients with phosphomannomutase 2 congenital disorder of glycosylation (PMM2-CDG). The * indicates significant (P<0.05) difference between groups as determined by t-test. FIG. 2B shows Formulation A increases GDP-mannose levels in fibroblasts from PMM2-CDG cells from patients with different genotypes. Total GDP-mannose was quantified in cells treated for 24 hours with 0.5 mM Formulation A or vehicle (untreated). FIG. 2C shows Con A western blot analysis of total proteins from Formulation A-treated or untreated GM20942 cells. Untreated healthy fibroblasts (HDF) were used as control. Triplicate samples were run for each condition. Protein blots were probed with either Concanavalin A (Con A) to detect glycoproteins or anti -glyceraldehyde 3-phosphate dehydrogenase (GAPDH) antibody. Molecular weight markers are shown in FIG. 2D, which provide western blot of total proteins from Formulation A-treated (increasing concentrations) GM20942 cells or untreated HDF. Blot was probed with either an anti-intercellular adhesion molecule- 1 (ICAM-1) or anti-GAPDH antibodies. Numbers at the left indicate molecular weight markers.

[0042] FIG. 3 shows N-gly comics profile of healthy control cell lines. Cells were processed for N-glycan analysis as described in Materials and Methods. For each N-glycan( with predicted structure) detected, the relative abundance versus total N-glycan content in the sample was calculated and reported as a percentage. For maximum clarity, N-glycan structures were divided into non-fucosylated asialo glycans (red), non-fucosylated glycans (blue), high mannose glycans (black) and sialylated glycans (purple).

[0043] FIG. 4 shows N-glycomics profile of CDG patient-derived cell lines. Dermal fibroblast or LCL cell lines derived from PMM2-, DPMI-, ALG1-, ALG3-, ALG6-, ALG12- or DPAGT1-CDG patients were characterized (N=3 for each cell type and treatment). For each N-glycan structure detected, the relative abundance versus total N-glycan content in the12 sf-6569471PCT / US25 / 17678 27 February 2025 (27.02.2025)Attorney Docket No.: 74220-20015.40 sample was calculated and reported as a percentage. The fold change in relative abundance in CDG versus healthy control cells was calculated for each glycan. The mean fold change for triplicate samples was calculated and represented as a heat map. N-glycans structures are shown on the left and are grouped from top to bottom into fucosylated asialo (red), non- fucosylated asialo (blue), high mannose (black) and sialylated (purple). Separate fold change scales are shown for fibroblasts (left) and LCL (right) lines.

[0044] FIG. 5 shows Formulation A restores the high-mannose N-glycan profile of PMM2-CDG patient derived fibroblasts. PMM2-CDG fibroblasts were treated with vehicle (N=3) or ImM Formulation A (N=3) for 24 hrs and total cellular N-glycan profile was characterized. Untreated HDF (N=3) were used as the healthy control. The percent of total N- glycans was calculated and reported for each high mannose glycan. One-way anova was used to assess significant differences between treatment groups. Significance is indicated with * (p<0.05), ** (p<0.01), or *** (p<0.001).

[0045] FIG. 6 shows Formulation A treatment effect on CDG patient-derived cells in vitro. PMM2-, DPMI-, ALG1-, ALG3-, ALG6-, ALG12- or DPAGT1-CDG cells were treated with either 1 mM of Formulation A or vehicle control (untreated) for 24 hours (N=3 for each cell type and treatment). Untreated HDF or parental control LCL were used as the healthy control (N=3). For each N-glycan structure detected, the relative abundance versus total N-glycan content in the sample was calculated. The fold change in N-glycan abundance in Formulation A-treated versus vehicle -treated cells was calculated. The mean fold change for triplicate samples from each cell type was caclucated and represented as a heat map. N- glycans structures are shown on the left and are grouped from top to bottom into fucosylated asialo (red), non-fucosylated asialo (blue), high mannose (black) and sialylated (purple). The scale for the fold change between Formulation A vs. vehicle (untreated) cells is shown on the right.

[0046] FIGS. 7A-7C show near infra-red fluorescence (NIRF) imaging of liposome- encapsulated sulfo-Cy5.5 (Cy5.5). Ventral whole body imaging of vehicle control (N=l), free Cy5.5 (N=3) or Cy5.5-liposome (N=3) treated BALB / c mice was performed at pre-dose and 4 hours after treatment. FIG. 7A shows quantification of fluorescence from ex vivo imaging of different tissues at 4 hours showed the strongest fluorescence in liver and spleen, compared to other tissues. Mean fluorescence shown as photons / s / cmA2 / sr. FIG. 7B shows13 sf-6569471PCT / US25 / 17678 27 February 2025 (27.02.2025)Attorney Docket No.: 74220-20015.40 comparison of ex vivo imaging in different tissues at 4 hours (lymph node (LN), heart, lung, brain, kidney, liver, spleen, muscle, small intestine (SI), and colon (C).

[0047] FIGS. 8A-8C shows in vivo pharmacokinetic characterization of M1P liposomes. Pharmacokinetic profile of Formulation A (10 mg / kg) in rats by measuring mannose-1- phosphate (M1P) showing Formulation A has a half-life of 16.2 hours (FIG. 8A).Pharmacokinetic profile of Formulation A (20 mg / kg) by measuring M1P in mouse, showing Formulation A has a half-life of 11.2 hours (FIG. 8B), and in dogs (FIG. 8C) by measuring M1P at different dose levels ( 4.8, 25.2 and 46.8 mg / kg, with half-lives of 8.2, 24.8 and 40.2 hours, respectively).

[0048] FIG. 9A depicts a graph showing the ICARS Score with Formulation A. FIG. 9B and FIG. 9C depict graphs showing the change in ICARS Score with Formulation A.

[0049] FIGS. 10A-10B depict the Archimedes Spiral (component of ICARS) in Subject 2007. FIG. 10A shows the baseline Archimedes Spiral and FIG. 10B shows the Archimedes Spiral at Week 12. FIGS. 10C-10D depict the Archimedes Spiral (component of ICARS) in Subject 2008. FIG. 10C shows the baseline Archimedes Spiral and FIG. 10D shows the Archimedes Spiral at Week 12.

[0050] FIG. 11 depicts a graph showing the change in ICARS Score in a comparison across studies in PMM2.DETAILED DESCRIPTION

[0051] The following description sets forth exemplary methods, compositions, parameters, and the like. It should be recognized, however, that such description is not intended as a limitation on the scope of the present disclosure but is instead provided as a description of exemplary embodiments.

[0052] Provided herein are methods of treating a congenital disorder of glycosylation (CDG) in a subject in need thereof by administering liposomal compositions comprising mannose- 1 -phosphate (M1P). Variations of the liposomal compositions that may be used are described herein. Congenital disorders of glycosylation include, for example, CDG-Ia, CDG- le, CDG-Ii, CDG-Ik, CDG-Io, CDG-Ip, or DPM2-CDG. In one embodiment, the methods herein treat phosphomannomutase 2-congenital disorder of glycosylation (PMM2-CDG), formerly known as congenital disorder of glycosylation (CDG) type la.14 sf-6569471PCT / US25 / 17678 27 February 2025 (27.02.2025)Attorney Docket No.: 74220-20015.40

[0053] In some embodiments, compositions comprising M1P encapsulated by liposomes are used in the methods herein.

[0054] Provided herein are methods of treating ataxia in a human suffering from phosphomannomutase 2-congenital disorder of glycosylation (PMM2-CDG). The method comprises: administering to the human a liposomal composition comprising mannose 1- phosphate or a pharmaceutically acceptable salt thereof. The methods provided herein improve the human’s (i) postural and gait disturbances, (ii) limb ataxia, (iii) dysarthria, and / or (iv) oculomotor disorders. In some variations, the method further comprises: periodically measuring an International Cooperative Ataxia Rating Scale (ICARS) score for (i) postural and gait disturbances, (ii) limb ataxia, (iii) dysarthria, and / or (iv) oculomotor disorders during the treatment duration in the human; and assessing the human’s ICARS scores over the treatment duration.

[0055] In some embodiments, the liposomal composition is administered to the human for a treatment duration of at least 2 weeks, at least 3 weeks, at least 4 weeks, or at least 6 months, or at least 1 year. In some variations, when an International Cooperative Ataxia Rating Scale (ICARS) score is measured periodically over the treatment duration for (i) postural and gait disturbances, (ii) limb ataxia, (iii) dysarthria, and / or (iv) oculomotor disorders during the treatment duration, the human’s (i) postural and gait disturbances, (ii) limb ataxia, (iii) dysarthria, and / or (iv) oculomotor disorders improves over the treatment duration based on the ICARS scores.

[0056] Unless defined otherwise, all scientific and technical terms are understood to have the same meaning as commonly used in the art to which they pertain. For the purpose of the present disclosure, the following terms are defined.

[0057] The term “about” as used herein refers to the usual error range for the respective value readily known to the skilled person in this technical field. Reference to “about” a value or parameter herein includes (and describes) embodiments that are directed to that value or parameter per se. For example, “about x” includes and describes “x” per se. In some embodiments, the term “about” when used in association with a measurement, or used to modify a value, a unit, a constant, or a range of values, refers to variations of + / - 2%.

[0058] Reference to “between” two values or parameters herein includes (and describes) embodiments that include those two values or parameters per se. For example, description15 sf-6569471PCT / US25 / 17678 27 February 2025 (27.02.2025)Attorney Docket No.: 74220-20015.40 referring to “between x and y” includes description of “x” and “y” per se. Further, it should also be understood that “between x and y” can also be expressed as “about x to y” or “about x-y”.

[0059] As used herein, the singular forms “a,” “an,” and “the” include plural reference unless the context clearly indicates otherwise.Methods of Treatment

[0060] In some aspects, provided is a method for treating a congenital disorder of glycosylation (CDG) in a subject in need thereof, comprising administering to the subject a liposomal composition comprising mannose- 1 -phosphate (M1P). In one variation, the methods herein treat phosphomannomutase 2-congenital disorder of glycosylation (PMM2- CDG), formerly known as congenital disorder of glycosylation (CDG) type la.

[0061] In some aspects, provided is a method of treating ataxia in a human suffering from phosphomannomutase 2-congenital disorder of glycosylation (PMM2-CDG), the method comprising administering to the human a liposomal composition comprising mannose 1- phosphate or a pharmaceutically acceptable salt thereof, at a dose between 10 mg / kg and 30 mg / kg of mannose- 1 -phosphate. In some variations, the method further comprises improving the human’s (i) postural and gait disturbances, (ii) limb ataxia, (iii) dysarthria, and / or (iv) oculomotor disorders. In some variations, the method further comprises periodically measuring an International Cooperative Ataxia Rating Scale (ICARS) score for (i) postural and gait disturbances, (ii) limb ataxia, (iii) dysarthria, and / or (iv) oculomotor disorders during the treatment duration in the human; and assessing the human’s ICARS scores over the treatment duration. In some variations, the liposomal composition is administered to the human for a treatment duration over a plurality of weeks, wherein an International Cooperative Ataxia Rating Scale (ICARS) score is measured periodically for (i) postural and gait disturbances, (ii) limb ataxia, (iii) dysarthria, and / or (iv) oculomotor disorders during the treatment duration, and wherein the human’s (i) postural and gait disturbances, (ii) limb ataxia, (iii) dysarthria, and / or (iv) oculomotor disorders improves over the treatment duration based on the ICARS scores.

[0062] The liposomal compositions described herein are useful for delivering M1P to a subject in need thereof.16 sf-6569471PCT / US25 / 17678 27 February 2025 (27.02.2025)Attorney Docket No.: 74220-20015.40

[0063] In some embodiments, the subject is a mammal, such as a human, domestic animal, such as a feline or canine subject, farm animal (e.g., bovine, equine, caprine, ovine, and porcine subject), wild animal (whether in the wild or in a zoological garden), research animal, such as mouse, rat, rabbit, goat, sheep, pig, dog, and cat, and birds. In one embodiment, the subject is a human. In one variation, the subject is an adult human.

[0064] In some variations, the subject may be at risk. For example, in one variation, the subject at risk is a human. A subject at risk of developing a particular disease, disorder, or condition, such as a congenital disorder of glycosylation, may or may not have detectable disease or symptoms of disease, and may or may not have displayed detectable disease or symptoms of disease prior to the treatment methods described herein. In certain variations, an individual “at risk” is an individual having risk factors, which are measurable parameters that correlate with development of a particular disease, disorder, or condition, as known in the art. A subject having one or more of these risk factors has a higher probability of developing a particular disease, disorder, or condition such as a congenital disorder of glycosylation, than a subject without one or more of these risk factors.

[0065] In some embodiments, congenital disorders of glycosylation (CDG) is a group of genetic disorders that result in errors of metabolism in which glycosylation of a variety of tissue proteins and / or lipids is deficient or defective. Congenital disorders of glycosylation may also be known as CDG syndromes. CDG syndromes may often cause serious, occasionally fatal, malfunction of several different organ systems, such as the nervous system, brain, muscles, and intestines, in affected infants. Manifestations of CDG syndromes may range from severe developmental delay and hypotonia beginning in infancy, to hypoglycemia and protein-losing enteropathy with normal development. Developmental delay can be a common initial indication for a CDG diagnosis. One of the most common subtype of CDG syndromes is CDG-Ia (also known as PMM2-CDG) where the genetic defect leads to the loss of phosphomannomutase 2, which is the enzyme responsible for the conversion of mannose-6-phosphate into mannose- 1 -phosphate.

[0066] CDG syndromes may be classified as type I (CDG-I) and type II (CDG-II). Such classification may depend on the nature and location of the biochemical defect in the metabolic pathway relative to the action of oligosaccharyltransferase. Methods for screening for CDG subtype may include the analysis of transferrin glycosylation status by, for example, isoelectric focusing or ESI-MS. CDG type I include, for example, la (PMM2-CDG), lb (MPI-17 sf-6569471PCT / US25 / 17678 27 February 2025 (27.02.2025)Attorney Docket No.: 74220-20015.40CDG), Ic (ALG6-CDG) , Id (ALG3-CDG), le (DPM1-CDG), If (MPDU1-CDG), Ig (ALG12-CDG), Ih (ALG8-CDG), li (ALG2-CDG), Ij (DPAGT1-CDG), Ik (ALG1-CDG), IL (ALG9-CDG), Im (DOLK-CDG), In (RFT1-CDG), Io (DPM3-CDG), Ip (ALG11-CDG), Iq (SRD5A3-CDG), Ir (DDOST-CDG), DPM2-CDG, TUSC3-CDG, MAGT1-CDG, DHDDS- CDG, and I / IIx. CDG type II include, for example, Ila (MGAT2-CDG), lib (GCS1-CDG), lie (SLC335C1-CDG), lid (B4GALT1-CDG), lie (COG7-CDG), Ilf (SLC35A1-CDG), Ilg (COG1-CDG), Ilh (COG8-CDG), Hi (COG5-CDG), Ilj (COG4-CDG), IIL (COG6-CDG), ATP6V0A2-CDG, MAN1B1-CDG, and ST3GAL3-CDG.

[0067] Congenital disorders of glycosylation (CDG) that may be treated with the liposomal compositions provided herein include, for example, la (PMM2-CDG), lb (MPI- CDG), Ic (ALG6-CDG) , Id (ALG3-CDG), le (DPMI -CDG), If (MPDU1-CDG), Ig (ALG12-CDG), Ih (ALG8-CDG), li (ALG2-CDG), Ij (DPAGT1-CDG), Ik (ALG1-CDG), IL (ALG9-CDG), Im (DOLK-CDG), In (RFT1-CDG), Io (DPM3-CDG), Ip (ALG11-CDG), Iq (SRD5A3-CDG), Ir (DDOST-CDG), DPM2-CDG, TUSC3-CDG, MAGT1-CDG, DHDDS- CDG, I / IIx, Ila (MGAT2-CDG), lib (GCS1-CDG), lie (SLC335C1-CDG), lid (B4GALT1- CDG), lie (COG7-CDG), Ilf (SLC35A1-CDG), Ilg (COG1-CDG), Ilh (COG8-CDG), Hi (COG5-CDG), Ilj (COG4-CDG), IIL (COG6-CDG), ATP6V0A2-CDG, MAN1B1-CDG, and ST3GAL3-CDG. In some variations, the compositions and methods described herein are suitable to treat CDG-Ia, CDG-Ib, CDG-Ie, CDG-Ig, CDG-Ii, CDG-Ik, CDG-Io, CDG-Ip, or DPM2-CDG.

[0068] In some embodiments, “treatment” or “treating” includes an approach for obtaining beneficial or desired results including clinical results. Beneficial or desired clinical results may include one or more of the following: a) inhibiting the disease or condition (e.g., decreasing one or more symptoms resulting from the disease or condition, and / or diminishing the extent of the disease or condition); b) slowing or arresting the development of one or more clinical symptoms associated with the disease or condition (e.g., stabilizing the disease or condition, preventing or delaying the worsening or progression of the disease or condition, and / or preventing or delaying the spread of the disease or condition); and / or c) relieving the disease, that is, causing the regression of clinical symptoms (e.g., ameliorating the disease state, providing partial or total remission of the disease or condition, enhancing effect of another medication, delaying the progression of the disease, increasing the quality of life, and / or prolonging survival.18 sf-6569471PCT / US25 / 17678 27 February 2025 (27.02.2025)Attorney Docket No.: 74220-20015.40

[0069] In some embodiments, “prevention” or “preventing” includes any treatment of a disease or condition that causes the clinical symptoms of the disease or condition not to develop. Compounds may, in some embodiments, be administered to a subject (including a human) who is at risk or has a family history of the disease or condition.

[0070] In some variations, an “effective amount” is at least an amount effective, at dosages and for periods of time necessary, to achieve the desired therapeutic or prophylactic result. An effective amount can be provided in one or more administrations.

[0071] In some variations, a “therapeutically effective amount” is at least the minimum concentration required to effect a measurable improvement of a particular disease, disorder, or condition, such as a congenital disorder of glycosylation. A therapeutically effective amount herein may vary according to factors such as the disease state, genotype, age, sex, and weight of the subject, and the ability of the lipid compositions of the present disclosure to elicit a desired response in the subject. A therapeutically effective amount is also one in which any toxic or detrimental effects of the lipid compositions of the present disclosure are outweighed by the therapeutically beneficial effects.

[0072] In some variations, the compositions are administered at a dose of M1P or a pharmaceutically salt thereof from about 10 mg / kg to about 40 mg / kg, from about 20 mg / kg to about 40 mg / kg, from about 30 mg / kg to about 30 mg / kg, from about 25 mg / kg to about 30 mg / kg. In some variations, the dose is administered weekly, or biweekly, or every two weeks. In some variations, the compositions are administered parenterally (e.g., intravenously). For instance, the compositions may be administered by intravenous infusion. It will be understood that in all embodiments, the dose of M1P is based on the molecular weight of the free acid form of M1P (chemical formula C6H13O9P; MW = 260.135 g / mol).

[0073] In some embodiments, the subject has a total ICARS score of greater than about 20. In some embodiments, the subject has a total ICARS score of less than about 80. In some embodiments, the subject has a total ICARS score of between about 20 and about 80. In some embodiments, the subject has a total ICARS score of greater than 20. In some embodiments, the subject has a total ICARS score of less than 80. In some embodiments, the subject has a total ICARS score of between 20 and 80. In some embodiments, the subject has a total ICARS score of > 20 and < 80.19 sf-6569471PCT / US25 / 17678 27 February 2025 (27.02.2025)Attorney Docket No.: 74220-20015.40

[0074] In one embodiment, the disclosure provides a method of treating PMM2-CDG in a patient in need thereof, comprising administering to the patient weekly or biweekly or every 2 weeks an aqueous parenteral (e.g., intravenous) dose of a composition (e.g., a liposomal composition) comprising M1P or a pharmaceutically salt thereof, wherein the dose provides a steady state plasma AUC(o-«>) of M1P of from about 10,000 pg.hr / mL to about 35,000 pg.hr / mL. In some variation sof the foregoing embodiment, the dose is administered weekly, or biweekly, or every two weeks to the patient to achieve the AUC effects described. It will be understood that reference to steady state plasma AUC(o-«>) or Cmax of M1P refers to the total amount of M1P present in the plasma, which constitutes M1P encapsulated in the liposome and free M1P not associated with the liposome. In some embodiments, administration of a composition (e.g., a liposomal composition) comprising M1P or a pharmaceutically acceptable salt thereof provides a steady state plasma AUC(o-«>) of M1P of from about 12,000 pg.hr / mL to about 30,000 pg.hr / mL. In some embodiments, administration of a composition (e.g., a liposomal composition) comprising M1P or a pharmaceutically salt thereof provides a steady state plasma AUC(o-«>) of M1P of from about 15,000 pg.hr / mL to about 28,000 pg.hr / mL. In some embodiments, administration of a composition (e.g., a liposomal composition) comprising M1P or a pharmaceutically salt thereof provides a steady state plasma AUC(o-«>) of M1P of from about 20,000 pg.hr / mL to about 30,000 pg.hr / mL. In some embodiments, administration of a composition (e.g., a liposomal composition) comprising M1P or a pharmaceutically salt thereof provides a steady state plasma AUC(o-«>) of M1P of from about 15,000 pg.hr / mL to about 25,000 pg.hr / mL.

[0075] In some embodiments, the M1P is administered as a pharmaceutically acceptable salt. In particular embodiments, the salt is a di-potassium salt. In some embodiments, the salt is a hydrate or a dihydrate.

[0076] In any of the foregoing embodiments, the weekly or biweekly or every 2 weeks dose of a composition comprising M1P or a pharmaceutically salt thereof can provide a Cmax of M1P in the plasma of from about 150 pg / mL to about 600 pg / mL. In some embodiments, the weekly or biweekly or every 2 weeks dose of a composition comprising M1P or a pharmaceutically acceptable salt thereof can provide a Cmax of M1P in the plasma of from about 250 pg / mL to about 500 pg / mL. In some embodiments, the weekly or biweekly or every 2 weeks dose of a composition comprising M1P or a pharmaceutically20 sf-6569471PCT / US25 / 17678 27 February 2025 (27.02.2025)Attorney Docket No.: 74220-20015.40 acceptable salt thereof can provide a Cmax of M1P in the plasma of from about 300 pg / mL to about 475 pg / mL.

[0077] In some embodiments, the weekly or biweekly or every 2 weeks dose of a composition comprising M1P or a pharmaceutically salt thereof can provide a steady state plasma AUC(o-«>) of M1P of from about 10,000 pg.hr / mL to about 35,000 pg.hr / mL and a Cmax of from about 150 pg / mL to about 600 pg / mL. In other embodiments, the weekly or biweekly or every 2 weeks dose of a composition comprising M1P or a pharmaceutically salt thereof can provide a steady state plasma AUC(o-«>) of M1P of from about 12,000 pg.hr / mL to about 30,000 pg.hr / mL and a Cmax of from about 250 pg / mL to about 500 pg / mL. In other embodiments, the weekly or biweekly or every 2 weeks dose of a composition comprising M1P or a pharmaceutically salt thereof can provide a steady state plasma AUC(o-«>) of M1P of from about 10,000 pg.hr / mL to about 28,000 pg.hr / mL and a Cmax of from about 200 pg / mL to about 480 pg / mL. In some embodiments, the weekly or biweekly or every 2 weeks dose of a composition comprising M1P or a pharmaceutically salt thereof can provide a steady state plasma AUC(o-«>) of M1P of from about 15,000 pg.hr / mL to about 35,000 pg.hr / mL and a Cmax of from about 400 pg / mL to about 600 pg / mL.

[0078] In some embodiments, the compositions are administered as a weekly dose of M1P or a pharmaceutically acceptable salt thereof of about 10 mg / kg, wherein said dose provides a steady state plasma AUC(o-«>) of M1P of from about from about 7,000 pg.hr / mL to about 11,000 pg.hr / mL. In some such embodiments, the dose provides a Cmax of from about 120 pg / mL to about 160 pg / mL.

[0079] In some embodiments, the compositions are administered as a weekly dose of M1P or a pharmaceutically acceptable salt thereof of about 20 mg / kg, wherein said dose provides a steady state plasma AUC(o-«>) of M1P of from about from about 17,000 pg.hr / mL to about 24,000 pg.hr / mL. In some such embodiments, the dose provides a Cmax of from about 260 pg / mL to about 340 pg / mL.

[0080] In some embodiments, the compositions are administered as a weekly dose of M1P or a pharmaceutically acceptable salt thereof of about 30 mg / kg, wherein said dose provides a steady state plasma AUC(o-«>) of M1P of from about from about 28,000 pg.hr / mL to about 36,000 pg.hr / mL. In some such embodiments, the dose provides a Cmax of from21 sf-6569471PCT / US25 / 17678 27 February 2025 (27.02.2025)Attorney Docket No.: 74220-20015.40 about 380 pg / mL to about 400 pg / mL. In other such embodiments, the dose provides a Cmax of from about 400 pg / mL to about 500 pg / mL

[0081] It has been surprisingly discovered that plasma concentrations of M1P greater than 100 pg / mL (e.g., greather than 200 pg / mL, greather than 300 pg / mL, greater than 400 pg / mL or greater than 500 pg / mL) can be achieved following administration of the disclosed compositions, and that these concentrations result in significant cellular uptake of M1P, particularly in liver cells. These concentrations of M1P in the plasma are associated with significant restoration of GDP -Mannose in liver cells. For instance, in particular embodiments, the levels of GDP -Mannose in liver cells can be restored to normal levels observed in healthy humans.

[0082] In some embodiments, the concentration of M1P in the plasma following administration of a composition disclosed herein is from about 10 pg / mL to about 40 pg / mL, wherein the concentration of M1P is measured ar the trough level (i.e., immediately before the next dose is administered). In other embodiments, the concentration of M1P in the plasma following administration of a composition disclosed herein is from about 20 pg / mL to about 40 pg / mL, wherein the concentration of M1P is measured ar the trough level. In other embodiments, the concentration of M1P in the plasma following administration of a composition disclosed herein is from about 20 pg / mL to about 30 pg / mL, wherein the concentration of M1P is measured ar the trough level. In other embodiments, the concentration of M1P in the plasma following administration of a composition disclosed herein is from about 25 pg / mL to about 30 pg / mL, wherein the concentration of M1P is measured ar the trough level.

[0083] In some embodiments, the disclosed composotions of M1P are administered at a dose that achieves a plasma concentraion of greater than 20 pg / mL for at least 20% of time over a dosing cycle (e.g., weekly, or biweekly, or every two weeks). In other embodiments, the disclosed composotions of M1P are administered at a dose that achieves a plasma concentraion of greater than 20 pg / mL for at least 30% of time over a dosing cycle (e.g., weekly, or biweekly, or every two weeks). In other embodiments, the disclosed composotions of M1P are administered at a dose that achieves a plasma concentraion of greater than 20 pg / mL for at least 40% of time over a dosing cycle (e.g., weekly, or biweekly, or every two weeks). In other embodiments, the disclosed composotions of M1P are administered at a dose that achieves a plasma concentraion of greater than 20 pg / mL for at22 sf-6569471PCT / US25 / 17678 27 February 2025 (27.02.2025)Attorney Docket No.: 74220-20015.40 least 50% of time over a dosing cycle (e.g., weekly, or biweekly, or every two weeks). In other embodiments, the disclosed composotions of M1P are administered at a dose that achieves a plasma concentraion of greater than 20 pg / mL for at least 60% of time over a dosing cycle (e.g., weekly, or biweekly, or every two weeks). In other embodiments, the disclosed composotions of M1P are administered at a dose that achieves a plasma concentraion of greater than 20 pg / mL for at least 70% of time over a dosing cycle (e.g., weekly, or biweekly, or every two weeks). In other embodiments, the disclosed composotions of M1P are administered at a dose that achieves a plasma concentraion of greater than 20 pg / mL for at least 80% of time over a dosing cycle (e.g., weekly, or biweekly, or every two weeks). In other embodiments, the disclosed composotions of M1P are administered at a dose that achieves a plasma concentraion of greater than 20 pg / mL for at least 90% of time over a dosing cycle (e.g., weekly, or biweekly, or every two weeks). In other embodiments, the disclosed composotions of M1P are administered at a dose that achieves a plasma concentraion of greater than 20 pg / mL from about 20% to about 90% of time over a dosing cycle (e.g., weekly, or biweekly, or every two weeks). In other embodiments, the disclosed composotions of M1P are administered at a dose that achieves a plasma concentraion of greater than 20 pg / mL from about 30% to about 85% of time over a dosing cycle (e.g., weekly, or biweekly, or every two weeks). In other embodiments, the disclosed composotions of M1P are administered at a dose that achieves a plasma concentraion of greater than 20 pg / mL from about 50% to about 80% of time over a dosing cycle (e.g., weekly, or biweekly, or every two weeks).

[0084] In some embodiments, the disclosed composotions of M1P are administered at a dose that achieves a plasma concentraion of greater than 40 pg / mL for at least 20% of time over a dosing cycle (e.g., weekly, or biweekly, or every two weeks). In other embodiments, the disclosed composotions of M1P are administered at a dose that achieves a plasma concentraion of greater than 40 pg / mL for at least 30% of time over a dosing cycle (e.g., weekly, or biweekly, or every two weeks). In other embodiments, the disclosed composotions of M1P are administered at a dose that achieves a plasma concentraion of greater than 40 pg / mL for at least 40% of time over a dosing cycle (e.g., weekly, or biweekly, or every two weeks). In other embodiments, the disclosed composotions of M1P are administered at a dose that achieves a plasma concentraion of greater than 40 pg / mL for at least 50% of time over a dosing cycle (e.g., weekly, or biweekly, or every two weeks). In other embodiments, the disclosed composotions of M1P are administered at a dose that23 sf-6569471PCT / US25 / 17678 27 February 2025 (27.02.2025)Attorney Docket No.: 74220-20015.40 achieves a plasma concentraion of greater than 40 pg / mL for at least 60% of time over a dosing cycle (e.g., weekly, or biweekly, or every two weeks). In other embodiments, the disclosed composotions of M1P are administered at a dose that achieves a plasma concentraion of greater than 40 pg / mL for at least 70% of time over a dosing cycle (e.g., weekly, or biweekly, or every two weeks). In other embodiments, the disclosed composotions of M1P are administered at a dose that achieves a plasma concentraion of greater than 40 pg / mL for at least 80% of time over a dosing cycle (e.g., weekly, or biweekly, or every two weeks). In other embodiments, the disclosed composotions of M1P are administered at a dose that achieves a plasma concentraion of greater than 40 pg / mL for at least 90% of time over a dosing cycle (e.g., weekly, or biweekly, or every two weeks). In other embodiments, the disclosed composotions of M1P are administered at a dose that achieves a plasma concentraion of greater than 40 pg / mL from about 20% to about 90% of time over a dosing cycle (e.g., weekly, or biweekly, or every two weeks). In other embodiments, the disclosed composotions of M1P are administered at a dose that achieves a plasma concentraion of greater than 40 pg / mL from about 30% to about 85% of time over a dosing cycle (e.g., weekly, or biweekly, or every two weeks). In other embodiments, the disclosed composotions of M1P are administered at a dose that achieves a plasma concentraion of greater than 40 pg / mL from about 50% to about 80% of time over a dosing cycle (e.g., weekly, or biweekly, or every two weeks).

[0085] In any of the foreoing embodiments, the half-life (T1 / 2) of M1P following administration of a composition disclosed herein is from about 50 hous to about 175 hours. In some embodiments, the half-life of M1P is from about 75 hours to about 150 hours. In some embodiments, the half-life of M1P is from about 70 hours to about 120 hours.

[0086] It has been found that the compositions disclosed herein can be administered safely and efficaciously to humans in need thereof. For instance, the compositions can effectively restore the levels of GDP -mannose in the human by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, or at least 90%. In some embodiments, the compositions restore the levels of GDP -mannose in the human of from about 20% to about 80%. In other embodiments, the compositions restore the levels of GDP-mannose in the human of from about 30% to about 70%. In other embodiments, the compositions restore the levels of GDP-mannose in the of from about 40% to about 60%.24 sf-6569471PCT / US25 / 17678 27 February 2025 (27.02.2025)Attorney Docket No.: 74220-20015.40

[0087] In some embodiments, the compositions are administered to a human, including specifically an adult human, weekly or every other week. Administration of the liposomal compositions provided herein can be continuous or intermittent, depending, for example, on the recipient’s physiological condition, whether the purpose of the administration is therapeutic or prophylactic, and other factors known to skilled practitioners. It is within the scope of the present disclosure that different formulations will be effective for different treatments and different disorders, and that administration intended to treat a specific organ or tissue may necessitate delivery in a manner different from that to another organ or tissue. Moreover, dosages may be administered by one or more separate administrations, or by continuous infusion. For repeated administrations over several days or longer, depending on the condition, the treatment is sustained until a desired suppression of disease symptoms occurs. In some variations, the liposomal compositions provided herein may be chronically or intermittently administered to a subject (including, for example, a human) in need thereof. In certain variations, chronic administration is administration of the medicament(s) in a continuous as opposed to acute mode, so as to maintain the initial therapeutic effect (activity) for an extended period of time. In certain variations, intermittent administration is treatment that is not consecutively done without interruption, but rather is cyclic in nature.Liposomal Compositions

[0088] In some embodiments, the liposomal compositions used in the methods herein comprise: at one liposome, wherein each liposome has a liposomal layer and a central core. The M1P or a pharmaceutically acceptable salt thereof is encapsulated within the central core of each liposome. In some embodiments, each liposome has a size between about 70 nM and about 130 nM, or about 100 nM. In some variations of the foregoing, the liposome layer comprises at least three phospholipids. In one variation, the liposome layer comprises 1- dioleoyl-sn-glycero-3 -phosphoethanolamine (DOPE), l,2-dioleoyl-sn-glycero-3- phosphocholine (DOPC), and l,2-distearoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy(polyethylene glycol)-2000] (DSPE-PEG2000).

[0089] The liposomes in the liposomal compositions used in the methods herein may be described in other ways. In other embodiments, the liposomes have a lipid membrane enclosing an intraliposomal compartment, wherein the liposomes encapsulate in the intraliposomal compartment M1P or a pharmaceutically acceptable salt thereof; intraliposomal buffer; extraliposomal buffer; and optionally a radical scavenging antioxidant.25 sf-6569471PCT / US25 / 17678 27 February 2025 (27.02.2025)Attorney Docket No.: 74220-20015.40In some embodiments, the liposomes have an interior surface in contact with the intraliposomal buffer and an external surface in contact with the extraliposomal buffer. In some embodiments the interior and exterior surfaces are in both in contact with a buffer solution of neutral pH.Mannose 1-Phosphate

[0090] In some variations, a pharmaceutically acceptable salt of MIP is present in the liposomes of the liposomal compositions. In certain variations, the pharmaceutically acceptable salt is a dipotassium salt.

[0091] In certain variations, the liposomes comprise a-D(+)mannose 1-phosphate or a pharmaceutically acceptable salt thereof. In one variation, the liposomes comprise substantially pure a-D(+)mannose 1-phosphate or a pharmaceutically acceptable salt thereof.

[0092] In certain variations, the liposomes comprise a-D(+)mannose 1-phosphate dipotassium salt. In certain variations, the liposomes comprise substantially pure a- D(+)mannose 1-phosphate dipotassium salt.

[0093] In some variations of the foregoing, the liposomes has less than about 10%, less than about 5%, less than about 4%, less than about 3%, less than about 2% or less than about 1% of other isomers of MIP, including the beta isomer of MIP.Phospholipids

[0094] In some embodiments, the lipid membrane comprises: (a) at least one phospholipid having an ethanolamine head group and at least one unsaturated fatty acid tail; (b) at least one phospholipid having a choline group and at least one unsaturated fatty acid tail; and (c) at least one phospholipid having an ethanolamine head group and at least one saturated fatty acid tail, conjugated to polyethylene glycol (PEG).

[0095] In certain embodiments with respect the phospholipid having an ethanolamine head group in (a) of the lipid membrane, the unsaturated fatty acid tail independently comprises at least one Cio-28 carbon chain. In certain embodiments, the unsaturated fatty acid tail is an optionally substituted Cio-28 alkenyl or an optionally substituted Cio-28 alkynyl. In some embodiments, each of the fatty acid chains is an unsubstituted Cio-28 alkenyl. In some embodiments, the alkenyl is linear or branched. In some embodiments, each of the fatty acid26 sf-6569471PCT / US25 / 17678 27 February 2025 (27.02.2025)Attorney Docket No.: 74220-20015.40 chains has one or more double bonds. In some embodiments, each double bond has cis configuration. In some embodiments, each double bond has trans configuration. In some embodiments, each of the fatty acid chains is a Cio-28 alkenyl substituted by a substituent selected form the group consisting of acyl, hydroxyl, cycloalkyl, alkoxy, acyloxy, amino, aminoacyl, nitro, halo, thiol, thioalkyl, alkyl, alkenyl, alkynyl and heterocyclyl. In some variations, the phospholipid having an ethanolamine head group in (a) of the lipid membrane has oleoyl tail groups.

[0096] In some embodiments, the phospholipid having an ethanolamine head group and at least one unsaturated fatty acid tail is l,2-dioleoyl-sn-glycero-3 -phosphoethanolamine (DOPE) or a salt thereof. In one variation, the phospholipid having an ethanolamine head group and at least one unsaturated fatty acid tail is:or a salt thereof.

[0097] In certain embodiments with respect to the phospholipid having a choline group in (b) of the lipid membrane, the unsaturated fatty acid tail independently comprises at least one Cio-28 carbon chain. In certain embodiments, the unsaturated fatty acid tail is an unsubstituted Cio-28 alkenyl. In some embodiments, the alkenyl is linear or branched. In some embodiments, the unsaturated fatty acid tail has one or more double bonds. In some embodiments, each double bond has cis configuration. In some embodiments, each double bond has trans configuration. In some embodiments, the unsaturated fatty acid tail is a Cio-28 alkenyl substituted by a substituent selected form the group consisting of acyl, hydroxyl, cycloalkyl, alkoxy, acyloxy, amino, aminoacyl, nitro, halo, thiol, thioalkyl, alkyl, alkenyl, alkynyl and heterocyclyl. In some variations, the phospholipid having choline group in (b) of the lipid membrane has oleoyl tail groups.

[0098] In some variations, the phospholipid having a choline group and at least one unsaturated fatty acid tail is l,2-dioleoyl-sn-glycero-3 -phosphocholine (DOPC), or a salt27 sf-6569471PCT / US25 / 17678 27 February 2025 (27.02.2025)Attorney Docket No.: 74220-20015.40 thereof. In some variations, the phospholipid having a choline group and at least one unsaturated fatty acid tail is:or a salt thereof.

[0099] In some embodiments with respect to the phospholipid having an ethanolamine head group in (c) of the lipid membrane, the saturated fatty acid tail independently comprises at least one C4-28 carbon chain. In certain embodiments, the saturated fatty acid tail is an optionally substituted alkyl. In some embodiments, the saturated fatty acid tail is an unsubstituted C4-28 alkyl. In some embodiments, the saturated fatty acid tail is a C4-28 alkyl substituted by a substituent selected form the group consisting of acyl, hydroxyl, cycloalkyl, alkoxy, acyloxy, amino, aminoacyl, nitro, halo, thiol, thioalkyl, alkyl, alkenyl, alkynyl and heterocyclyl. In some variations, the phospholipid having an ethanolamine head group in (c) of the lipid membrane has stearoyl tail groups.

[0100] In some variations, the phospholipid having an ethanolamine head group and at least one saturated fatty acid tail is l,2-distearoyl-sn-glycero-3 -phosphoethanolamine (DSPE), or a salt thereof. In some variations, the phospholipid having an ethanolamine head group and at least one saturated fatty acid tail is:or a salt thereof.

[0101] In certain variations, the phospholipid conjugated to PEG is DSPE or a salt thereof. In some embodiments, PEGylated phospholipid is DSPE-PEG. In some28 sf-6569471PCT / US25 / 17678 27 February 2025 (27.02.2025)Attorney Docket No.: 74220-20015.40 embodiments, PEGylated phospholipid is DSPE-PEG2000. In some embodiments, the DSPE- PEG is further conjugated to a carbohydrate. In certain embodiments, the DSPE-PEG is further conjugated to a monosaccharide. In some embodiments, the DSPE-PEG is further conjugated to a galactose moiety. In certain embodiments, the DSPE-PEG-galactose has the following structure:or a salt thereof.

[0102] In one embodiment, the lipid membrane comprises DOPC, DOPE and DSPE conjugated to PEG. In one variation, the lipid membrane comprises DOPC:DOPC:DSPE- PEG2000 at a molar ratio of from about 58 : 38 : 3 to 75 : 22 : 3; or from about 48.5 : 48.5 : 3 to about 80 : 17 : 3. In one embodiment, the lipid membrane comprises DOPC:DOPE:DSPE- PEG2000 at a molar ratio of about 58 : 39 : 3. In one embodiment, the lipid membrane comprises DOPC:DOPE:DSPE-PEG2ooo at a molar ratio of about 67 : 30 : 3.

[0103] In certain variations, PEG is present in the composition at a concentration that ranges from about 0.5 molar percent to about 20 molar percent. In some embodiments, PEG is present in the composition at a concentration of about 0.5 molar percent, about 1 molar percent, about 2 molar percent, about 3 molar percent, about 4 molar percent, about 5 molar percent, about 6 molar percent, about 7 molar percent, about 8 molar percent, about 9 molar percent, about 10 molar percent, about 11 molar percent, about 12 molar percent, about 13 molar percent, about 14 molar percent, about 15 molar percent, about 16 molar percent, about 17 molar percent, about 18 molar percent, about 19 molar percent, or about 20 molar percent. In some embodiments, PEG is present in the composition at a concentration of at least about 0.5 molar percent, at least about 1 molar percent, at least about 2 molar percent, at least about 3 molar percent, at least about 4 molar percent, at least about 5 molar percent, at least about 6 molar percent, at least about 7 molar percent, at least about 8 molar percent, at least about 9 molar percent, at least about 10 molar percent, at least about 11 molar percent, at least about 12 molar percent, at least about 13 molar percent, at least about 14 molar percent, at least about 15 molar percent, at least about 16 molar percent, at least about 17 molar percent, at29 sf-6569471PCT / US25 / 17678 27 February 2025 (27.02.2025)Attorney Docket No.: 74220-20015.40 least about 18 molar percent, at least about 19 molar percent, or at least about 20 molar percent; or between 0.5 molar percent and 50 molar percent, between 0.5 molar percent and 40 molar percent, between 0.5 molar percent and 30 molar percent, or between 0.5 molar percent and 20 molar percent.

[0104] In certain variations, PEG is present in the composition at a molecular weight that ranges from about 200 Da to about 40,000 Da. In some embodiments, PEG is present in the composition at a molecular weight that ranges from about 200 Da to about 10,000 Da. In some embodiments, PEG is present in the composition at a molecular weight of about 200 Da, about 300 Da, about 400 Da, about 500 Da, about 600 Da, about 700 Da, about 800 Da, about 900 Da, about 1,000 Da, about 1,500 Da, about 2,000 Da, about 2,500 Da, about 3,000 Da, about 3,500 Da, about 4,000 Da, about 4,500 Da, about 5,000 Da, about 5,500 Da, about 6,000 Da, about 6,500 Da, about 7,000 Da, about 7,500 Da, about 8,000 Da, about 8,500 Da, about 9,000 Da, about 9,500 Da, or about 10,000 Da; or between about 200 Da and about 10,000 Da.Buffers

[0105] In some aspects, provided are compositions that include liposomes having a lipid membrane enclosing an intraliposomal compartment, in which the liposomes encapsulate a mannose phosphate, such as M1P, in the intraliposomal compartment. The compositions further include an intraliposomal buffer comprising a buffering gent, and optionally an acid or a base. In some variations, the pH of the intraliposomal buffer is from about 6.0 to about 7.9. In some embodiments, the pH of the intraliposomal buffer is from about 6.2 to about7.4. In some embodiments, the pH of the intraliposomal buffer is from about 6.4 to about7.5. In some embodiments, the pH of the intraliposomal buffer is from about 6.5 to about7.2. In some embodiments, the pH of the intraliposomal buffer is from about 6.8 to about7.2. In some embodiments, the pH of the intraliposomal buffer is about 6.5. In some embodiments, the pH of the intraliposomal buffer is about 6.8. In some embodiments, the pH of the intraliposomal buffer is about 7.0. In some embodiments, the pH of the intraliposomal buffer is about 7.2. In some embodiments, the pH of the intraliposomal buffer is about 7.3. In some embodiments, the pH of the intraliposomal buffer is about 7.4. In some embodiments, the pH of the intraliposomal buffer is about 7.5.30 sf-6569471PCT / US25 / 17678 27 February 2025 (27.02.2025)Attorney Docket No.: 74220-20015.40

[0106] In some embodiments, the intraliposomal buffer comprises tri s(hydroxymethyl)aminom ethane (Tris). In some embodiments, the intraliposomal buffer comprises 4-(2 -Hydroxy ethyl)- 1 -piperazineethanesulfonic acid (HEPES). In some embodiments, the intraliposomal buffer comprises bicarbonate. In some embodiments, the Tris or HEPES buffer maintain the pH of the intraliposomal compartment at from about 6.8 to about 7.6, or from about 7.0 to about 7.4. In some embodiments, the pH of the intraliposomal buffer is about 7.0. In some embodiments, the pH of the intraliposomal buffer is about 7.2. In some embodiments, the pH of the intraliposomal buffer is about 7.3. In some embodiments, the pH of the intraliposomal buffer is about 7.4. In some embodiments, the pH of the intraliposomal buffer is about 7.5.

[0107] In some embodiments, the intraliposomal buffer comprises a histidine or a citrate buffer. In some such embodiments, the histidine or citrate buffer maintains the pH of the intraliposomal compartment from about 6 to about 7.2, or at a pH of from about 6.2 to about 7.0, or from about 6.4 to about 6.8.

[0108] In some embodiments, the intraliposomal buffer comprises an acetate, ascorbate, benzoate, diethanolamine, phosphate, succinate, tartrate or tromethamine buffering agent..

[0109] In some embodiments, the concentration of the buffering agent (e.g., Tris) in the intraliposomal compartment of the liposomes is at least 15 mM. In other embodiments, the concentration of the buffering agent (e.g., Tris) in the intraliposomal compartment of the liposomes is at least 35 mM. In other embodiments, the concentration of the buffering agent (e.g., Tris) in the intraliposomal compartment of the liposomes is at least 50 mM. In other embodiments, the concentration of the buffering agent (e.g., Tris) in the intraliposomal compartment of the liposomes is at least 50 mM. In other embodiments, the concentration of the buffering agent (e.g., Tris) in the intraliposomal compartment of the liposomes is from about 15 mM to about 75 mM. In other embodiments, the concentration of the buffering agent (e.g., Tris) in the intraliposomal compartment of the liposomes is from about 30 mM to about 60 mM. In other embodiments, the concentration of the buffering agent (e.g., Tris) in the intraliposomal compartment of the liposomes is about 40 mM. In other embodiments, the concentration of the buffering agent (e.g., Tris) in the intraliposomal compartment of the liposomes is about 50 mM.31 sf-6569471PCT / US25 / 17678 27 February 2025 (27.02.2025)Attorney Docket No.: 74220-20015.40

[0110] In some embodiments, the pH of the intraliposomal buffer changes to a minimal extent or does not change upon storage of the liposomal composition for 6 months or 2 years at 5°C or at room temperature. In some embodiments, the pH of the intraliposomal buffer changes by less than 0.4 units (e.g., less than 0.3 units, less than 0.2 units, or less than 0.1 unit) upon storage of the liposomal composition for six month or for 2 years at 5°C or at room temperature. For instance, if the starting pH of the intraliposomal buffer is 7.2, the pH of the intraliposomal buffer will be no less than 6.8 and no more than 7.6 following storage of the liposomal composition for up to 2 years at 5 °C or at room temperature.[OHl] In some variations, the buffering agent comprises a buffer salt. In some embodiments, the extraliposomal buffer comprising a buffer salt and a tonicity modifier. In some variations, the pKa of the buffer salt is between 6.5 to 7.5. In some embodiments, the extraliposomal buffer is in a physiological pH range. In one variation, the buffer salt is tri s(hydroxymethyl)aminom ethane (Tris). In other variations, the extraliposomal buffer comprises bicarbonate, Tris, or HEPES, or any combination thereof. In some embodiments, the concentration of the buffering agent in the extraliposomal buffer is from about 10 mM to about 20 mM. In some embodiments, the concentration of the buffering agent in the extraliposomal buffer is from about 15 mM to about 20 mM.

[0112] In some embodiments, the pH of the extraliposomal buffer changes to a minimal extent or does not change upon storage of the liposomal composition for 6 months or 2 years at 5°C or at room temperature. In some embodiments, the pH of the extraliposomal buffer changes by less than 0.4 units (e.g., less than 0.3 units, less than 0.2 units, or less than 0.1 unit) upon storage of the extraliposomal composition for six month or for 2 years at 5°C or at room temperature. For instance, if the starting pH of the extraliposomal buffer is 7.2, the pH of the extraliposomal buffer will be no less than 6.8 and no more than 7.6 following storage of the liposomal composition for up to 2 years at 5 °C or at room temperature.

[0113] In some variations, the tonicity modifier comprises sugar or saline, or a combination thereof. Suitable sugars that may be used for tonicity include, for example, sucrose and dextrose.

[0114] In other variations, the tonicity modifier is an ionic tonicity modifier. In one variation, the tonicity modifier comprises saline. In some embodiments, the osmolality of the liposomal compositions is isotonic. In some embodiments, the osmolality of the liposomal32 sf-6569471PCT / US25 / 17678 27 February 2025 (27.02.2025)Attorney Docket No.: 74220-20015.40 compositions is from about 270 to about 320 mOsm / kg. In some embodiments, the osmolality of the liposomal compositions is from about 290 to about 320 mOsm / kg. In concentration of the tonicity modifier in the extraliposomal buffer is from about 5 mM to about 25 mM, from about 5 mM to about 15 mM, from about 10 mM to about 20 mM, from about 10 mM to about 15 mM, or from about 15 mM to about 20 mM.

[0115] In some embodiments, the M1P and Tris, and optionally an acid, are present in the composition at a ratio suitable to maintain a neutral pH.

[0116] In some variations, the buffer capacity of the intraliposomal solution may be increased to maintain a neutral pH in the presence of mannose- 1 -phosphate (M1P). In some embodiments, the concentration of the intraliposomal buffer is increased to maintain the neutral pH.Radical Scavenging Antioxidant

[0117] In variations, the composition further comprises a radical scavenging antioxidant. Any suitable radical scavenging antioxidants may be used in the liposomal compositions provided herein. In some embodiments, the radical scavenging antioxidant is butylated hydroyanisole (BHA), butylated hydroxytoluene (BHT), or alpha tocopherol. In one variation, the radical scavenging antioxidant is BHT. In other embodiments, the compositions comprises more than one radical scavenging agent. Any combinations of the radical scavenging antioxidants described herein may be used. For example, in one variation, the compositions comprise BHT and alpha tocopherol. It should be understood that, in certain variations, certain of these radical scavenging antioxidants may be present in the liposomal compositions due to their presence in the lipids used (that are commercially available).Properties of the Liposomal Compositions

[0118] The liposomal compositions described herein are optimized for treating diseases and disorders such as congenital disorders of glycosylation (CDG).

[0119] In some embodiments, the composition has a drug-to-lipid (D / L) ratio of at least 0.01, at least 0.1, or at least 0.2. In certain embodiments, the composition has a D / L ratio from about 0.1 to about 2, from about 0.1 to about 1, from about 0.1 to about 0.5, from about33 sf-6569471PCT / US25 / 17678 27 February 2025 (27.02.2025)Attorney Docket No.: 74220-20015.400.1 to about 0.4, from about 0.1 to about 0.3, from about 0.1 to about 0.2, or from about 0.5 to about 1.5. In some variations, the D / L ratio is about 0.1, 0.2, or 0.3. It should be understood that the drug-to-lipid (D / L) ratio refers to the mass ratio of drug to total lipids in a given sample.

[0120] In some variations of the foregoing, the total mannose phosphate (e.g., MlP)concentration in the sample is between 1 mg / ml and 50 mg / ml, between 1 mg / ml and 25 mg / ml, 1 mg / ml and 15 mg / ml, between 5 mg / ml and 25 mg / ml, between 5 mg / ml and 12 mg / ml, or between 6 mg / ml and 10 mg / ml.

[0121] In certain embodiments, the encapsulated D / L ratio refers to the mass ratio of drug encapsulated in the liposome to total lipids in a given sample. In some variations, the encapsulated D / L ratio does not exceed 0.15. In other variations, the encapsulated D / L ratio is between 0.001 and 0.15, between 0.01 and 0.15, between 0.1 to 0.15, between 0.1 and 0.14, between 0.1 and 0.13, between 0.1 and 0.12, or between 0.1 and 0.11. In other variations, the encapsulated D / L ratio is about 0.1 + / - 0.25%, about 0.1 + / - 0.20%, about 0.1 + / - 15%, or about 0.1 + / - 0.1%.

[0122] In some variations of the foregoing, the total encapsulated mannose phosphate (e.g., MlP)concentration in the sample is between 1 mg / ml and 15 mg / ml, between 5 mg / ml and 12 mg / ml, or between 6 mg / ml and 10 mg / ml.

[0123] In some embodiments, the composition has a mannose phosphate (e.g., MlP)concentration (based on the free acid) from about 1 mg / ml to about 10 mg / ml. In certain embodiments, the mannose phosphate (e.g., M1P) concentration is from about 1 mg / ml to about 10 mg / ml, from about 1 mg / ml to about 9 mg / ml, from about 1 mg / ml to about 8 mg / ml, from about 1 mg / ml to about 7 mg / ml, from about 1 mg / ml to about 6 mg / ml, from about 1 mg / ml to about 5 mg / ml, or from about 1 mg / ml to about 4 mg / ml, from about 1 mg / ml to about 3 mg / ml, or from about 1 mg / ml to about 2 mg / ml. In some variations, the mannose phosphate (e.g., M1P) concentration is at least 1 mg / ml. In some variations, the mannose phosphate (e.g., M1P) concentration is about 1 mg / ml, 2 mg / ml, 3 mg / ml, 4 mg / ml, 5 mg / ml, 6 mg / ml, 7 mg / ml, 8 mg / ml, 9 mg / ml, or 10 mg / ml.

[0124] In other embodiments, the composition has a mannose phosphate (e.g., MlP)concentration (based on the free acid) from about Img / mL to 250 mg / ml, between 1 mg / mL and 200 mg / ml, 1 mg / ml to about 100 mg / ml, from about 1 mg / ml to about 7534 sf-6569471PCT / US25 / 17678 27 February 2025 (27.02.2025)Attorney Docket No.: 74220-20015.40 mg / ml, from about 1 mg / ml to about 50 mg / ml, from about 25 mg / ml to about 75 mg / ml, from about 30 mg / ml to about 55 mg / ml, or from about 30 mg / ml to about 50 mg / ml, or from about 40 mg / ml to about 55 mg / ml, or from about 50 mg / ml to about 55 mg / ml. In some variations of the foregoing, the mannose phosphate (e.g., MlP)concentration is the maximum concentration of mannose phosphate (e.g., MlP)in the liposome.

[0125] In some embodiments, the composition minimizes both lipid degradation and liposomal agglomeration. In some variations, the lipid degradation of the composition is less than 10%, less than 9%, less than 8%, less than 7%, less than 6%, less than 5%, less than 4%, less than 3%, less than 2%, less than 1%, less than 0.5% or less than 0.1%.

[0126] In some embodiments, the liposomal compositions of the disclosure show minimal or no degradation of the individual lipids comprising the liposome. As shown in Example 1, the buffered liposomal compositions are far less prone to lipid degradation compared to compositions formulated in unbuffered solutions (e.g., unbuffered saline solutions). In some embodiments, each of the individual lipids of the liposomal composition degrade less than 10% when stored at 5 °C or room temperature for up to 2 years. In some embodiments, each of the individual lipids of the liposomal composition degrade less than 1% when stored at 5°C or room temperature for up to 2 years. In some embodiments, each of the individual lipids of the liposomal composition degrade less than 0.5% when stored at 5°C for up to 2 years. In some embodiments, each of the individual lipids of the liposomal composition degrade less than 0.25% when stored for up to 2 years.

[0127] In some embodiments, the composition has less than 10%, less than 9%, less than 8%, less than 7%, less than 6%, less than 5%, less than 4%, less than 3%, less than 2%, less than 1%, less than 0.5% or less than 0.1% of total lipid impurities.

[0128] The buffered liposomal compositions of the disclosure show high purity levels upon storage for 6 months or more. For instance, in particular embodiments, the total impurity level observed in the liposomal compositions of the disclosure is less than 1%, less than 0.5%, less than 0.25%, or less than 1% following storage at 5°C or at room temperature for up to 2 years.

[0129] In some embodiments, the composition maintains a pH range between 6.5 and 7, 7 and 7.4 or between 7.35 and 7.45. In certain embodiments, the composition maintains a physiological pH range.35 sf-6569471PCT / US25 / 17678 27 February 2025 (27.02.2025)Attorney Docket No.: 74220-20015.40

[0130] In some embodiments, the composition has a Z-average between 80 nm and 130 nm, between 80 nm and 120 nm, between 80 nm and 110 nm, between 80 nm and 100 nm, between 90 nm and 130 nm, between 90 nm and 120 nm, between 90 nm and 110 nm, or between 90 nm and 100 nm.

[0131] In some embodiments, the composition has a poly dispersity index of less than 0.2 or less than 0.1.

[0132] In some embodiments, no free mannose phosphate (e.g., MlP)is detected in the composition.

[0133] In some embodiments, percentage (%) of encapsulated M1P is the percent of encapsulated mannose phosphate (e.g., M1P) divided by the total mannose phosphate in the finished drug product. It should be understood that the finished drug product refers to the liposomal composition. In some variations, the percentage of encapsulated mannose phosphate (e.g., M1P) is at least about 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 99%, or between 90% and 99%, or between about 90% and 95%, or close to 100%.

[0134] In some embodiments, percentage (%) encapsulation efficiency (“%EE”) is the efficiency of mannose phosphate (e.g., M1P) encapsulation in the liposomes e.g., of the finished drug product) relative to the starting amount of mannose phosphate. In some variations, the %EE is between about 5% and 50%, between about 10% and 30%, or between about 10% and 25%.

[0135] The compositions provided herein may have any one or more of the properties described above. For example, in some variations, the composition provided herein has all of the following properties:(i) a lipid degradation of less than 10%;(ii) a Z-average between 80 nm and 130 nm;(iii) a percentage of encapsulated M1P of at least 80%; and(iv) a pH range between 6.8 and 7.4.36 sf-6569471PCT / US25 / 17678 27 February 2025 (27.02.2025)Attorney Docket No.: 74220-20015.40

[0136] In other variations, the composition provided herein has all of the following properties:(i) a lipid degradation of less than 10%;(ii) a Z-average between 80 nm and 130 nm;(iii) a percentage of encapsulated M1P of at least about 75%, or an encapsulated M1P concentration of at least 5 mg / ml;(iv) an encapsulated D / L ratio of between 0.05 and 0.15; and(v) a pH range between 6.8 and 7.4.

[0137] Stability of the liposomal compositions provided herein, as characterized based on for example lipid degradation, total lipid impurities, pH, Z-average, PDI, Total M1P, encapsulation efficiency, and osmolality, may be measured over a time period over a range of temperatures, such as 5°C, 25°C and 40°C. In some variations, the time period is 1-3 months. In other variations, the time period is at least 6 months, at least 1 year, or at least 2 years.

[0138] The liposomal compositions of the disclosure also display negligible leakage of the M1P from the intraliposomal component, even following storage of the liposomal composition for up to 2 years. For instance, in some embodiments, less than 10%, less than 5%, less than 2%, less than 1%, less than 0.5%, less than 0.25%, or less than 0.1% of the intraliposomal mannose phosphate (e.g., M1P) is released from the intraliposomal component of the liposomal composition following storage for up to 2 years at 5°C or at room temperature.Pharmaceutical Compositions

[0139] Liposomal compositions used in the methods described herein can be incorporated into a variety of formulations for therapeutic use (e.g., by administration) or in the manufacture of a medicament (e.g., for delivering M1P to a subject in need thereof and / or delivering M1P to the cell interior of a subject in need thereof and / or for treating or preventing a disease or disorder such as a congenital disorder of glycosylation (CDG) in a subject in need thereof and / or for treating ataxia in a human suffering from PMM2-CDG) by37 sf-6569471PCT / US25 / 17678 27 February 2025 (27.02.2025)Attorney Docket No.: 74220-20015.40 combining the composition with appropriate carriers (including, for example, pharmaceutically acceptable carriers or diluents), and may be formulated, for example, into preparations in liquid form.

[0140] The components used to formulate the pharmaceutical compositions are preferably of high purity and are substantially free of potentially harmful contaminants, e.g., at least compendial grade, at least United States Pharmacopeia (USP) grade, at least National Food (NF) grade, at least analytical grade, at least pharmaceutical grade or at least suitable for human use. Moreover, compositions intended for in vivo use are sterile. To the extent that a given compound must be synthesized prior to use, the resulting product is typically substantially free of any potentially toxic agents, particularly any endotoxins, which may be present during the synthesis or purification process. Compositions for parental administration are also sterile, substantially isotonic and made under GMP conditions.

[0141] Pharmaceutical compositions of the present disclosure may be used in accord with known methods, such as intravenous administration as a bolus or by continuous infusion over a period of time.

[0142] Dosages and desired concentration of pharmaceutical compositions of the present disclosure may vary depending on the particular use envisioned. The determination of the appropriate dosage or route of administration is well within the skill of an ordinary artisan. Animal experiments provide reliable guidance for the determination of effective doses for human therapy. Interspecies scaling of effective doses can be performed following the principles described in Mordenti, J. and Chappell, W. “The Use of Interspecies Scaling in Toxicokinetics,” In Toxicokinetics and New Drug Development, Yacobi et al., Eds, Pergamon Press, New York 1989, pp.42-46.Articles of Manufacture and Kits

[0143] The present disclosure also provides articles of manufacture and / or kits containing any of the liposomal compositions described herein for use according to the methods to treat CDG as described herein. Articles of manufacture and / or kits of the present disclosure may include one or more containers comprising a purified liposomal composition of the present disclosure. Suitable containers may include, for example, bottles, vials, syringes, and IV solution bags. The containers may be formed from a variety of materials such as glass or plastic. In some embodiments, the articles of manufacture and / or kits further include38 sf-6569471PCT / US25 / 17678 27 February 2025 (27.02.2025)Attorney Docket No.: 74220-20015.40 instructions for use in accordance with any of the methods of the present disclosure. In some embodiments, these instructions comprise a description of administration of any of the liposomal compositions described herein to treat a congenital disorder of glycosylation (CDG) to a subject in need thereof, according to any of the methods of the present disclosure. In some embodiments, the instructions comprise a description of how to detect a congenital disorder of glycosylation (CDG), for example in a subject, in a tissue sample, or in a cell. The article of manufacture and / or kit may further comprise a description of selecting a subject suitable for treatment based on identifying whether that subject has the disease and the stage of the disease.

[0144] The instructions generally include information as to dosage, dosing schedule, and route of administration for the intended treatment. The containers may be unit doses, bulk packages (e.g., multi-dose packages) or sub-unit doses. Instructions supplied in the articles of manufacture and / or kits of the present disclosure are typically written instructions on a label or package insert (e.g., a paper sheet included in the article of manufacture and / or kit), but machine-readable instructions (e.g., instructions carried on a magnetic or optical storage disk) are also acceptable.

[0145] The label or package insert indicates that the composition is used for treating a congenital disorder of glycosylation (CDG). Instructions may be provided for practicing any of the methods described herein.

[0146] The articles of manufacture and / or kits of the present disclosure may be in suitable packaging. Suitable packaging includes, for example, vials, bottles, jars, and flexible packaging (e.g., sealed Mylar or plastic bags). Also contemplated are packages for use in combination with a specific device, such as an inhaler, nasal administration device (e.g., an atomizer) or an infusion device such as a minipump. An article of manufacture and / or kit may have a sterile access port (for example the container may be an intravenous solution bag or a vial having a stopper pierceable by a hypodermic injection needle). The container may also have a sterile access port (e.g, the container may be an intravenous solution bag or a vial having a stopper pierceable by a hypodermic injection needle). At least one active agent in the composition is a carbohydrate such as M1P capable of treating a congenital disorder of glycosylation (CDG) and / or improving one or more symptoms thereof. The container may further comprise a second pharmaceutically active agent.39 sf-6569471PCT / US25 / 17678 27 February 2025 (27.02.2025)Attorney Docket No.: 74220-20015.40

[0147] Articles of manufacture and / or kits may optionally provide additional components such as buffers and interpretive information. Normally, the article of manufacture and / or kit comprises a container and a label or package insert(s) on or associated with the container.ENUMERATED EMBODIMENTS

[0148] The following enumerated embodiments are representative of some aspects of the invention.Al . A method of treating a congenital disorder of glycosylation (CDG) in a human in need thereof, comprising: administering to the human a liposomal composition comprising mannose 1- phosphate or a pharmaceutically acceptable salt thereof, at a dose between 10 mg / kg and 40 mg / kg.A2. The method of embodiment Al, wherein the CDG is phosphomannomutase 2- congenital disorder of glycosylation (PMM2-CDG).A3. The method of embodiment Al or A2, wherein the liposomal composition is administered once weekly.A4. The method of embodiment Al or A2, wherein the liposomal composition is administered once every other week.A5. The method of any one of embodiments Al to A4, wherein the liposomal composition is intravenously.A6. The method of embodiment A5, wherein the liposomal composition is administered via intravenous infusion.A7. The method of any one of embodiments Al to A6, wherein the human is an adult.A8. The method of any one of embodiments Al to A7, wherein the liposomal composition comprises a pharmaceutically acceptable salt of mannose 1 -phosphate.A9. The method of any one of embodiments Al to A8, wherein the liposomal composition comprises a-D(+)mannose 1 -phosphate or a pharmaceutically acceptable salt thereof.40 sf-6569471PCT / US25 / 17678 27 February 2025 (27.02.2025)Attorney Docket No.: 74220-20015.40A10. The method of embodiment A9, wherein the composition comprises substantially pure a-D(+)mannose 1 -phosphate or a pharmaceutically acceptable salt thereof.Al l. The method of any one of embodiments Al to A10, wherein the pharmaceutically acceptable salt is a di-potassium salt.A12. The method of any one of embodiments Al to Al l, wherein the liposomal composition comprises: at one liposome, wherein each liposome has a liposomal layer and a central core; andM1P or a pharmaceutically acceptable salt thereof encapsulated within the central core of each liposome.A13. The method of embodiment A12, wherein each liposome has a size between about 70 nM and about 130 nM.A14. The method of embodiment A13, wherein each liposome has a size of about 100 nM.A15. The method of any one of embodiments A12 to A14, wherein the liposome layer comprises at least three phospholipids.A16. The method of any one of embodiments A12 to A15, wherein the liposome layer comprises l-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), l,2-dioleoyl-sn-glycero-3- phosphocholine (DOPC), and l,2-distearoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy(polyethylene glycol)-2000] (DSPE-PEG2000).Al 7. The method of any one of embodiments Al to Al 1, wherein the liposomal composition comprises: liposomes having a lipid membrane enclosing an intraliposomal compartment, wherein the liposomes encapsulate in the intraliposomal compartment mannose- 1 -phosphate (M1P), and wherein the lipid membrane comprises:(a) at least one phospholipid having an ethanolamine head group and at least one unsaturated fatty acid tail;(b) at least one phospholipid having a choline group and at least one unsaturated fatty acid tail; and41 sf-6569471PCT / US25 / 17678 27 February 2025 (27.02.2025)Attorney Docket No.: 74220-20015.40(c) at least one phospholipid having an ethanolamine head group and at least one saturated fatty acid tail, conjugated to polyethylene glycol (PEG), intraliposomal buffer comprising a buffer salt and optionally acid, wherein the pKa of the buffer salt is between 6 to 8.5; and extraliposomal buffer comprising a buffer salt and a tonicity modifier, wherein the pKa of the buffer salt is between 6 to 8.5.A18. The method of embodiment A17, wherein at least one phospholipid having an ethanolamine head group and at least one unsaturated fatty acid tail is 1,2-dioleoyl-sn- glycero-3 -phosphoethanolamine (DOPE), or a salt thereof.Al 9. The method of embodiment Al 7 or Al 8, wherein at least one phospholipid having a choline group and at least one unsaturated fatty acid tail is l,2-dioleoyl-sn-glycero-3- phosphocholine (DOPC), or a salt thereof.A20. The method of any one of embodiments Al 7 to Al 9, wherein at least one phospholipid having an ethanolamine head group and at least one saturated fatty acid tail is l,2-distearoyl-sn-glycero-3-phosphoethanolamine (DSPE), or a salt thereof.A21. The method of any one of embodiments Al 7 to A20, wherein the lipid membrane comprises DOPC, DOPE and DSPE conjugated to PEG.A22. The method of embodiment A21, wherein the lipid membrane comprises DOPC, DOPE and DSPE-PEG2000.A23. The method of any one of embodiments Al 7 to A22, wherein the drug-to-lipid (D / L) ratio of the composition is at least 0.1.A24. The method of any one of embodiments A17 to A23, wherein the tonicity modifier is an ionic tonicity modifier or sugar.A25. The method of any one of embodiments Al 7 to A23, wherein the tonicity modifier comprises saline.A26. The method of any one of embodiments Al 7 to A25, wherein the intraliposomal comprises Tris.42 sf-6569471PCT / US25 / 17678 27 February 2025 (27.02.2025)Attorney Docket No.: 74220-20015.40A27. The method of any one of embodiments Al 7 to A26, wherein the extraliposomal buffer comprises Tris and saline.A28. The method of any one of embodiments A17 to A26, wherein M1P and Tris, and optionally acid, are present in the composition at a ratio suitable to maintain a neutral pH.A29. The method of any one of embodiments Al 7 to A28, wherein the intraliposomal buffer comprises about 50 mM of Tris; and the extraliposomal buffer comprises 15 mM of Tris and at least 145 mM of saline.A30. The method of any one of embodiments A12 to A29, wherein each liposome further comprises at least one radical scavenging antioxidant.A31. The method of any one of embodiments Al 2 to A30, wherein each liposome further comprises butylated hydroyanisole (BHA), butylated hydroxytoluene (BHT), or alpha tocopherol, or any combination thereof.A32. The method of any one of embodiments A12 to A30, wherein each liposome further comprises butylated hydroxytoluene (BHT).Bl. A method of treating ataxia in a human suffering from phosphomannomutase 2- congenital disorder of glycosylation (PMM2-CDG), the method comprising: administering to the human a liposomal composition comprising mannose 1- phosphate or a pharmaceutically acceptable salt thereof, at a dose between 10 mg / kg and 30 mg / kg of mannose- 1 -phosphate.B2. The method of embodiment Bl, further comprising: improving the human’s (i) postural and gait disturbances, (ii) limb ataxia, (iii) dysarthria, and / or (iv) oculomotor disorders.B3. The method of embodiment Bl, further comprising: periodically measuring an International Cooperative Ataxia Rating Scale (ICARS) score for (i) postural and gait disturbances, (ii) limb ataxia, (iii) dysarthria, and / or (iv) oculomotor disorders during the treatment duration in the human; and assessing the human’s ICARS scores over the treatment duration.43 sf-6569471PCT / US25 / 17678 27 February 2025 (27.02.2025)Attorney Docket No.: 74220-20015.40B4. The method of embodiment Bl, wherein the liposomal composition is administered to the human for a treatment duration over a plurality of weeks, wherein an International Cooperative Ataxia Rating Scale (ICARS) score is measured periodically for (i) postural and gait disturbances, (ii) limb ataxia, (iii) dysarthria, and / or (iv) oculomotor disorders during the treatment duration, and wherein the human’s (i) postural and gait disturbances, (ii) limb ataxia, (iii) dysarthria, and / or (iv) oculomotor disorders improves over the treatment duration based on the ICARS scores.B5. The method of any one of embodiments B 1 to B4, wherein the liposomal composition is administered once weekly.B6. The method of any one of embodiments B 1 to B4, wherein the liposomal composition is administered once every other week.B7. The method of any one of embodiments B 1 to B4, wherein the liposomal composition is intravenously.B8. The method of embodiment B7, wherein the liposomal composition is administered via intravenous infusion.B9. The method of any one of embodiments Bl to B8, wherein the human is an adult.BIO. The method of any one of embodiments B 1 to B8, wherein the human is a child.B 11. The method of any one of embodiments Bl to B 10, wherein the liposomal composition comprises a pharmaceutically acceptable salt of mannose 1 -phosphate.B12. The method of any one of embodiments B 1 to Bl l, wherein the liposomal composition comprises a-D(+)mannose 1 -phosphate or a pharmaceutically acceptable salt thereof.B13. The method of embodiment Bl 2, wherein the composition comprises substantially pure a-D(+)mannose 1 -phosphate or a pharmaceutically acceptable salt thereof.B14. The method of any one of embodiments B 1 to B13, wherein the pharmaceutically acceptable salt is a di-potassium salt.44 sf-6569471PCT / US25 / 17678 27 February 2025 (27.02.2025)Attorney Docket No.: 74220-20015.40Bl 5. The method of any one of embodiments B 1 to B14, wherein the liposomal composition comprises: at one liposome, wherein each liposome has a liposomal layer and a central core; andM1P or a pharmaceutically acceptable salt thereof encapsulated within the central core of each liposome.Bl 6. The method of embodiment Bl 5, wherein each liposome has a size between about 70 nM and about 130 nM.Bl 7. The method of embodiment Bl 6, wherein each liposome has a size of about 100 nM.Bl 8. The method of any one of embodiments B 15 to Bl 7, wherein the liposome layer comprises at least three phospholipids.Bl 9. The method of any one of embodiments B 15 to Bl 8, wherein the liposome layer comprises l-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), l,2-dioleoyl-sn-glycero-3- phosphocholine (DOPC), and l,2-distearoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy(polyethylene glycol)-2000] (DSPE-PEG2000).B20. The method of any one of embodiments Bl to B 19, wherein the liposomal composition comprises: liposomes having a lipid membrane enclosing an intraliposomal compartment, wherein the liposomes encapsulate in the intraliposomal compartment mannose- 1 -phosphate (M1P), and wherein the lipid membrane comprises:(a) at least one phospholipid having an ethanolamine head group and at least one unsaturated fatty acid tail;(b) at least one phospholipid having a choline group and at least one unsaturated fatty acid tail; and(c) at least one phospholipid having an ethanolamine head group and at least one saturated fatty acid tail, conjugated to polyethylene glycol (PEG), intraliposomal buffer comprising a buffer salt and optionally acid, wherein the pKa of the buffer salt is between 6 to 8.5; and45 sf-6569471PCT / US25 / 17678 27 February 2025 (27.02.2025)Attorney Docket No.: 74220-20015.40 extraliposomal buffer comprising a buffer salt and a tonicity modifier, wherein the pKa of the buffer salt is between 6 to 8.5.B21. The method of embodiment B20, wherein at least one phospholipid having an ethanolamine head group and at least one unsaturated fatty acid tail is 1,2-dioleoyl-sn- glycero-3 -phosphoethanolamine (DOPE), or a salt thereof.B22. The method of embodiment B20 or B21, wherein at least one phospholipid having a choline group and at least one unsaturated fatty acid tail is l,2-dioleoyl-sn-glycero-3- phosphocholine (DOPC), or a salt thereof.B23. The method of any one of embodiments B20 to B22, wherein at least one phospholipid having an ethanolamine head group and at least one saturated fatty acid tail is l,2-distearoyl-sn-glycero-3-phosphoethanolamine (DSPE), or a salt thereof.B24. The method of any one of embodiments B20 to B23, wherein the lipid membrane comprises DOPC, DOPE and DSPE conjugated to PEG.B25. The method of embodiment B24, wherein the lipid membrane comprises DOPC, DOPE and DSPE-PEG2000.B26. The method of any one of embodiments B20 to B25, wherein the drug-to-lipid (D / L) ratio of the composition is at least 0.1.B27. The method of any one of embodiments B20 to B26, wherein the tonicity modifier is an ionic tonicity modifier or sugar.B28. The method of any one of embodiments B20 to B27, wherein the tonicity modifier comprises saline.B29. The method of any one of embodiments B20 to B28, wherein the intraliposomal comprises Tris.B30. The method of any one of embodiments B20 to B29, wherein the extraliposomal buffer comprises Tris and saline.B31. The method of any one of embodiments B20 to B30, wherein M1P and Tris, and optionally acid, are present in the composition at a ratio suitable to maintain a neutral pH.46 sf-6569471PCT / US25 / 17678 27 February 2025 (27.02.2025)Attorney Docket No.: 74220-20015.40B32. The method of any one of embodiments B20 to B31, wherein the intraliposomal buffer comprises about 50 mM of Tris; and the extraliposomal buffer comprises 15 mM of Tris and at least 145 mM of saline.B33. The method of any one of embodiments B 15 to B32, wherein each liposome further comprises at least one radical scavenging antioxidant.B34. The method of any one of embodiments B 15 to B33, wherein each liposome further comprises butylated hydroyanisole (BHA), butylated hydroxytoluene (BHT), or alpha tocopherol, or any combination thereof.B35. The method of any one of embodiments B 15 to B33, wherein each liposome further comprises butylated hydroxytoluene (BHT).Cl . A composition comprising liposomes having a lipid membrane enclosing an intraliposomal compartment, wherein the liposomes encapsulate in the intraliposomal compartment mannose- 1 -phosphate (M1P), and wherein the intraliposomal compartment comprises a buffering agent and optionally an acid or base, wherein the pH of the intraliposomal compartment is from about 6.0 to about 7.9.C2. The composition of embodiment Cl, wherein the pH of the intraliposomal compartment is from about 6.2 to about 7.4.C3. The composition of embodiment Cl or C2, wherein the buffering agent in the intraliposomal compartment is tri s(hydroxymethyl)aminom ethane (Tris).C4. The composition of any one of embodiments C1-C3, wherein the buffering agent in the intraliposomal compartment is 4-(2 -Hydroxy ethyl)- 1 -piperazineethanesulfonic acid (HEPES).C5. The composition of any one of embodiments C1-C3, wherein the buffering agent in the intraliposomal compartment is a histidine or citrate buffer.C6. The composition of any one of embodiments C1-C3, wherein the intraliposomal buffer comprises an acetate, ascorbate, benzoate, diethanolamine, phosphate, succinate, tartrate or tromethamine buffering agent.47 sf-6569471PCT / US25 / 17678 27 February 2025 (27.02.2025)Attorney Docket No.: 74220-20015.40C7. The composition of any one of embodiments C1-C6, wherein the liposomes further comprise an extraliposomal component, wherein the extraliposomal compartment comprises a buffering agent and a tonicity modifier, wherein the pH of the extraliposomal compartment is from about 6.0 to about 7.9.C8. The composition of embodiment C7, wherein the pH of the extraliposomal compartment is from about 6.2 to about 7.4.C9. The composition of embodiment C7 or C8, wherein the buffering agent in the intraliposomal compartment is tri s(hydroxymethyl)aminom ethane (Tris).CIO. The composition of any one of embodiments C1-C9, wherein the lipid membrane comprises:(a) at least one phospholipid having an ethanolamine head group and at least one unsaturated fatty acid tail;(b) at least one phospholipid having a choline group and at least one unsaturated fatty acid tail; and(c) at least one phospholipid having an ethanolamine head group and at least one saturated fatty acid tail, conjugated to polyethylene glycol (PEG).Cl 1. The composition of embodiment CIO, wherein at least one phospholipid having an ethanolamine head group and at least one unsaturated fatty acid tail is 1,2-dioleoyl-sn- glycero-3 -phosphoethanolamine (DOPE), or a salt thereof.C12. The composition of embodiment CIO or Cl 1, wherein at least one phospholipid having a choline group and at least one unsaturated fatty acid tail is 1,2-dioleoyl-sn-glycero- 3 -phosphocholine (DOPC), or a salt thereof.C13. The composition of any one of embodiments C10-C12, wherein at least one phospholipid having an ethanolamine head group and at least one saturated fatty acid tail is l,2-distearoyl-sn-glycero-3-phosphoethanolamine (DSPE), or a salt thereof.C14. The composition of any one of embodiments C10-C13, wherein the lipid membrane comprises DOPC, DOPE and DSPE conjugated to PEG.48 sf-6569471PCT / US25 / 17678 27 February 2025 (27.02.2025)Attorney Docket No.: 74220-20015.40Cl 5. The composition of embodiment Cl 4, wherein the lipid membrane comprises DOPC, DOPE and DSPE-PEG2000.Cl 6. The composition of any one of embodiments Cl -Cl 5, wherein the pH of the intraliposomal buffer does not change upon storage of the liposomal composition for up to 2 years at 5°C.Cl 7. The composition of any one of embodiments Cl -Cl 5, wherein the pH of the intraliposomal buffer does not change upon storage of the liposomal composition for up to 6 months at room temperature.Cl 8. The composition of any one of embodiments Cl -Cl 7, wherein all of the lipids comprising the lipid membrane degrade less than 1% when stored at 5°C or room temperature for up to 2 years.Cl 9. The composition of any one of embodiments Cl -Cl 7, wherein less than 10% of the intraliposomal M1P is released from the intraliposomal component of the liposomal composition following storage for up to 2 years at 5°C or at room temperature.C20. The composition of any one of embodiments Cl -Cl 9 wherein, over a period of time at temperatures between 5°C and 40°C, the composition has a Z-average between 80 nm and 130 nm.C21. The composition of any one of embodiments C1-C20, wherein, over a period of time at temperatures between 5°C and 40°C, the composition has a poly dispersity index of less than 0.2 or less than 0.1.C22. The composition of any one of embodiments C1-C21, wherein, over a period of time at temperatures between 5°C and 40°C, no free M1P is detected in the composition.C23. The composition of any one of embodiments C1-C22, wherein, over a period of time at temperatures between 5°C and 40°C, the composition has an encapsulation efficiency of at least 80%.C24. The composition of any one of embodiments C1-C23, wherein, over a period of time at temperatures between 5°C and 40°C, the composition has a mean osmolality within the range of 290 mOsm / kg to 320 mOsm / kg.49 sf-6569471PCT / US25 / 17678 27 February 2025 (27.02.2025)Attorney Docket No.: 74220-20015.40C25. The composition of any one of embodiments C1-C24, further comprising at least one radical scavenging antioxidant.C26. The composition of embodiment C25, wherein at least one radical scavenging antioxidant is butylated hydroyanisole (BHA), butylated hydroxytoluene (BHT), or alpha tocopherol, or any combination thereof.C27. The composition of any one of embodiments C1-C26, further comprising BHT.C28. The composition of embodiment C27, further comprises alpha tocopherol.C29. The composition of any one of embodiments C7-C28, wherein the tonicity modifier is an ionic tonicity modifier or sugar.C30. The composition of any one of embodiments C7-C28, wherein the tonicity modifier comprises saline.C31. A method of treating a congenital disorder of glycosylation in a subject in need thereof, comprising: administering to the subject a composition of any one of embodiments C1-C30.C32. The method of embodiment C31, wherein the congenital disorder of glycosylation is CDG-Ia, CDG-Ib, CDG-Ie, CDG-Ig, CDG-Ii, CDG-Ik, CDG-Io, CDG-Ip, or DPM2-CDG.C33. The method of embodiment C31 or C32, wherein the composition is administered intravenously.DI . A composition comprising liposomes having a lipid membrane enclosing an intraliposomal compartment, wherein the liposomes encapsulate in the intraliposomal compartment mannose- 1 -phosphate (M1P), and wherein the intraliposomal compartment comprises a buffering agent and optionally an acid or base, wherein the pH of the intraliposomal compartment is from about 6.0 to about 7.9.D2. The composition of embodiment DI, wherein the pH of the intraliposomal compartment is from about 6.2 to about 7.4.D3. The composition of embodiment DI or D2, wherein the buffering agent in the intraliposomal compartment comprises tri s(hydroxymethyl)aminom ethane (Tris).50 sf-6569471PCT / US25 / 17678 27 February 2025 (27.02.2025)Attorney Docket No.: 74220-20015.40D4. The composition of embodiment DI or D2, wherein the buffering agent in the intraliposomal compartment comprises 4-(2 -Hydroxy ethyl)- 1 -piperazineethanesulfonic acid (HEPES).D5. The composition of embodiment DI or D2, wherein the buffering agent in the intraliposomal compartment comprises a histidine or citrate buffer.D6. The composition of embodiment DI or D2, wherein the buffering agent in the intraliposomal compartment comprises an acetate, ascorbate, benzoate, diethanolamine, phosphate, succinate, tartrate or tromethamine.D7. The composition of any one of embodiments DI to D6, wherein the liposomes further comprise an extraliposomal component, wherein the extraliposomal compartment comprises a buffering agent and a tonicity modifier, wherein the pH of the extraliposomal compartment is from about 6.0 to about 7.9.D8. The composition of embodiment D7, wherein the pH of the extraliposomal compartment is from about 6.2 to about 7.4.D9. The composition of embodiment D7 or D8, wherein the buffering agent in the intraliposomal compartment and extraliposomal compartment is tri s(hydroxymethyl)aminom ethane (Tris), and optionally, wherein the concentration of the buffering agent in the intraliposomal compartment is higher than the concentration of the buffering agent in the extraliposomal compartment.D10. The composition of any one of embodiments DI to D9, wherein the lipid membrane comprises:(a) at least one phospholipid having an ethanolamine head group and at least one unsaturated fatty acid tail;(b) at least one phospholipid having a choline group and at least one unsaturated fatty acid tail; and(c) at least one phospholipid having an ethanolamine head group and at least one saturated fatty acid tail, conjugated to polyethylene glycol (PEG).51 sf-6569471PCT / US25 / 17678 27 February 2025 (27.02.2025)Attorney Docket No.: 74220-20015.40Dl l. The composition of embodiment DIO, wherein at least one phospholipid having an ethanolamine head group and at least one unsaturated fatty acid tail is 1,2-dioleoyl-sn- glycero-3 -phosphoethanolamine (DOPE), or a salt thereof.D12. The composition of embodiment DIO or DI 1, wherein at least one phospholipid having a choline group and at least one unsaturated fatty acid tail is 1,2-dioleoyl-sn-glycero- 3 -phosphocholine (DOPC), or a salt thereof.D13. The composition of any one of embodiments DIO to DI 2, wherein at least one phospholipid having an ethanolamine head group and at least one saturated fatty acid tail is l,2-distearoyl-sn-glycero-3-phosphoethanolamine (DSPE), or a salt thereof.D14. The composition of any one of embodiments DIO to D13, wherein the lipid membrane comprises DOPC, DOPE, and DSPE conjugated to PEG.DI 5. The composition of embodiment DI 4, wherein the lipid membrane comprisesDOPC, DOPE, and DSPE-PEG2000.DI 6. The composition of any one of embodiments DI to DI 5, wherein all of the lipids comprising the lipid membrane degrade less than about 1% when stored at 5°C or room temperature for up to 2 years.DI 7. The composition of any one of embodiments DI to DI 5, wherein less than 10% of the intraliposomal M1P is released from the intraliposomal component of the liposomal composition following storage for up to 2 years at 5°C or at room temperature.DI 8. The composition of any one of embodiments DI to D17 wherein, over a period of time at temperatures between 5 °C and 40°C, the composition has a Z-average between 80 nm and 130 nm.DI 9. The composition of any one of embodiments DI to DI 8, wherein, over a period of time at temperatures between 5°C and 40°C, the composition has a poly dispersity index of less than 0.2 or less than 0.1.D20. The composition of any one of embodiments DI to DI 9, wherein, over a period of time at temperatures between 5°C and 40°C, no free M1P is detected in the composition.52 sf-6569471PCT / US25 / 17678 27 February 2025 (27.02.2025)Attorney Docket No.: 74220-20015.40D21. The composition of any one of embodiments DI to D20, wherein, over a period of time at temperatures between 5 °C and 40°C, the composition has a mean osmolality within the range of 290 mOsm / kg to 320 mOsm / kg.D22. The composition of any one of embodiments DI to D21, wherein (a) the composition has an encapsulation efficiency between 5% and 50%; or (b) the percentage of encapsulated M1P is at least about 70%, or a combination of (i) and (ii).D23. The composition of any one of embodiments DI to D22, further comprising at least one radical scavenging antioxidant.D24. The composition of any one of embodiments D7 to D23, wherein the tonicity modifier is an ionic tonicity modifier or sugar.D25. The composition of any one of embodiments D7 to D24, wherein the tonicity modifier comprises saline.D26. A composition comprising liposomes having a lipid membrane enclosing an intraliposomal compartment, wherein the liposomes encapsulate in the intraliposomal compartment a mannose phosphate, and wherein the intraliposomal compartment comprises tri s(hydroxymethyl)aminom ethane (Tris), wherein the liposomes further comprise an extraliposomal component, wherein the extraliposomal component comprises Tris and saline, wherein the concentration of Tris in the intraliposomal compartment is higher than the concentration of Tris in the extraliposomal compartment.D27. The composition of embodiment D26, wherein the mannose phosphate is M1P.D28. The composition of embodiment D26 or D27, wherein the Tris present in the intraliposomal compartment is between about 40 mM and 60 mM, and the Tris present in the extraliposomal compartment is between about 10 mM and 20 mM.D29. The composition of any one of embodiments D26 to D28, wherein the molar ratio of Tris in the intraliposomal compartment to extraliposomal compartment is between about 6: 1 and 2 : 1.53 sf-6569471PCT / US25 / 17678 27 February 2025 (27.02.2025)Attorney Docket No.: 74220-20015.40D30. The composition of any one of embodiments D26 to D29, wherein the saline present in the extraliposomal component is at least about 125 mM.D31. A method of treating a congenital disorder of glycosylation in a subject in need thereof, comprising: administering to the subject a composition of any one of embodiments DI to D30.D32. The method of embodiment D31, wherein the congenital disorder of glycosylation is CDG-Ia, CDG-Ib, CDG-Ie, CDG-Ig, CDG-Ii, CDG-Ik, CDG-Io, CDG-Ip, or DPM2-CDG.D33. The method of embodiment D31 or D32, wherein the composition is administered intravenously.EXAMPLES

[0149] The presently disclosed subject matter will be better understood by reference to the following Examples, which are provided as exemplary of the invention, and not by way of limitation.Example 1: A Study Assessing Pharmacodynamics (PD) Activity, Safety, Tolerability, and Pharmacokinetics (PK) of Formulation A Administered Intravenously to Adult Patients with Formulation A

[0150] In order to assess the PD activity, safety, tolerability, and PK of Formulation A in an adult patient with a confirmed diagnosis of PMM2-CDG, a Phase-2 randomized and openlabel study is performed.

[0151] Formulation A includes M1P encapsulated within the central core of liposomes (also known as small unilamellar vesicles) approximately 100 nm in diameter. M1P belongs to the class of chemical entities known as monosaccharide phosphates. The liposome layer is composed of three phospholipids: 1 -di oleoyl-sn-glycero-3 -phosphoethanolamine (DOPE), l,2-dioleoyl-sn-glycero-3 -phosphocholine (DOPC), and l,2-distearoyl-sn-glycero-3- phosphoethanolamine-N-[methoxy(polyethylene glycol)-2000] (DSPE-PEG2000). Among the functions served by the liposome components, DOPE enhances the fusion of liposomes to the cellular membranes, DOPC stabilizes the structure of the liposomes, and DSPE-PEG2000 prolongs the half-life and inhibits non-specific protein binding to the liposomes. Table 1 shows the components of Formulation A.54 sf-6569471PCT / US25 / 17678 27 February 2025 (27.02.2025)Attorney Docket No.: 74220-20015.40Table 1: Components of Formulation A

[0152] Abbreviations in Table 1 : DOPC = l,2-dioleoyl-sn-glycero-3 -phosphocholine;DOPE = l,2-dioleoyl-sn-glycero-3-phosphocholine 1,2- distearoyl-sn-glycero-3- phosphoethanolamine; DSPE-PEG2000 = l,2-distearoyl-sn-glycero-3-phosphoethanolamine- N- [methoxy (polyethylene glycol)-2000]; Formulation A = liposomal M1P drug product (30%:67%:3%); M1P = mannose-1- phosphate; NaCl = sodium chloride

[0153] The study includes a screening period, a twelve-week treatment period, and a thirty-day follow-up period. Adult patients are injected with intravenous infusions of Formulation A at a dose of 10 mg / kg given weekly or 20 mg / kg given weekly. Half of the patients are given the dose of 10 mg / kg given weekly, and the other half are given the dose of 10 mg / kg given weekly. To be eligible for this study, the patients are diagnosed with PMM2- CDG with a genetic test confirmation.

[0154] The treatment period in this study is twelve weeks. The primary outcome measure is to evaluate changes in coagulation and antithrombosis factors, and specifically, changes in antithrombin (AT-III) and Factor XI activity levels are assessed.

[0155] Secondary outcomes are also measured, including, for example:1. the number of patients who experience a clinically significant change in PD activity determined by evaluating changes in hematology assessments,2. the number of patients who experience a clinically significant change in PD activity determined by evaluating changes in blood chemistry,3. the number of patients with treatment-related adverse events assessed by severity and frequency of the adverse events,55 sf-6569471PCT / US25 / 17678 27 February 2025 (27.02.2025)Attorney Docket No.: 74220-20015.404. the number of patients who experience a clinically significant change determined by assessing a change in hematology laboratory testing,5. the number of patients who experience a clinically significant change determined by assessing a change in blood chemistry laboratory testing,6. the number of patients who experience a clinically significant change determined by assessing a change in urinalysis laboratory testing,7. the number of patients who experience a clinically significant change determined by assessing 12 lead electrocardiogram (ECG) QT interval by QTcF.8. vital signs are assessed by respiratory rate (RR), body temperature, blood pressure (systolic and diastolic), pulse,9. plasma concentrations of total M1P are also evaluated to measure Cmax, tmax, AUCo-iast, AUCo-oo, AUCo-tau, CL, Vz, Vss, and ti / 2.Example 2: Safety Studies

[0156] In vitro and in vivo safety pharmacology studies were conducted with Formulation A (see Example 1) as well as with free M1P and empty liposomes in the mouse, rat, and dog. These studies demonstrated that Formulation A had no noteworthy effects on the respiratory, central nervous, or cardiovascular systems, including no evidence of effects on human Ether- a-go-go-Related Gene (hERG) channel function in vitro or on electrocardiogram waveforms in vivo.

[0157] A core battery of GLP safety pharmacology studies was conducted to evaluate the effects of liposomal M1P on the central nervous system (CNS) (Sprague Dawley rats), respiratory system (Sprague Dawley rats), and cardiovascular system in vivo (telemetered beagle dogs) and in vitro hERG channel inhibition in human cells). All in vivo studies were conducted with liposomal M1P administered IV, which is the intended clinical route of administration.

[0158] High-performance liquid chromatography-charged aerosol detection (HPLC- CAD) methods were developed and validated for the quantification of M1P administered as Formulation A or free M1P.56 sf-6569471PCT / US25 / 17678 27 February 2025 (27.02.2025)Attorney Docket No.: 74220-20015.40

[0159] A summary of findings from the study are noted as follows:• Cardiovascular safety in vitro-. One non-GLP and two GLP studies identified IC50 values of >480, >1440, and >780 pg / mL for inhibition of hERG channels by liposomal M1P in vitro. These values are >6.2-fold, >18.5-fold, and >10-fold, respectively, above the concentration of liposomal M1P (78 pg / mL) required to normalize GDP -mannose levels in patient fibroblasts.• Cardiovascular safety in vivo-. Telemetered dogs exhibited an immediate transient hypotensive response (lower systolic, diastolic, mean, and pulse pressure) consistent with (CARP A), which resolved within 1-hour post-dose. No direct effects on body temperature, electrocardiogram (ECG) waveforms, or heart rate were observed, except increases in heart rate compensatory to hypotension.• Respiratory safety in vivo-. No noteworthy effects on any parameters evaluated (respiratory frequency, tidal volume, calculated minute volume) in Sprague Dawley rats.• Central nervous system safety in vivo-. No noteworthy effects on any parameters evaluated (observations for the Irwin test, body temperature, motor activity) in Sprague Dawley rats.

[0160] Collectively, the safety pharmacology studies suggest that there is minimal to no risk of adverse effects on QTc or on CNS or respiratory safety pharmacology endpoints after administration of liposomal M1P. The reversible effect of liposomal M1P on blood pressure noted in dogs is considered consistent with CARP A, a well-known effect of liposomal formulations. Importantly, CARPA is clinically monitorable and can be ameliorated by premedication with anti-histamines and / or by slowing the liposomal M1P IV infusion rate.Example 3: Clinical Pharmacokinetics Results in Study with Formulation A

[0161] A preliminary blinded analysis was performed to obtain estimates of the single- and repeated-dose PK to guide ongoing clinical development. A blinded summary of select PK parameters, as derived by noncompartmental analysis is presented in Table 2.57 sf-6569471PCT / US25 / 17678 27 February 2025 (27.02.2025)Attorney Docket No.: 74220-20015.40Table 2: Blinded PK Parameters following Single doses of M1P in Formulation Ab: Area under the curve from 0, extrapolated to infinity; AUClast: AUC to the last measurable time point; AUC%Extrap: % of the AUC extrapolated to infinity; Cmax: Maximum concentration; NA: Not applicable.

[0162] In the single ascending dose (SAD) portion of the study, M1P PK were evaluable for four subjects in each of the 3 mg / kg and 10 mg / kg dose groups. Subjects having received 0.3 mg / kg were reported to have all analyzed samples as below the lower limit of quantitation (BLQ) and due to infusion related reactions data for only one subject in the 30 mg / kg had sufficient data suitable for PK analysis.

[0163] Following single 3 mg / kg and 10 mg / kg doses, Cmax and AUC normalized for dose were comparable. The arithmetic average estimated half-life for the 3 mg / kg dose was approximately 35 hours, and between 71-73 hours for 10 mg / kg and 30 mg / kg evaluable subjects. The shorter half-life following 3 mg / kg was due to poor characterization of the terminal elimination phase as evidenced by a high extrapolated AUC in these subject (>30% on average).

[0164] In the multiple ascending dose (MAD) portion of the study, Formulation A PK were evaluable for six subjects in each cohort of 3 mg / kg, 10 mg / kg and 20 mg / kg. The PK was characterized following the first, and 3rdof three weekly doses. M1P PK after 3 consecutive weekly doses is summarized in Table 3. FIG. 1 shows plasma concentration of M1P following three different liposomal compositions comprising various doses of M1P (3 mg / kg, 10 mg / kg and 20 mg / kg) after the third dose of a weekly dosing schedule.58 sf-6569471PCT / US25 / 17678 27 February 2025 (27.02.2025)Attorney Docket No.: 74220-20015.40Table 1: Pharmacokinetics of M1P in Phase 1 Study (Formulation A)

[0165] Following repeated doses, the Cmax and AUC over the dosing interval (AUCTAU) were slightly greater than proportional over the range of 3 mg / kg to 20 mg / kg. Accumulation of Cmax ranged from approximately 1.1 -fold to 1.4-fold whereas AUC ranged from approximately 1.4-fold to 1.6-fold as doses increased from 3 mg / kg to 20 mg / kg.Example 4: Effect of Formulation A treatment on protein N-glycosylation of PMM2- CDG patient-derived fibroblasts

[0166] PMM2-CDG is the most common congenital disorder of glycosylation (CDG). Patients with this disease often carry a compound heterozygote mutation of the gene encoding the phosphor-mannose mutase 2 (PMM2) enzyme. PMM2 converts mannose-6- phosphate (M6P) to mannose- 1 -phosphate (M1P), which is a critical upstream metabolite for proper protein N-glycosylation. Therapeutic options for PMM2-CDG patients are limited to management of the disease symptoms as no drug is currently approved by the FDA to treat this disease. Formulation A is a MIP-loaded liposomal formulation being developed as a candidate drug to treat PMM2-CDG. This example describes the effect of Formulation A treatment on protein N-glycosylation of PMM2-CDG patient-derived fibroblasts. It also characterizes the effect of Formulation A on N-glycomics profiles of cell lines derived from six additional individual CDG patients: ALG1-, ALG3-, ALG6-, ALG12-, DPMI-, and DPAGT1-CDG.MATERIAL AND METHODSPreparation of Liposomes59 sf-6569471PCT / US25 / 17678 27 February 2025 (27.02.2025)Attorney Docket No.: 74220-20015.40

[0167] Liposomes were prepared using the thin-film hydration method. Briefly, the lipids were suspended in chloroform and warmed until completely dissolved. The mixture was then dried to a thin-film coating on the flasks. The thin films were hydrated by addition of a solution of neutral buffer containing the M1P, or Sulfo-Cy5.5-COOH (Lumiprobe, Hunt Valley, MD) payload. The lipid / MlP suspension was warmed, vortexed, and sonicated to achieve complete suspension of lipids. The liposome suspension was subsequently extruded to obtain liposomes of an average size of -100 nm. If necessary, the pH of the suspension was adjusted to the target. Unencapsulated payload was removed from the formulation by dialysis. All liposomes were stored at 2-8°C until use.Cell lines and cell culture

[0168] A panel of CDG patient-derived fibroblast cell lines including GM20942, GM20945, GM20956, GM27226, GM27386, GM20944, GM20949, GM20950, GM20101, and the patient derived lymphoblastoid cell line (LCL), GM20941, were obtained. cHealthy dermal fibroblasts, and healthy lymphoblastoid cell line (LCL) were purchased from commercially available sources. All cell lines were cultured in 1640 medium supplemented with 10% fetal bovine serum (FBS), 1% penicillin-streptomycin, and 2 mM L-glutamine, and maintained at 37°C in a 5% CO2 atmosphere until use. ALG1 mutant human fibroblast cells were grown in Dulbecco's Modified Eagle Medium (DMEM) supplemented with 10% fetal bovine serum (FBS), 1% penicillin-streptomycin, and 2 mM L-glutamine and maintained at 37°C in a 5% CO2 atmosphere until use.Cell Viability Assay

[0169] For cell viability experiments the culture medium was exchanged to treatment medium (glucose-free RPMI 1640 supplemented with 10% dialyzed FBS, 1% penicillin / streptomycin, 2 mM L-glutamine and 1 mM glucose). GM20942 cells were treated in triplicate with concentrations of 0.15, 0.3, 0.5, 0.7, and 1.4 mM Formulation A. Control cells were incubated with phosphate-buffered saline (PBS), and empty wells with cell culture media only served as controls for baseline absorbance values. The plates were incubated for 18 hours at 37°C, after which the culture medium was removed, and the cells rinsed with fresh medium. Aliquots (0.2 mg / mL) of the colorimetric formazan dye XTT dissolved in RPMI without phenol red (125 pL / well) and 3 mg / mL of PMS dissolved in PBS at pH 7.4 (62 nL / well) were added to the cells, and the plates were incubated at 37°C for 2 hours. The absorbance at 450 nm was read using a microplate reader.60 sf-6569471PCT / US25 / 17678 27 February 2025 (27.02.2025)Attorney Docket No.: 74220-20015.40Quantification of GDP Mannose levels

[0170] Cells were grown in 10-cm tissue culture dishes. At confluency, the medium was removed, and the cells were treated with Formulation A. Liposomes were diluted to final concentration of 0.5 mM with treatment medium (described above except glucose concentration was changed to 0.5 mM) and added to triplicate dishes for 24 hours. As a positive control, normal fibroblasts were incubated with medium alone for 24 hours. After 24 hours, the medium was aspirated, and the cells were washed with ice-cold 75 mM ammonium carbonate (pH 7.4) and placed on dry ice. Metabolites were extracted by adding 60% methanol (MeOH) in water pre-chilled to -20°C (1.5 mL per dish) and incubating for 15 minutes on dry ice, centrifuged at 13,000 rpm for 5 minutes at 3 °C, and the supernatants were collected. The samples were stored at -80°C until high performance lipid chromatographymass spectrometry (HPLC-MS / MS) analysis.

[0171] All GDP -mannose concentrations were calculated with Prism and AB SCIEX Analyst, both of which are obtained from commercially available sources.Effect of Formulation A treatment on Intercellular Adhesion Molecule (ICAM)l

[0172] PMM2-deficient fibroblasts (GM20942) were seeded in replicate 10-cm tissue culture dishes. At confluency, the growth medium was exchanged for treatment medium described above except glucose concentration was changed to 0.5 mM and 10 ng / mL tumor necrosis factor-a was added.

[0173] Formulation A was diluted in the treatment medium to give final concentrations of 0 (control), 0.015, 0.16, 0.3, 0.5, and 0.7 mM, and added to GM20942 fibroblasts. After a 24- hour incubation, cells were scraped, and washed with cold PBS. Cells were lysed with radioimmuno-precipitation assay (RIP A) buffer (0.3 mL), then centrifuged at 14,000 x g for 15 minutes at 4°C. The supernatants were collected, and the protein concentration measured using a BCA protein assay kit (Thermo Scientific, catalog # 23225). Proteins in the lysates were resolved by SDS-PAGE followed by western blotting using either anti-ICAM-1 or anti- GAPDH primary antibody. Blots were imaged using an Odyssey CLx Imaging System.Effect of Formulation A treatment on overall protein glycosylation

[0174] PMM2-deficient fibroblasts (GM20942) were treated with Formulation A as described above. Following a 24-hour treatment, cells were lysed as described above and cell61 sf-6569471PCT / US25 / 17678 27 February 2025 (27.02.2025)Attorney Docket No.: 74220-20015.40 lysates were resolved by SDS-PAGE followed by western blotting using either anti-mouse glyceraldehyde 3 -phosphate dehydrogenase (GAPDH) antibody or biotinylated Con A. The membrane was then washed and incubated with IRDye 800CW-coupled goat anti-mouse secondary antibody and IRDye 680RD-streptavidin in Intercept antibody diluent buffer, washed and imaged.A-Glycomic Analyses

[0175] Cells were maintained and treated with Formulation A as described above. N- glycan preparation and Quadrupole Time of Flight (QTOF) analysis was performed as described in Chen et al. (Chen et al. 2016). For each glycan structure, the percentage of total glycans in the sample was calculated. The Glycomics profile for each CDG cell line was compared to its healthy counterpart (fibroblasts or LCL cells). Formulation A modulation of N-glycans was calculated and reported as the fold change versus healthy fibroblasts.In vivo and Ex vivo Fluorescence imaging studies

[0176] BALB / c male mice were purchased from Charles River Laboratory (Wilmington, MA, USA) and used between 20 - 25 grams in weight. After an acclimation period, the animals were injected with 5 nmol free Sulfo-Cy5.5 or GLR0052, a liposome formulation with encapsulated Sulfo-Cy5.5-COOH. Before administration, free sulfo-Cy5.5 and GLR0052 were diluted using the dilution buffer (15 mM Tris-HCl + 145 mM NaCl (pH 7.0)) supplied with the test article. Mice were anesthetized briefly under isoflurane and scanned in ventral and dorsal positions prior to dosing and at 30 minutes, 2, 4, 8, 24, and 48 hours postdose (Spectral Instruments Imaging, SPECTRAL Ami HTX Advanced Molecular Imager) to achieve live whole body imaging. They were allowed to regain consciousness and returned to their cage until the next scheduled imaging timepoint. Fluorescence was analyzed using Aura software (Total fluorescence was shown as Photons / s; data not shown).

[0177] For ex vivo organ fluorescence imaging, mice were dosed with vehicle control (15mM Tris-HCl +145 mM NaCl (Ph 7.0), 5 nmol of free Sulfo-Cy5.5 or GLR0052, at a dose volume of 100 L, and euthanized at 4 hours after injection. Animals were anesthetized as stated above and live imaged before euthanasia. All animals were anesthetized under isoflurane and perfused with lx Dulbecco's phosphate-buffered saline (DPBS) until the fluid exiting the right atrium was clear. Immediately after perfusion and harvest, tissues were placed into individual wells of a 12-well plate (lymph nodes x 2, heart, lung, brain, kidneys,62 sf-6569471PCT / US25 / 17678 27 February 2025 (27.02.2025)Attorney Docket No.: 74220-20015.40 liver, spleen, skeletal muscle, small intestine, and colon) and imaged immediately after removal to minimize tissue degradation. Aura software was used to analyze the numerical pixel value and image information (Fluorescence (mean radiance) data shown as Photons / s / cmA2 / sr) .Assessment of Pharmacokinetics (PK)

[0178] Twenty-seven male CD-I mice (3 / timepoint) were administered a single intravenous (IV) dose of Formulation A at 20 mg / kg. At times 0, 1, 2, 4, 8, 12, 24, 48, and 72 hours after dosing, mice were euthanized, and blood was collected into ethylenediaminetetraacetic acid (EDTA) tubes. Samples were processed to plasma and assessed for M1P concentrations using liquid chromatography-mass spectrometry (LC / MS- MS) analysis at Alturas Analytics. Pharmacokinetic parameters were determined for composite concentration-time data using WinNonlin.

[0179] Male Sprague Dawley rats (n = 3) received a single IV dose of Formulation A at a dose of 10 mg / kg. Rats were serial sampled at 1, 2, 4, 8, 12, 24, 48, and 72 hours after administration. At each time point plasma was collected and M1P was measured by LC / MS- MS as described above.

[0180] Male beagle dogs, 3 per group, were administered with three different doses of Formulation A for evaluation of PK parameters, via intravenous infusion using a calibrated infusion pump, followed by a 1ml sterile saline flush. Doses evaluated were 4.8, 25.2, or 46.8 mg / kg, respectively. Blood samples were collected at pre-dose, and then 1.5, 4, 12, 24, 48, and 72 hours after administration of Formulation A. PK parameters were estimated using Phoenix pharmacokinetics software, and with WinNonLin. A non-compartmental approach consistent with the intravenous route of administration was used for parameter estimation.Statistical Analysis

[0181] Data are presented as means ± standard deviation. Statistically significant (P<0.05) differences were tested using t-tests as indicated in the figure legend.RESULTSFormulation A treatment restores GDP-Mannose, total glycoprotein expression and KAM-1 in PMM-2 patient-derived fibroblasts63 sf-6569471PCT / US25 / 17678 27 February 2025 (27.02.2025)Attorney Docket No.: 74220-20015.40

[0182] Fibroblasts derived from PMM2-CDG patients (GM20942) showed a lower intracellular concentration of GDP -mannose (2.2 + / -0.96 ng / mL), compared to healthy (17.3 + / - 5.3 ng / mL) fibroblasts (p=0.0018) (FIG. 2A). To increase production of GDP-mannose in PMM2-deficient cells, we sought to deliver activated mannose, in the form of mannose 1- phosphate (M1P) via liposome encapsulation. Treatment of PMM2-CDG patient fibroblasts with 0.5 mM Formulation A for 24 hours, resulted in a significant increase of intracellular GDP-mannose, reaching comparable levels to those present in normal fibroblasts (FIG. 2B). Restoration of normal levels of GDP-mannose was observed following treatment of four additional PMM2 patient-derived fibroblasts with different genotypes (FIG. 2B). Incubation of Formulation A (0.5 mM) in cells from each of these patient-derived cells with different PMM2 genotypes consistently restored GDP-mannose levels to or above the levels observed in wild type fibroblasts (FIG. 2B). The average intracellular concentrations of GDP- mannose at baseline was 2.16 ± 0.88 ng / mL in untreated PMM2-CDG cells (FIG. 2B). Given the comparable responses observed for Formulation A in the 5 PMM2-CDG patient- derived cells, all follow up studies were conducted using GM20942 cells as a representative in vitro model.

[0183] We then investigated the ability of Formulation A to modulate the N- glycosylation machinery downstream of GDP-mannose synthesis. Initially, Formulation A’s effect on overall glycoprotein levels was assessed by concanavalin A (Con A) binding to total proteins. Compared to healthy dermal fibroblasts (HDF), treatment of GM20942 PMM2- CDG patient fibroblasts (N=3) with 0.3 mM Formulation A increased cellular glycoprotein levels to approximately 94% (quantification data not shown) of that present in normal fibroblasts (FIG. 2C).

[0184] The intracellular adhesion molecule- 1 (ICAM-1) has been shown to be downregulated in PMM2-deficient fibroblasts (He et. al. 2014). We hypothesized that the observed effect of Formulation A on overall protein glycosylation, as shown by Con A blotting (FIG. 2C) should translate to specific glycoproteins such as ICAM-1. As anticipated, treatment of GM20942 patient fibroblasts with increasing concentrations of Formulation A resulted in a dose-dependent increase in ICAM-1 expression (FIG. 2D). We observed increases of ICAM-1 protein expression when the cells were treated with as low as 0.16 mM Formulation A.64 sf-6569471PCT / US25 / 17678 27 February 2025 (27.02.2025)Attorney Docket No.: 74220-20015.40

[0185] Taken together, these data demonstrate that Formulation A treatment overcomes the PMM2 enzyme deficiency in patient-derived cells and restores downstream protein glycosylation.

[0186] To further evaluate the mechanism of N-glycan pathway restoration following Formulation A treatment of PMM2 -deficient fibroblast, N-gly comics profiling was conducted. Additionally, given the beneficial impact of Formulation A treatment on glycoprotein synthesis in PMM2-deficient fibroblasts, N-glycomics profiling was extended to patient derived cells from CDGs other than PMM2.

[0187] Initially, N-glycosylation profiles of total cellular glycoproteins from CDG patient-derived cells were compared to those of healthy counterparts. To this effect, we took advantage of the well-established N-glycomics workflow developed for patient-derived cells grown in culture (Zhang et al. 2016; Chen et al., 2019). Cellular glycomics analyses were carried out with a representative selection of CDG type I cells, including cultured skin fibroblasts from PMM2-, DPMI-, ALG1-, ALG3-, ALG6- and ALG12-CDG patients, as well as lymphoblastoid cell line (LCL) from a DPAGT1-CDG patient. Healthy dermal fibroblasts (HDF) and parental LCL were used as reference. In HDF, asialo-biantennary glycan (Hex5HexNAc4) and its fucosylated counterpart (FuclHex5HexNAc4) are the most abundant complexed asialoglycans, representing around 4.5% and 14.3%, respectively of total detectable N-glycans (FIG. 3). In these cells, the most abundant sialylated complex glycan was the mono-sialo fucosylated Neu5AclFuclHex5NAc4 (5.86%), followed by its non-fucosylated version as well as the di-sialo biantennary complex glycan at 3.36% and 4.27%, respectively (FIG. 3). Although the inter-flask glycosylation variation in LCL was smaller than that in cultured skin fibroblasts, the overall abundance of complex glycans was low (<5% of total glycan amount). The most abundant complex glycan in parental LCL cells was predicted to be the non-fucosylated asialo, mono-galactosylated, hybrid glycan Hex6HexNAc3 at 6.5%. In contrast to plasma N-glycomics profiles which are rich in complex bianteneray sialylated glycans, high mannose glycans are the most common structures associated with cells grown in culture (Kotsias et al. 2019). Our study confirms these findings and shows that high mannose N-glycans, Mano-gGlcNAc? (Man0-Man9) constitute over 50% of the total N-glycans in both HDF and parental LCLs, with increasing65 sf-6569471PCT / US25 / 17678 27 February 2025 (27.02.2025)Attorney Docket No.: 74220-20015.40 abundance from ManO to Man6 and relatively similar levels from Man6 through Man9 (FIG. 3).

[0188] Comparative analysis of N-glycan profiles of multiple CDG patient-derived cell lines, relative to their wildtype counterparts revealed increases of truncated / small high mannose glycans including ManoGlcNAc? (ManO), Manl-5, and reduction of mature / large high mannose (Man 8-9) glycans in PMM2-, DPMI-, ALG3-, and ALG12-CDG cells. These cells represent CDG deficiencies that directly impair the high mannose assembly in ER. The most significant reduction in Man6-7 was observed in ALG3-CDG, consistent with the role of ALG3 in assembling Man6. This reduction was not observed in ALG1-, ALG6-, ALG12-, and DPAGT1-CDG (FIG. 4). In the case of ALG12-CDG, this observation is consistent with the fact that this deficiency specifically impairs the assembling of C branch of the high mannose glycansALG12-CDG however, showed the most pronounced reduction in high mannose N-glycan Man8-9 and a significant accumulation of Man5 (FIG. 4). In DPM1- CDG, due to a deficiency in dolichol-phosphor-mannose (DPM), its cellular glycosylation changes closely resemble those in ALG3 (FIG. 4), which is the first mannosyltransferase that utilizes DPM as substrate. In PMM2-CDG cells, due to mild and generalized deficiencies in both GDP -mannose and DPM, the changes of high mannose glycans were similar to ALG3 and DPMI -CDG. In cells with type I CDGs that do not directly affect high mannose glycan assembling including DPAGT1-CDG and ALG6-CDG, no obvious changes in high mannose glycan abundance were detected. ALG1-CDG with a defect in the first mannosyl transferase showed the overall mildest phenotype of the all the cell types investigated in this study. Interestingly, a mannose deprived tetrasaccharide was detected in these cells /

[0189] The abundance of asialo-biantennary glycan (Hex5HexNAc4 predicted to be GabMansGlcNAc? and its fucosylated counterpart (FuclHex5HexNAc4) was variable between different type I CDG cells (FIG. 4). While there was virtually no difference in the relative abundance of the non-fucosylated version between PMM2-CDG and HDF, the fucosylated counterpart was significantly reduced in PMM2-, ALG1-, ALG6- and ALG12- CDG (FIG. 4).

[0190] Of the cell lines investigated, ALG12-CDG showed the most significant relative depletion of sialylated glycans, including the mono-sialo fucosylated Neu5AclFuclHex5NAc4, its non-fucosylated version, and the di-sialo biantennary complex glycan Neu5Ac2Hex5HexNAc4. PMM2-CDG cells, on the other hand, appear to contain66 sf-6569471PCT / US25 / 17678 27 February 2025 (27.02.2025)Attorney Docket No.: 74220-20015.40 comparable levels to those found in HDF. Having examined all the complexed glycans, including non-fucosylated, fucosylated, and sialylated glycans, similar to the asialo glycan, no consistent pattern was identified. N-glycans from a DPAGT1-CDG line were compared to the normal line derived from one of the unaffected parents (FIG. 4). While the abundance of fucosylated di -sialylated, mono-sialylated and a-galactosylated glycans was reduced in DPAGT1-CDG cells, the abundance of fucosylated a-sialo-biantennary glycan was increased. Thus, the overall under-glycosylation of complexed glycans was difficult to evaluate in cultured type 1 CDG cells. As expected, no generalized change in Golgi associated glycosylation including glcNAcylation, galactosylation, sialylation or fucosylation was observed.Formulation A corrects the abnormal glycosylation pattern in PMM2-CDG cells

[0191] The panel of control or CDG patient-derived cells discussed above were treated with 1 mM Formulation A (N=3) or vehicle (N=3) for 24 hours. For each cell type, the N- glycan profile of Formulation A-treated CDG cells was compared to that of their corresponding untreated control. Of all the cell types investigated, Formulation A treatment had the most pronounced effect on PMM2-CDG fibroblasts as it significantly increased the abundance of mature high mannose glycans, including Man8 and Man9 in PMM2-CDG cells (p<0.01) and reduced the abundance of truncated high mannose Man0-Man5 (p<0.05) (FIGS. 5, 6). The overall effect of Formulation A treatment was a normalization of high mannose glycans close to the levels seen in healthy control, as these treatment groups were statistically equivalent for all high mannose glycans (Man0-Man9). Interestingly, moderate improvement was also observed in DPM1-CDG cells, with Formulation A treatment increasing the abundance of Man8 and Man9 and reducing the abundance of Man0-5, however these differences were generally not statiscally significant (FIG. 6). Although changes of individual glycans are milder in DPMI -CDG, the overall trend / pattern of improvement is consistent with what was observed in PMM2-CDG. We did not observe any significant change in any other CDG cell line upon Formulation A treatment. In particular, the abundance of mannose-deprived tetrasaccharide did not change in ALG1-CDG cells.The liposome formulation used for Formulation A distributes primarily to the liver

[0192] It is accepted that lipid nanoparticle (including liposomes) biodistribution is driven primarily by their lipid content, rather than their payload. With the objective of characterizing the biodistribution of Formulation A-like liposomes, a surrogate formulation67 sf-6569471PCT / US25 / 17678 27 February 2025 (27.02.2025)Attorney Docket No.: 74220-20015.40(GLR0052), with encapsulated M1P substituted with Sulfo-Cy5.5-COOH, was administered to wild type mice in order to enable live animal imaging followed by ex-vivo imaging of dissected tissues at necropsy. BALB / c mice were administered free Sulfo-Cy5.5 (control) or GLR0052 via intravenous injection and whole body live near infra-red fluorescence (NIRF) imaging was performed at various time points post-injection (data not shown). Optimal NIRF signal could be observed at 4 hours when assessed ventrally (FIG. 7A) for GLR0052. While measurable signal was measurable for free Sulfo-Cy5.5, it was at a lower intensity, compared to that in GLR0052 animals. The 4 hour time point was thus used for a subsequent study focused on ex-vivo imaging of tissues collected from animals administered either vehicle control (15 mM Tris-HCl + 145 mM NaCl (pH 7.0)), GLR0052 or free Sulfo-Cy5.5. Appreciable fluorescence signal was detected in several organs / tissues, including lymph nodes, heart, lung, muscle, kidney, spleen, and GI tract (FIG. 7B). For all these tissues however, signal intensity was significantly higher in GLR0052- compared to free dye-treated animals. The highest signal was observed in the liver, lung and spleen (FIGS. 7B, C). These data are in line with previous liposome biodistribution reports (Sercombe et al, 2015) and confirm the liver, spleen and other lymphatic organs as the primary target tissues for our liposome formulation.Formulation A ’s pharmacokinetics profile supports its further development as a drug candidate for PMM2-CDG

[0193] Following IV administration of Formulation A (20 mg / kg) in mice, M1P levels decreased slowly in a monophasic manner with an estimated terminal half-life (t 1 / 2) of approximately 11.2 hours. The volume of distribution was 0.2 L / kg and was lower than the total body of water in mice (~ 0.7 L / kg), suggesting that Formulation A was not extensively distributed in tissues. Systemic clearance of total M1P was 11.2 mL / min / kg and was lower than hepatic blood flow (~ 90 mL / min / kg) suggesting Formulation A was slowly cleared from the body. The Cmax was 142 pg / mL and AUCiast was 1270 hr*pg / mL (FIG. 8A). The low clearance and low volume of distribution are in line with the relatively long half-life for Formulation A in mice following IV administration. Similar desirable PK properties were observed in Sprague-Dawley rats administered IV with Formulation A at the 10 mg / kg dose. In rats, the Cmax was 72.5 pg / ml with a half-life of 16.2 hours achieving a relatively high overall exposure (FIG. 8B). As we continued to develop Formulation A, it was imperative to also characterize its PK profile in a non-rodent species and we selected the dog for this stage of development. IV infusion of Formulation A to male dogs at 4.8, 25.2, or 46.8 mg / kg68 sf-6569471PCT / US25 / 17678 27 February 2025 (27.02.2025)Attorney Docket No.: 74220-20015.40 resulted in systemic exposure to M1P (FIG. 8C). Peak plasma M1P concentrations were observed over a range from 1 to 4 hours post-start of infusion. Systemic clearance ranged from approximately 2 to 7 mL / h / kg and volume of distribution ranged from approximately 80 to 130 mL / kg, resulting in terminal elimination half-life that generally ranged from approximately 8 at the lowest dose to 40 hours at the highest. As a whole, the PK profile of Formulation A across three different species shows a relatively long systemic exposure with dose-proportional increases in circulating concentrations that reach several fold the estimated in vitro efficacy concentration observed in PMM-2 CDG fibroblasts (FIG. 2).Example 5: Phase 2 Dose Escalation Study - PMM2-CDG Patients with Ataxia

[0194] A Phase 2 dose escalation study was conducted in adult (18+ years) PMM2-CDG patients (N=10 treated patients) with Ataxia Telangiectasia (AT) below the lower limit of normal (LLN). Formulation A was administered on a dosing schedule of weekly IV infusions at 10 mg / kg, 20 mg / kg, and 30 mg / kg for 12-24 weeks. Formulation A was demonstrated to be safe and well tolerated, and preliminary efficacy signals were observed at top dose (30 mg / kg). Doses up to 30 mg / kg were also safe and well-tolerated in PMM2-CDG adolescent (12-17 years) patients (N=4).

[0195] A 30 mg / kg dose was selected for a Phase 2, randomized, open-label, 24-week study to assess clinical effects related to ataxia of multiple doses of Formulation A administered intravenously to adult, adolescent, and pediatric (2-11 years) participants with PMM2-CDG. Participants received weekly infusions of Formulation A, or at least 9 infusions in the first 12 weeks of the treatment period (including the first infusion and the infusion at week 12), or at least 18 infusions over 24 weeks (including the first infusion and the week 24 infusion). Fexofenadine was administered approximately 12 (± 2) hours and 3 (± 1) hours before Formulation A infusion.

[0196] The International Cooperative Ataxia Rating Scale (ICARS) was used to quantify impairment levels resulting from ataxia. The ICARS is an outcome measure created to standardize the quantification of impairment due to cerebellar ataxia. The scale is scored out of 100 with 19 items and 4 subscales of postural and gait disturbances, limb ataxia, dysarthria, and oculomotor disorders. Higher scores indicate higher levels of impairment. The ICARS has been validated for use in patients with focal cerebellar lesions and hereditary spinocerebellar and Friedrich's ataxia. In the present study, the ICARS was scored from 069 sf-6569471PCT / US25 / 17678 27 February 2025 (27.02.2025)Attorney Docket No.: 74220-20015.40(best score / normal) to 100 (worst score / most severe impairment) with scores calculated based on the 4 subscales as described in Table 4 below.Table 4: ICARS scores

[0197] Patient baseline ICARS tests were scored during patient screening days (Days -28 to -1) or pre-dose on Day 1. Changes from baseline in ataxia after 12 and 24 weeks of dosing with Formulation A were characterized.

[0198] ICARS scores were quantified in adult subjects administered 30 mg / kg Formulation A over 12 weeks (N=10), and 24 weeks (N=9), and following a washout period at week 27 (4 weeks after last dose of Formulation A), the results of which are shown in FIG. 9A. Significant clinical improvements were observed at 12 and 24 weeks in ataxia (ICARS).

[0199] Changes in ICARS scores at week 12, week 24 and week 27 (washout) for each subject are shown in FIG. 9B and FIG. 9C. Clinical improvements in ICARS were observed at 12 weeks and maintained or further improved at 24 weeks. FIGS. 10A-10B depict the baseline (FIG. 10A) and week 12 (FIG. 10B) results of an Archimedes Spiral test to assess upper-limb coordination in Subject 2007. FIGS. 10C-10D depict the baseline (FIG. 10C) and week 12 (FIG. 10D) results of an Archimedes Spiral test to assess upper-limb coordination in Subject 2008. FIG. 11 shows a comparison of changes in ICARS scores across previously reported studies in PMM2-CDG patients having undergone Formulation A treatment over 12 weeks and 24 weeks. Standard of Care (SOC) treatment averaged an improvement in ICARS scores of 2.6 points over 12 months in patients 5-18 yrs.Acetazolamide clinical studies averaged an improvement in ICARS score of 6 points over 25 weeks in patients 5-21 yrs. Average ICARS score improvement after 12 and 24 weeks of Formulation A treatment was significantly greater than the previously reported results in PMM2-CDG (i.e., SOC and acetazolamide treatments). The total changes in ICARS scores and change in ICARS scores were measured in each of the 4 subscales: postural / gait; limb coordination; dysarthria; and oculomotor. The most significant impact of Formulation A was observed in limb coordination and postural / gait section of ICARS.70 sf-6569471

Claims

1. PCT / US25 / 17678 27 February 2025 (27.02.2025)Attorney Docket No.: 74220-20015.40CLAIMSWhat is claimed is:

1. A method of treating a congenital disorder of glycosylation (CDG) in a human in need thereof, comprising: administering to the human a liposomal composition comprising mannose 1- phosphate or a pharmaceutically acceptable salt thereof, at a dose between 10 mg / kg and 40 mg / kg.

2. The method of claim 1, wherein the CDG is phosphomannomutase 2-congenital disorder of glycosylation (PMM2-CDG).

3. A method of treating ataxia in a human suffering from phosphomannomutase 2- congenital disorder of glycosylation (PMM2-CDG), the method comprising: administering to the human a liposomal composition comprising mannose 1- phosphate or a pharmaceutically acceptable salt thereof, at a dose between 10 mg / kg and 30 mg / kg of mannose- 1 -phosphate.

4. The method of claim 3, further comprising: improving the human’s (i) postural and gait disturbances, (ii) limb ataxia, (iii) dysarthria, and / or (iv) oculomotor disorders.

5. The method of claim 3, further comprising: periodically measuring an International Cooperative Ataxia Rating Scale (ICARS) score for (i) postural and gait disturbances, (ii) limb ataxia, (iii) dysarthria, and / or (iv) oculomotor disorders during the treatment duration in the human; and assessing the human’s ICARS scores over the treatment duration.

6. The method of claim 3, wherein the liposomal composition is administered to the human for a treatment duration over a plurality of weeks,71 sf-6569471PCT / US25 / 17678 27 February 2025 (27.02.2025)Attorney Docket No.: 74220-20015.40 wherein an International Cooperative Ataxia Rating Scale (ICARS) score is measured periodically for (i) postural and gait disturbances, (ii) limb ataxia, (iii) dysarthria, and / or (iv) oculomotor disorders during the treatment duration, and wherein the human’s (i) postural and gait disturbances, (ii) limb ataxia, (iii) dysarthria, and / or (iv) oculomotor disorders improves over the treatment duration based on the ICARS scores.

7. The method of any one of claims 1 to 6, wherein the liposomal composition is administered once weekly.

8. The method of any one of claims 1 to 7, wherein the liposomal composition is administered once every other week.

9. The method of any one of claims 1 to 8, wherein the liposomal composition is intravenously.

10. The method of claim 9, wherein the liposomal composition is administered via intravenous infusion.

11. The method of any one of claims 1 to 10, wherein the human is an adult.

12. The method of any one of claims 1 to 11, wherein the liposomal composition comprises a pharmaceutically acceptable salt of mannose 1 -phosphate.

13. The method of any one of claims 1 to 12, wherein the liposomal composition comprises a-D(+)mannose 1 -phosphate or a pharmaceutically acceptable salt thereof.

14. The method of claim 13, wherein the composition comprises substantially pure a- D(+)mannose 1 -phosphate or a pharmaceutically acceptable salt thereof.

15. The method of any one of claims 1 to 14, wherein the pharmaceutically acceptable salt is a di -potassium salt.

16. The method of any one of claims 1 to 15, wherein the liposomal composition comprises: at one liposome, wherein each liposome has a liposomal layer and a central core; and72 sf-6569471PCT / US25 / 17678 27 February 2025 (27.02.2025)Attorney Docket No.: 74220-20015.40M1P or a pharmaceutically acceptable salt thereof encapsulated within the central core of each liposome.

17. The method of claim 16, wherein each liposome has a size between about 70 nM and about 130 nM.

18. The method of claim 17, wherein each liposome has a size of about 100 nM.

19. The method of any one of claims 16 to 18, wherein the liposome layer comprises at least three phospholipids.

20. The method of any one of claims 16 to 19, wherein the liposome layer comprises 1- dioleoyl-sn-glycero-3 -phosphoethanolamine (DOPE), l,2-dioleoyl-sn-glycero-3- phosphocholine (DOPC), and l,2-distearoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy(polyethylene glycol)-2000] (DSPE-PEG2000).

21. The method of any one of claims 1 to 15, wherein the liposomal composition comprises: liposomes having a lipid membrane enclosing an intraliposomal compartment, wherein the liposomes encapsulate in the intraliposomal compartment mannose- 1 -phosphate (M1P), and wherein the lipid membrane comprises:(a) at least one phospholipid having an ethanolamine head group and at least one unsaturated fatty acid tail;(b) at least one phospholipid having a choline group and at least one unsaturated fatty acid tail; and(c) at least one phospholipid having an ethanolamine head group and at least one saturated fatty acid tail, conjugated to polyethylene glycol (PEG), intraliposomal buffer comprising a buffer salt and optionally acid, wherein the pKa of the buffer salt is between 6 to 8.5; and extraliposomal buffer comprising a buffer salt and a tonicity modifier, wherein the pKa of the buffer salt is between 6 to 8.5.

22. The method of claim 21, wherein at least one phospholipid having an ethanolamine head group and at least one unsaturated fatty acid tail is l,2-dioleoyl-sn-glycero-3- phosphoethanolamine (DOPE), or a salt thereof.73 sf-6569471PCT / US25 / 17678 27 February 2025 (27.02.2025)Attorney Docket No.: 74220-20015.4023. The method of claim 21 or 22, wherein at least one phospholipid having a choline group and at least one unsaturated fatty acid tail is l,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), or a salt thereof.

24. The method of any one of claims 21 to 23, wherein at least one phospholipid having an ethanolamine head group and at least one saturated fatty acid tail is 1,2-distearoyl-sn- glycero-3 -phosphoethanolamine (DSPE), or a salt thereof.

25. The method of any one of claims 21 to 24, wherein the lipid membrane comprises DOPC, DOPE and DSPE conjugated to PEG.

26. The method of claim 25, wherein the lipid membrane comprises DOPC, DOPE and DSPE-PEG2000.

27. The method of any one of claims 21 to 26, wherein the drug-to-lipid (D / L) ratio of the composition is at least 0.1.

28. The method of any one of claims 21 to 27, wherein the tonicity modifier is an ionic tonicity modifier or sugar.

29. The method of any one of claims 21 to 27, wherein the tonicity modifier comprises saline.

30. The method of any one of claims 21 to 29, wherein the intraliposomal comprises Tris.

31. The method of any one of claims 21 to 30, wherein the extraliposomal buffer comprises Tris and saline.

32. The method of any one of claims 21 to 30, wherein M1P and Tris, and optionally acid, are present in the composition at a ratio suitable to maintain a neutral pH.

33. The method of any one of claims 21 to 32, wherein the intraliposomal buffer comprises about 50 mM of Tris; and the extraliposomal buffer comprises 15 mM of Tris and at least 145 mM of saline.74 sf-6569471

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