Formulation for gene therapy vectors

Abstract

The present disclosure is generally related to formulations, methods of preparation, and methods of use thereof. Specifically, the present disclosure related to formulations suitable for use in therapeutic, diagnostic, and research applications. More specifically, the present disclosure generally relates to a formulation comprising: a) a buffer, wherein the buffer is present in an amount ranging between 5 mM and 30 mM; b) one or more salts, wherein the one or more salts are individually present in an amount ranging between 0.1 mM and 200 mM; c) one or more amino acids, wherein the one or more amino acids are individually present in an amount ranging between 1 and 200 mM; and d) a detergent, wherein the detergent is present in an amount ranging between 0.0001% and 1%; and wherein the formulation is at a pH ranging between 7.5 and 8.5.

Description

FORMULATION FOR GENE THERAPY VECTORSRELATED APPLICATIONS

[0001] This application claims priority to U.S. Provisional Patent Application No. 63 / 643409, filed on May 6, 2024, which is hereby incorporated by reference herein in its entirety.INCORPORATION BY REFERENCE OF MATERIAL IN SEQUENCE LISTING

[0002] This application incorporates the material provided in the accompanying XML file entitled SequenceListing_AAVAN102WO.xml, created May 5, 2025, which is 4,096 bytes in size.FIELD

[0003] The present disclosure is generally related to formulations, methods of preparation, and methods of use thereof. More specifically, the present disclosure related to formulations suitable for use in therapeutic, diagnostic, and research applications, in particular formulations for use in gene therapy.BACKGROUND

[0004] Pharmaceutical formulations for parenteral administration, including for intravenous administration are important for the successful delivery of active ingredients, such as gene therapeutics delivered via viral vector(s), including recombinant AAV. The present disclosure provides formulations suitable for this use. The formulations may comprise one or more inactive ingredient and / or one or more additional active ingredient in addition to the viral vectors. In some embodiments, the formulations of the disclosure can be formulated in formulations suitable for administration in a mammalian subject, e.g., a human.SUMMARY

[0005] Some embodiments herein relate to a formulation, the formulation comprising: a) a buffer, wherein the buffer is present in an amount ranging between 5 mM and30 mM; b) one or more salts, wherein the one or more salts are individually present in an amount ranging between 0.1 mM and 250 mM; c) one or more amino acids, wherein the one or more amino acids are individually present in an amount ranging between 1 and 200 mM; and d) a detergent (e.g., surfactant), wherein the detergent is present in an amount ranging between 0.0001% and 1% (e.g., % volume / volume, unless otherwise indicated); and wherein the buffer may comprise, for example, acetate, citrate, histidine, phosphate, succinate, Tris, or any combination thereof. In some embodiments, the pharmaceutical formulation comprises (a) a viral vector(s) comprising at least one polynucleotide encoding a protein of interest, (b) a Tris buffer, (c) magnesium chloride, (d) optionally, sodium chloride, (e) a poloxamer (e.g., pol oxamer 188), and (f) one or more amino acids, such as arginine or proline, wherein the pharmaceutical formulation, optionally does not comprise a preservative. In some embodiments, the buffer comprises Tris. In some embodiments, the buffer comprises Tris-HCl. In some embodiments, the buffer comprises a combination of Tris and Tris-HCl. In some embodiments, the buffer comprises about 10 mM Tris. In some embodiments, the buffer comprises about 20 mM Tris. In some embodiments, the one or more salts comprise MgCh, NaCl, or a combination thereof. In some embodiments, the one or more salts comprise about 1 mM MgCh. In some embodiments, the one or more salts comprise NaCl in an amount ranging from about 50 mM to about 150 mM. In some embodiments, the one or more salts comprise about 57 mM NaCl. In some embodiments, the one or more salts comprise about 61 mM NaCl. In some embodiments, the one or more salts comprise about 75 mM NaCl. In some embodiments, the one or more salts comprise about 135 mMNaCl. In some embodiments, the one or more salt comprises about 150 mM NaCl. In some embodiments, the one or more salt comprises about 1 mM MgCh and NaCl in an amount ranging from about 50 mM to about 150 mM. In some embodiments, the one or more salt comprises about 1 mM MgCh and about 57 mM NaCl. In some embodiments, the one or more salt comprises about 1 mM MgCh and about 61 mM NaCl. In some embodiments, the one or more salt comprises about 1 mM MgCh and about 75 mM NaCl. In some embodiments, the one or more salt comprises about 1 mM MgCh and about 135 mM NaCl. In some embodiments, the one or more salt comprises about 1 mM MgCh and about 150 mM NaCl. In some embodiments, the formulation further comprises water for injection (WFI). In some embodiments, the one or more amino acids increases a stress resistance of the formulation as compared to the stress resistance of the same formulationin the absence of the one or more amino acids. In some embodiments, stress resistance is selected from agitation stress, shear stress, freeze-thaw stress, thermal stress, or any combination therein. In some embodiments, the one or more amino acids increases the agitation stress resistance of the formulation as compared to the agitation stress resistance of the same formulation in the absence of the one or more amino acids. In some embodiments, the one or more amino acids increases the freeze-thaw stress resistance of the formulation as compared to the freeze-thaw stress resistance of the same formulation in the absence of the one or more amino acids. In some embodiments, the one or more amino acids may comprise, for example, alanine, arginine, glutamic acid, glycine, methionine, proline, serine, or any combination thereof. In some embodiments, the one or more amino acids comprises about 50 mM arginine. In some embodiments, the one or more amino acids comprises about 50 mM proline. In some embodiments, the one or more amino acids comprises about 50 mM arginine and about 50 mM proline. In some embodiments, the detergent increases the thermal stress resistance of the formulation as compared to the thermal stress resistance of the same formulation in the absence of the detergent. In some embodiments, the detergent comprises Pl 88, polysorbate 80 (PS80), or a combination thereof. In some embodiments, the detergent comprises about 0.005% P188. In some embodiments, the formulation is at a pH ranging between about 7.5 and 8.5, about 7.7 and 8.3. In some embodiments, the formulation is at a pH of about 8 or is 8.

[0006] Some embodiments herein relate to a formulation, the formulation comprising: a) a buffer, wherein the buffer is present in an amount ranging between 5 mM and 30 mM; b) one or more salts, wherein the one or more salts are individually present in an amount ranging between 0.1 mM and 250 mM; c) one or more amino acids, wherein the one or more amino acids are individually present in an amount ranging between 1 and 200 mM; and d) a detergent, wherein the detergent is present in an amount ranging between 0.0001% and 1%. In some embodiments, the buffer comprises about 20 mM Tris, the one or more salts comprises about 1 mM MgCh, the one or more amino acids comprises about 50 mM arginine and about 50 mM proline, and the detergent comprises about 0.005% Pl 88, wherein the formulation is at a pH of about 8. In some embodiments, the formulation further comprises water for injection (WFI).

[0007] Some embodiments herein relate to a formulation, the formulation comprising: a) a buffer, wherein the buffer is present in an amount ranging between 5 mM and 30 mM; b) one or more salts, wherein the one or more salts are individually present in an amount ranging between 0.1 mM and 250 mM; c) one or more amino acids, wherein the one or more amino acids are individually present in an amount ranging between 1 and 200 mM; and d) a detergent, wherein the detergent is present in an amount ranging between 0.0001% and 1%. In some embodiments, the buffer comprises about 20 mM Tris, the one or more salts comprises about 1 mM MgCh and about 150 mM NaCl, the one or more amino acids comprises about 50 mM arginine and about 50 mM proline, and the detergent comprises about 0.005% P188, wherein the formulation is at a pH of about 8. In some embodiments, the buffer comprises about 20 mM Tris, the one or more salts comprises about 1 mM MgCh and about 57 mM NaCl, the one or more amino acids comprises about 50 mM arginine and about 50 mM proline, and the detergent comprises about 0.005% Pl 88, wherein the formulation is at a pH of about 8. In some embodiments, the formulation further comprises water for injection (WFI).

[0008] Some embodiments herein relate to a formulation, the formulation comprising: a) a buffer, wherein the buffer is present in an amount ranging between 5 mM and 30 mM; b) one or more salts, wherein the one or more salts are individually present in an amount ranging between 0.1 mM and 250 mM; c) arginine and / or proline, wherein the arginine and / or proline are individually present in an amount ranging between 20 and 70 mM; and d) a detergent, wherein the detergent is present in an amount ranging between 0.0001% and 1%; and wherein the formulation is at a pH ranging between 7.5 and 8.5. In some embodiments, the buffer may comprise, for example, acetate, citrate, histidine, phosphate, succinate, Tris, or any combination thereof. In some embodiments, the buffer comprises Tris. In some embodiments, the buffer comprises about 10 mM Tris. In some embodiments, the buffer comprises about 20 mM Tris. In some embodiments, the one or more salt comprises MgCh, NaCl, or a combination thereof. In some embodiments, the one or more salts comprise about 1 mM MgCh. In some embodiments, the one or more salts comprise NaCl in an amount ranging from about 50 mM to about 150 mM. In some embodiments, the one or more salts comprise about 57 mM NaCl. In some embodiments, the one or more salts comprise about 61 mM NaCl. In some embodiments, the one or more salts comprise about 75 mM NaCl. In some embodiments, the one or more salts comprise about 135 mM NaCl. In some embodiments, theone or more salt comprises about 150 mM NaCl. In some embodiments, the one or more salt comprises about 1 mM MgCh and NaCl in an amount ranging from about 50 mM to about 150 mM. In some embodiments, the one or more salt comprises about 1 mM MgCh and about 57 mM NaCl. In some embodiments, the one or more salt comprises about 1 mM MgCh and about 61 mM NaCl. In some embodiments, the one or more salt comprises about 1 mM MgCh and about 75 mM NaCl. In some embodiments, the one or more salt comprises about 1 mM MgCh and about 135 mM NaCl. In some embodiments, the one or more salt comprises about 1 mM MgCh and about 150 mM NaCl. In some embodiments, the formulation further comprises water for injection (WFI). In some embodiments, the arginine and / or proline increases a stress resistance of the formulation as compared to the stress resistance of the same formulation in the absence of the one or more amino acids. In some embodiments, the stress resistance is selected from agitation stress, shear stress, freeze-thaw stress, thermal stress, or any combination therein. In some embodiments, the one or more amino acids increases the agitation stress resistance of the formulation as compared to the agitation stress resistance of the same formulation in the absence of the one or more amino acids. In some embodiments, the one or more amino acids increases the shear stress resistance of the formulation as compared to the shear stress resistance of the same formulation in the absence of the one or more amino acids. In some embodiments, the one or more amino acids increases the freezethaw stress resistance of the formulation as compared to the freeze-thaw stress resistance of the same formulation in the absence of the one or more amino acids. In some embodiments, the one or more amino acids increases the thermal stress resistance of the formulation as compared to the thermal stress resistance of the same formulation in the absence of the one or more amino acids. In some embodiments, the formulation comprises about 50 mM arginine. In some embodiments, the formulation comprises about 50 mM proline. In some embodiments, the formulation comprises about 50 mM arginine and about 50 mM proline. In some embodiments, the detergent increases the thermal stress resistance of the formulation as compared to the thermal stress resistance of the same formulation in the absence of the detergent. In some embodiments, the detergent comprises Pl 88, PS80, or a combination thereof. In some embodiments, the detergent comprises about 0.005% P188. In some embodiments, the formulation is at a pH ranging between about 7.5 and 8.5. In some embodiments, the formulation is at a pH of about 8. In some embodiments, the buffercomprises about 20 mM Tris, the one or more salts comprises about 1 mM MgCh, the one or more amino acids comprises about 50 mM arginine and about 50 mM proline, and the detergent comprises about 0.005% P188, wherein the formulation is at a pH of about 8. In some embodiments, the buffer comprises about 20 mM Tris, the one or more salts comprises about 1 mM MgCh and about 150 mM NaCl, the one or more amino acids comprises about 50 mM arginine and about 50 mM proline, and the detergent comprises about 0.005% P188, wherein the formulation is at a pH of about 8. In some embodiments, the buffer comprises about 20 mM Tris, the one or more salts comprises about 1 mM MgCh and about 57 mM NaCl, the one or more amino acids comprises about 50 mM arginine and about 50 mM proline, and the detergent comprises about 0.005% P188, wherein the formulation is at a pH of about 8. In some embodiments, the formulation further comprises water for injection (WFI).

[0009] Some embodiments herein relate to a formulation, the formulation comprising: (a) about 20 mM Tris, (b) about 1 mM magnesium chloride (MgCh), (c) about 50 mM arginine, (d) about 50 mM proline, and (e) about 0.005% pol oxamer 188 (Pl 88), wherein the formulation is at a pH of about 8. In some embodiments, the formulation further comprises water, e.g., water for injection (WFI).

[0010] Some embodiments herein relate to a formulation, the formulation comprising: (a) about 20 mM Tris and Tris-HCl in combination, (b) about 1 mM MgCh, (c) about 50-250 mM sodium chloride (NaCl), (d) about 50 mM arginine, (e) about 50 mM proline, and (f) about 0.005% Pl 88, wherein the formulation is at a pH of about 8. In some embodiments, the formulation further comprises water for injection (WFI).

[0011] Some embodiments herein relate to a formulation, the formulation comprising: (a) about 20 mM Tris and Tris-HCl in combination, (b) about 1 mM MgCh, (c) about 57 mM sodium chloride (NaCl), (d) about 50 mM arginine, (e) about 50 mM proline, and (f) about 0.005% Pl 88, wherein the formulation is at a pH of about 8. In some embodiments, the formulation further comprises water for injection (WFI).

[0012] Some embodiments herein relate to a formulation, the formulation comprising: (a) about 20 mM Tris and Tris-HCl in combination, (b) about 1 mM MgCh, (c) about 75 mM sodium chloride (NaCl), (d) about 50 mM arginine, (e) about 50 mM proline, and (f) about 0.005% Pl 88, wherein the formulation is at a pH of about 8. In some embodiments, the formulation further comprises water for injection (WFI).

[0013] Some embodiments herein relate to a formulation, the formulation comprising: (a) about 10 mM Tris, (b) about 1 mM magnesium chloride (MgCh), (c) about 50 mM arginine, (d) about 50 mM proline, and (e) about 0.005% pol oxamer 188 (Pl 88), wherein the formulation is at a pH of about 8. In some embodiments, the formulation further comprises water for injection (WFI).

[0014] Some embodiments herein relate to a formulation, the formulation comprising: (a) about 10 mM Tris and Tris-HCl in combination, (b) about 1 mM MgCh, (c) about 50-250 mM sodium chloride (NaCl), (d) about 50 mM arginine, (e) about 50 mM proline, and (f) about 0.005% Pl 88, wherein the formulation is at a pH of about 8. In some embodiments, the formulation further comprises water for injection (WFI).

[0015] Some embodiments herein relate to a formulation, the formulation comprising: (a) about 10 mM Tris and Tris-HCl in combination, (b) about 1 mM MgCk, (c) about 57 mM sodium chloride (NaCl), (d) about 50 mM arginine, (e) about 50 mM proline, and (f) about 0.005% Pl 88, wherein the formulation is at a pH of about 8. In some embodiments, the formulation further comprises water for injection (WFI).

[0016] Some embodiments herein relate to a formulation, the formulation comprising: (a) about 10 mM Tris and Tris-HCl in combination, (b) about 1 mM MgCh, (c) about 75 mM sodium chloride (NaCl), (d) about 50 mM arginine, (e) about 50 mM proline, and (f) about 0.005% P188, wherein the formulation is at a pH of about 8. In some embodiments, the formulation further comprises water for injection (WFI).

[0017] In some embodiments, the formulation further comprises a stabilizer. In some embodiments, the stabilizer may comprise, for example, human serum albumin (HSA), cyclodextrin, sucrose, sorbitol, or any combination thereof. In some embodiments, the stabilizer is present in an amount ranging between 0.1% and 20%. In some embodiments, the stabilizer is present in an amount ranging between 0. 1 mM and 200 mM. In some embodiments, the stabilizer comprises about 180 mM sorbitol. In some embodiments, the formulation remains stable at temperatures ranging between about -80°C to about 20°C, such as -80, -70, - 60, -50, -40, -30, -20, -10, 0, 10, or 20°C, or at a temperature within a range defined by any two of the aforementioned values. In some embodiments, the formulation remains stable at a temperature of about -80°C, or at a temperate of about -20°C. In some embodiments, the formulation remains stable at temperatures ranging between about 2-8°C. In someembodiments, the formulation remains stable at temperatures ranging between about 15-30°C. In some embodiments, the formulation remains stable at temperatures ranging between about 35-45°C. In some embodiments, the formulation remains stable at a temperature of about 45°C. In some embodiments, the formulation remains stable for at least 5 freeze-thaw cycles. In some embodiments, the formulation further comprises an AAV vector. In certain embodiments, the serotype of the AAV capsid is may comprise AAV1, AAV2, AAV2G9, AAV3, AAV3a, AAV3b, AAV3-3, AAV4, AAV4-4, AAV5, AAV6, AAV6.1, AAV6.2, AAV6.1.2, AAV7, AAV7.2, AAV8, AAV9, AAV9.11, AAV9.13, AAV9.16, AAV9.24, AAV9.45, AAV9.47, AAV9.61, AAV9.68, AAV9.84, AAV9.9, AAV10, AAV11, AAV12, AAV16.3, AAV24.1, AAV27.3, AAV42.12, AAV42-lb, AAV42-2, AAV42-3a, AAV42-3b, AAV42-4, AAV42- 5a, AAV42-5b, AAV42-6b, AAV42-8, AAV42-10, AAV42-11, AAV42-12, AAV42-13, AAV42-15, AAV42-aa, AAV43-1, AAV43-12, AAV43-20, AAV43-21, AAV43-23, AAV43- 25, AAV43-5, AAV44.1, AAV44.2, AAV44.5, AAV223.1, AAV223.2, AAV223.4, AAV223.5, AAV223.6, AAV223.7, AAVl-7 / rh.48, AAVl-8 / rh.49, AAV2-15 / rh.62, AAV2- 3 / rh.61, AAV2-4 / rh.5O, AAV2-5 / rh.51, AAV3.1 / hu.6, AAV3.1 / hu.9, AAV3-9 / rh.52, AAV3- 1 l / rh.53, AAV4-8 / r 11.64, AAV4-9 / rh.54, AAV4-19 / rh.55, AAV5-3 / rh.57, AAV5-22 / rh.58, AAV7.3 / hu.7, AAV16.8 / hu.lO, AAV16.12 / hu.l l, AAV29.3 / bb. l, AAV29.5 / bb.2,AAV106.1 / hu.37, AAV114.3 / hu.4O, AAV127.2 / hu.41, AAV127.5 / hu.42, AAV128.3 / hu.44, AAV130.4 / hu.48, AAV145.1 / hu.53, AAV145.5 / hu.54, AAV145.6 / hu.55, AAV161.1O / hu.6O, AAV161.6 / hu.61, AAV33.12 / hu.l7, AAV33.4 / hu.l5, AAV33.8 / hu. l6, AAV52 / hu.l9, AAV52.1 / hu.2O, AAV58.2 / hu.25, AAVA3.3, AAVA3.4, AAVA3.5, AAVA3.7, AAVC1, AAVC2, AAVC5, AAV-DJ, AAV-DJ8, AAVF3, AAVF5, AAVH2, AAVrh.72, AAVhu.8, AAVrh.68, AAVrh.70, AAVpi.l, AAVpi.3, AAVpi.2, AAVrh.60, AAVrh.44, AAVrh.65, AAVrh.55, AAVrh.47, AAVrh.69, AAVrh.45, AAVrh.59, AAVhu.12, AAVH6, AAVLK03, AAVH-l / hu.l, AAVH-5 / hu.3, AAVLG-10 / rh.40, AAVLG-4 / rh.38, AAVLG-9 / hu.39, AAVN721-8 / rh.43, AAVCh.5, AAVCh.5Rl, AAVcy.2, AAVcy.3, AAVcy.4, AAVcy.5, AAVCy.5Rl, AAVCy.5R2, AAVCy.5R3, AAVCy.5R4, AAVcy.6, AAVhu. l, AAVhu.2, AAVhu.3, AAVhu.4, AAVhu.5, AAVhu.6, AAVhu.7, AAVhu.9, AAVhu.10, AAVhu.l l, AAVhu.13, AAVhu.15, AAVhu.16, AAVhu.17, AAVhu.18, AAVhu.20, AAVhu.21, AAVhu.22, AAVhu.23.2, AAVhu.24, AAVhu.25, AAVhu.27, AAVhu.28, AAVhu.29, AAVhu.29R, AAVhu.31, AAVhu.32, AAVhu.34, AAVhu.35, AAVhu.37, AAVhu.39,AAVhu.40, AAVhu.41, AAVhu.42, AAVhu.43, AAVhu.44, AAVhu.44Rl, AAVhu.44R2, AAVhu.44R3, AAVhu.45, AAVhu.46, AAVhu.47, AAVhu.48, AAVhu.48Rl, AAVhu.48R2, AAVhu.48R3, AAVhu.49, AAVhu.51, AAVhu.52, AAVhu.54, AAVhu.55, AAVhu.56, AAVhu.57, AAVhu.58, AAVhu.60, AAVhu.61, AAVhu.63, AAVhu.64, AAVhu.66, AAVhu.67, AAVhu.14 / 9, AAVhu.t 19, AAVrh.2, AAVrh.2R. AAVrh.8, AAVrh.8R, AAVrh.10, AAVrh.12, AAVrh.13, AAVrh.l3R, AAVrh.14, AAVrh.17, AAVrh.18, AAVrh.19, AAVrh.20, AAVrh.21, AAVrh.22, AAVrh.23, AAVrh.24, AAVrh.25, AAVrh.31,AAVrh.32, AAVrh.33, AAVrh.34, AAVrh.35, AAVrh.36, AAVrh.37, AAVrh.37R2, AAVrh.38, AAVrh.39, AAVrh.40, AAVrh.46, AAVrh.48, AAVrh.48.1, AAVrh.48.1.2, AAVrh.48.2, AAVrh.49, AAVrb.51, AAVrh.52, AAVrh.53, AAVrh.54, AAVrh.56, AAVrh.57, AAVrh.58, AAVrh.61, AAVrh.64, AAVrh.64Rl, AAVrh.64R2, AAVrh.67, AAVrh.73, AAVrh.74, AAVrh8R, AAVrh8R A586R mutant, AAVrh8R R533A mutant, AAAV, BAAV, caprine AAV, bovine AAV, ovine AAV, AAVhEl.l, AAVhErl.5, AAVhER1.14, AAVhErl.8, AAVhErl.16, AAVhErl.18, AAVhErl.35, AAVhErl.7, AAVhErl.36, AAVhEr2.29, AAVhEr2.4, AAVhEr2.16, AAVhEr2.30, AAVhEr2.31, AAVhEr2.36, AAVhER1.23, AAVhEr3.1, AAV2.5T, AAV-PAEC, AAV-LK01, AAV- LK02, AAV-LK03, AAV-LK04, AAV-LK05, AAV-LK06, AAV-LK07, AAV-LK08, AAV- LK09, AAV-LK10, AAV-LK11, AAV-LK12, AAV-LK13, AAV-LK14, AAV-LK15, AAV- LK16, AAV-LK17, AAV-LK18, AAV-LK19, AAV-PAEC2, AAV-PAEC4, AAV-PAEC6, AAV-PAEC7, AAV-PAEC8, AAV-PAEC11, AAV-PAEC12, AAV-2-pre-miRNA-101, AAV-8h, AAV-8b, AAV-h, AAV-b, AAV SM 10-2, AAV Shuffle 100-1, AAV Shuffle 100- 3, AAV Shuffle 100-7, AAV Shuffle 10-2, AAV Shuffle 10-6, AAV Shuffle 10-8, AAV Shuffle 100-2, AAV SM 10-1, AAV SM 10-8, AAV SM 100-3, AAV SM 100-10, BNP61 AAV, BNP62 AAV, BNP63 AAV, AAVrh.50, AAVrh.43, AAVrh.62, AAVrb.48, AAVhu.19, AAVhu.l l, AAVhu.53, AAV4-8 / rh.64, AAVLG-9 / hu.39, AAV54.5 / hu.23, AAV54.2 / hu.22, AAV54.7 / hu.24, AAV54.1 / hu.21, AAV54.4R / hu.27, AAV46.2 / hu.28, AAV46.6 / hu.29, AAV128.1 / hu.43, true type AAV (ttAAV), UPENN AAV 10, Japanese AAV 10 serotypes, AAV CBr-7.1, AAV CBr-7.10, AAV CBr-7.2, AAV CBr-7.3, AAV CBr-7.4, AAV CBr-7.5,AAV CBr-7.7, AAV CBr-7.8, AAV CBr-B7.3, AAV CBr-B7.4, AAV CBr-El, AAV CBr-E2, AAV CBr-E3, AAV CBr-E4, AAV CBr-E5, AAV CBr-e5, AAV CBr-E6, AAV CBr-E7, AAV CBr-E8, AAV CHt-1, AAV CHt-2, AAV CHt-3, AAV CHt-6.1, AAV CHt-6.10, AAV CHt-6.5, AAV CHt-6.6, AAV CHt-6.7, AAV CHt-6.8, AAV CHt-Pl, AAV CHt-P2, AAV CHt-P5, AAV CHt-P6, AAV CHt-P8, AAV CHt-P9, AAV CKd-1, AAV CKd-10, AAV CKd-2, AAV CKd-3, AAV CKd-4, AAV CKd-6, AAV CKd-7, AAV CKd-8, AAV CKd-Bl, AAV CKd- B2, AAV CKd-B3, AAV CKd-B4, AAV CKd-B5, AAV CKd-B6, AAV CKd-B7, AAV CKd- B8, AAV CKd-Hl, AAV CKd-H2, AAV CKd-H3, AAV CKd-H4, AAV CKd-H5, AAV CKd- H6, AAV CKd-N3, AAV CKd-N4, AAV CKd-N9, AAV CLg-Fl, AAV CLg-F2, AAV CLg- F3, AAV CLg-F4, AAV CLg-F5, AAV CLg-F6, AAV CLg-F7, AAV CLg-F8, AAV CLv-1, AAV CLv- 1 , AAV Civ 1 - 10, AAV CLv 1 -2, AAV CLv- 12, AAV CLv 1 -3 , AAV CLv- 13 , AAV CLv 1-4, AAV Civ 1-7, AAV Civ 1-8, AAV Civ 1-9, AAV CLv-2, AAV CLv-3, AAV CLv-4, AAV CLv-6, AAV CLv-8, AAV CLv-Dl, AAV CLv-D2, AAV CLv-D3, AAV CLv-D4, AAV CLv-D5, AAV CLv-D6, AAV CLv-D7, AAV CLv-D8, AAV CLv-El, AAV CLv-Kl, AAV CLv-K3, AAV CLv-K6, AAV CLv-L4, AAV CLv-L5, AAV CLv-L6, AAV CLv-Ml, AAV CLv-Ml l, AAV CLv-M2, AAV CLv-M5, AAV CLv-M6, AAV CLv-M7, AAV CLv-M8, AAV CLv-M9, AAV CLv-Rl, AAV CLv-R2, AAV CLv-R3, AAV CLv-R4, AAV CLv-R5, AAV CLv-R6, AAV CLv-R7, AAV CLv-R8, AAV CLv-R9, AAV CSp-1, AAV CSp-10, AAV CSp-11, AAV CSp-2, AAV CSp-3, AAV CSp-4, AAV CSp-6, AAV CSp-7, AAV CSp- 8, AAV CSp-8.10, AAV CSp-8.2, AAV CSp-8.4, AAV CSp-8.5, AAV CSp-8.6, AAV CSp- 8.7, AAV CSp-8.8, AAV CSp-8.9, AAV CSp-9, AAV.hu.48R3, AAV.VR-355, AAV3B, AAV4, AAV5, AAVF1 / HSC1, AAVF11 / HSC11, AAVF12 / HSC12, AAVF13 / HSC13, AAVF14 / HSC14, AAVF15 / HSC15, AAVF16 / HSC16, AAVF17 / HSC17, AAVF2 / HSC2, AAVF3 / HSC3, AAVF4 / HSC4, AAVF5 / HSC5, AAVF6 / HSC6, AAVF7 / HSC7, AAVF8 / HSC8, AAVF9 / HSC9, PHP.B, PHP. A, G2B-26, G2B-13, TH1.1-32, TH1.1-35, rAAV, myotropic AAV variants, AAV-SLB101, AAV-SLB101 variants, MyoAAV and AAVMYO, AAV-DJ, AAV-DJ8.

[0018] In some embodiments, the formulation further comprises an expression cassette. In some embodiments, the expression cassette is a therapeutic expression cassette. In some embodiments, the therapeutic expression cassette encodes a therapeutic protein. In some embodiments, the therapeutic protein may comprise, for example, frataxin, GSD1A, GSD, PRKAG2, BAG3, Dysferlin, Dystrophin (MicroDys), MYBPC-3, Danon / Lamp2, TMEM43, PKP2, LMNA, TNNT2, RBM20, MYH7, RYR2, CAMKII, PRKAG2, CASQ2, RYR2, DM1,GRN, GBA, MSH3, UNCI 3 A, Stathmin-2, SMET, SMN1 , SCN1 A and / or any truncations and / or variations and / or combination(s) thereof.

[0019] Some embodiments herein relate to a method of treatment, the method comprising: identifying a subject as likely to benefit from treatment and administering one or more of the formulations disclosed herein. In some embodiments, the formulation comprises a nucleic acid encoding a therapeutic protein may comprise, for example, frataxin, GSD1A, GSD, PRKAG2, BAG3, MicroDys, Dysferlin, MYBPC-3, Danon / Lamp2, TMEM43, PKP2, LMNA, TNNT2, RBM20, MYH7, RYR2, CAMKII, PRKAG2, CASQ2, RYR2, DM1, GRN, GBA, MSH3, UNC13A, Stathmin-2, SMEI, SMN1, SCN1A and / or any combination(s) thereof.

[0020] Some embodiments herein relate to the use of one or more formulations disclosed herein in the manufacture of a medicament for the treatment of disease. In some embodiments, the formulation comprises a nucleic acid encoding a therapeutic protein may comprise, for example, frataxin, GSD1A, PRKAG2, BAG3, Dysferlin, MicroDys, MYBPC-3, Danon / Lamp2, TMEM43, PKP2, LMNA, TNNT2, RBM20, MYH7, RYR2, CAMKII, PRKAG2, CASQ2, DM1, GRN, GBA, MSH3, UNC13A, Stathmin-2, SMEI, and / or any combination(s) thereof.

[0021] These and other embodiments are described in greater detail below.BRIEF DESCRIPTION OF THE DRAWINGS

[0022] FIG. 1A depicts a bar graph of AAV-luciferase formulation stability following five freeze / thaw cycles (5x FT) as measured in a size exclusion chromatography (SEC) assay as a percentage of total peak area of high molecular weight (HMW) species.

[0023] FIG. IB depicts a bar graph of AAV-luciferase formulation stability following five freeze / thaw cycles (5x FT) as measured in a DNA labeling assay as a percentage change compared to levels of DNA labeled at the zero time point (TO).

[0024] FIG. 2A depicts bar graphs of AAV-luciferase formulation stability following exposure to oxidation as measured in a size exclusion chromatography (SEC) assay as a percentage of total peak area of high molecular weight (HMW) species.

[0025] FIG. 2B depicts a bar graph of AAV-luciferase formulation stability following five freeze / thaw cycles (5x FT) as measured in a DNA labeling assay as a percentage change compared to levels of DNA labeled at the zero time point (TO).

[0026] FIG. 3A depicts a bar graph of AAV-luciferase formulation stability following exposure to thermal stress (45°C) as measured by recovery of gene of interest (GOI) titer.

[0027] FIG. 3B depicts a bar graph of AAV-luciferase formulation stability following exposure to thermal stress (45°C) as measured in a size exclusion chromatography (SEC) assay as a percentage of total peak area of high molecular weight (HMW) species.

[0028] FIG. 3C depicts a bar graph of AAV-luciferase formulation stability following exposure to thermal stress (45°C) as measured in a DNA labeling assay as a percentage change compared to levels of DNA labeled at the zero time point (TO).DETAILED DESCRIPTION

[0029] Some embodiments herein are directed to the use of one or more amino acids to improve resistance of the formulation to one or more stressors as compared to the same formulation without the amino acids. Some embodiments herein are directed to a formulation, the formulation comprising: (a) a buffer, (b) one or more salts, (c) one or more amino acids, and (d) a detergent. In some embodiments, the detergent is present in an amount ranging between 0.0001% and 1%; and wherein the formulation has a pH ranging between 7 and 9, for example between about 7.5 to about 8.5, about 7.7 to about 8.3, or about 8. In several embodiments, the formulation has a pH of 8.Terms

[0030] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as is commonly understood by one of ordinary skill in the art. All patents, applications, published applications and other publications referenced herein are incorporated by reference in their entirety unless stated otherwise. In the event that there are a plurality of definitions for a term herein, those in this section prevail unless stated otherwise.

[0031] As used herein, and unless otherwise specified, the terms “about” and “approximately,” when used in connection with doses, amounts or weight percents of ingredients of a composition or a dosage form, mean a dose, amount or weight percent that isrecognized by one of ordinary skill in the art to provide a pharmacological effect equivalent to that obtained from the specified dose, amount, or weight percent. In some embodiments, the terms “about” and “approximately,” when used in this context, contemplate a dose, amount, or weight percent within 30%, within 20%, within 15%, within 10%, or within 5%, of the specified dose, amount or weight percent. Where percentages are provided for agents, ingredients and compounds, they can be %m / m, %m / w, %w / w, %m / v, %v / v and variations thereof with respect to the formulation as a whole, unless otherwise indicated.

[0032] Where a range of values is provided, it is understood that the upper and lower limit, and each intervening value between the upper and lower limit of the range is encompassed within the embodiments.

[0033] Terms and phrases used in this application, and variations thereof, especially in the appended claims, unless otherwise expressly stated, should be construed as open ended as opposed to limiting. As examples of the foregoing, the term ‘including’ should be read to mean ‘including, without limitation,’ ‘including but not limited to,’ or the like; the term ‘comprising’ as used herein is synonymous with ‘including,’ ‘containing,’ or ‘characterized by,’ and is inclusive or open-ended and does not exclude additional, unrecited elements or method steps; the term ‘having’ should be interpreted as ‘having at least;’ the term ‘includes’ should be interpreted as ‘includes but is not limited to;’ the term ‘example’ is used to provide exemplary instances of the item in discussion, not an exhaustive or limiting list thereof; and use of terms like ‘preferably,’ ‘preferred,’ ‘desired,’ or ‘desirable,’ and words of similar meaning should not be understood as implying that certain features are critical, essential, or even important to the structure or function, but instead as merely intended to highlight alternative or additional features that may or may not be utilized in a particular embodiment. In addition, the term “comprising” is to be interpreted synonymously with the phrases "having at least" or "including at least". When used in the context of a process, the term "comprising" means that the process includes at least the recited steps but may include additional steps. When used in the context of a compound, composition or device, the term "comprising" means that the compound, composition or device includes at least the recited features or components but may also include additional features or components.

[0034] With respect to the use of substantially any plural and / or singular terms herein, those having skill in the art can translate from the plural to the singular and / or from thesingular to the plural as is appropriate to the context and / or application. The various singular / plural permutations may be expressly set forth herein for sake of clarity. The indefinite article “a” or “an” does not exclude a plurality. The mere fact that certain measures are recited in mutually different dependent claims does not indicate that a combination of these measures cannot be used to advantage. Any reference signs in the claims should not be construed as limiting the scope.

[0035] As used herein, the term “dialysis” has its plain and ordinary meaning as understood in light of the specification and may also refer to the separation of particles in a liquid based on differences in their ability to pass through a membrane.

[0036] As used herein, the term “drug substance” has its plain and ordinary meaning as understood in light of the specification and refers to a therapeutic AAV that has undergone cesium chloride ultracentrifugation purification followed by buffer exchange with a formulation buffer. A drug substance may refer to an AAV that is in a buffer that is suitable for administration to a subject. In such cases, the formulation buffer comprises a suitable salt concentration such that the drug substance may be safely administered to a subject.

[0037] As used herein, the term “forced degradation conditions” has its plain and ordinary meaning as understood in light of the specification and refers to, for example, freeze thaw cycles, exposure to oxidation, and accelerated storage stability conditions.

[0038] As used herein, the term “DNA labeling” has its plain and ordinary meaning as understood in light of the specification and refers to DNA labeling with the reagent, Diamond Dye, to measure levels of viral DNA ejected from capsids following exposure to degradation conditions. When viral DNA is no longer encapsidated by the capsid protein, it complexes with the Diamond Dye reagent in the formulation buffer and fluoresces at approximately 537 nm following excitation at 473 nm. When the capsid is stressed, the viral DNA is often ejected and subsequently exposed to the formulation matrix which includes Diamond Dye. An increase in the fluorescent signal is indicative of capsid breakdown.

[0039] As used herein, the term “ddPCR” has its plain and ordinary meaning as understood in light of the specification and refers to Droplet Digital Polymerase Chain Reaction (ddPCR). It refers to a method of determining the number of encapsidated copies of a viral gene in a sample and is reported as a concentration (vg / mL) where “vg” was vector genomes.

[0040] As used herein, the term “agitation stress” has its plain and ordinary meaning as understood in light of the specification and may also refer to a physical stress that formulations, for example protein therapeutics, are subjected to during routine manufacturing processes. Agitation stress may occur as a result of mixing, ultrafiltration / diafiltration, pumping, shipping, and / or filling.

[0041] As used herein, the term “shear stress” has its plain and ordinary meaning as understood in light of the specification and may also refer to the component of stress that acts parallel to a material cross section. The most common source of shear stress occurs when forces are applied directly parallel to a surface like the fluid shear stress that occurs in vascular tissue from flowing blood interacting with the vessel wall.

[0042] As used herein, the term “freeze-thaw cycle” has its plain and ordinary meaning as understood in light of the specification and may also refer to the lowering of a temperature below the freezing point of the substance followed by the subsequent heating to a temperature above the freezing point of the substance.

[0043] As used herein, the term “thermal stress” has its plain and ordinary meaning as understood in light of the specification and may also refer to mechanical stress created by a change in temperature of a material or inequalities in the temperatures of different portions of the same material.

[0044] As used herein, the term “dynamic light scattering” (DLS) has its plain and ordinary meaning as understood in light of the specification and may also refer to a technique for determining the size distribution profile of small particles in suspension or polymers in solution. In the scope of DLS, temporal fluctuations are usually analyzed using the intensity or photon autocorrelation function.

[0045] The term “AAV” is an abbreviation for adeno-associated virus and may be used to refer to the virus itself or derivatives thereof. The term covers all subtypes and both naturally occurring and recombinant forms, unless otherwise indicated. The abbreviation “rAAV” refers to recombinant adeno-associated virus, also referred to as a recombinant AAV vector (or “rAAV vector”), which refers to AAV comprising a polynucleotide sequence not of AAV origin (e.g., a transgene). The term “AAV” includes AAV1, AAV2, AAV2G9, AAV3, AAV3a, AAV3b, AAV3-3, AAV4, AAV4-4, AAV5, AAV6, AAV6.1, AAV6.2, AAV6.1.2, AAV7, AAV7.2, AAV8, AAV9, AAV9.11, AAV9.13, AAV9.16, AAV9.24, AAV9.45,AAV9.47, AAV9.61, AAV9.68, AAV9.84, AAV9.9, AAV10, AAV11, AAV12. AAV16.3, AAV24.1, AAV27.3, AAV42.12, AAV42-lb, AAV42-2, AAV42-3a, AAV42-3b, AAV42-4, AAV42-5a, AAV42-5b, AAV42-6b, AAV42-8, AAV42-10, AAV42-11, AAV42-12, AAV42-13, AAV42-15, AAV42-aa, AAV43-1, AAV43-12, AAV43-20, AAV43-21, AAV43- 23, AAV43-25, AAV43-5, AAV44.1, AAV44.2, AAV44.5, AAV223.1, AAV223.2, AAV223.4, AAV223.5, AAV223.6, AAV223.7, AAVl-7 / rh.48, AAVl-8 / rh.49, AAV2- 15 / rh.62, AAV2-3 / rh.61, AAV2-4 / rh.5O, AAV2-5 / rh.51, AAV3.1 / hu.6, AAV3.1 / hu.9, AAV3- 9 / rh.52, AAV3-1 l / rh.53, AAV4-8 / rl 1.64, AAV4-9 / rh.54, AAV4-19 / rh.55, AAV5-3 / rh.57, AAV5-22 / rh.58, AAV7.3 / hu.7, AAV16.8 / hu.lO, AAV16.12 / hu. l 1, AAV29.3 / bb.l, AAV29.5 / bb.2, AAV106.1 / hu.37, AAV114.3 / hu.4O, AAV127.2 / hu.41, AAV127.5 / hu.42, AAV128.3 / hu.44, AAV130.4 / hu.48, AAV145.1 / hu.53, AAV145.5 / hu.54, AAV145.6 / hu.55, AAV161.1O / hu.6O, AAV161.6 / hu.61, AAV33.12 / hu.l7, AAV33.4 / hu.l5, AAV33.8 / hu. l6, AAV52 / hu. l9, AAV52.1 / hu.2O, AAV58.2 / hu.25, AAVA3.3, AAVA3.4, AAVA3.5, AAV A3.7, AAVC1, AAVC2, AAVC5, AAV-DJ, AAV-DJ8, AAVF3, AAVF5, AAVH2, AAVrh.72, AAVhu.8, AAVrh.68, AAVrh.70, AAVpi. l, AAVpi.3, AAVpi.2, AAVrh.60, AAVrh.44, AAVrh.65, AAVrh.55, AAVrh.47, AAVrh.69, AAVrh.45, AAVrh.59, AAVhu.12, AAVH6, AAVLKO3, AAVH-l / hu.l, AAVH-5 / hu.3, AAVLG-10 / rh.40, AAVLG-4 / rh.38, AAVLG-9 / hu.39, AAVN721-8 / rh.43, AAVCh.5, AAVCh.5Rl, AAVcy.2, AAVcy.3, AAVcy.4, AAVcy.5, AAVCy.5Rl, AAVCy.5R2, AAVCy.5R3, AAVCy.5R4, AAVcy.6, AAVhu.1, AAVhu.2, AAVhu.3, AAVhu.4, AAVhu.5, AAVhu.6, AAVhu.7, AAVhu.9, AAVhu. lO, AAVhu.11, AAVhu.13, AAVhu.15, AAVhu.16, AAVhu.17, AAVhu.18,AAVhu.20, AAVhu.21, AAVhu.22, AAVhu.23.2, AAVhu.24, AAVhu.25, AAVhu.27,AAVhu.28, AAVhu.29, AAVhu.29R, AAVhu.31, AAVhu.32, AAVhu.34, AAVhu.35,AAVhu.37, AAVhu.39, AAVhu.40, AAVhu.41, AAVhu.42, AAVhu.43, AAVhu.44,AAVhu.44Rl, AAVhu.44R2, AAVhu.44R3, AAVhu.45, AAVhu.46, AAVhu.47, AAVhu.48, AAVhu.48R1, AAVhu.48R2, AAVhu.48R3, AAVhu.49, AAVhu.51, AAVhu.52, AAVhu.54, AAVhu.55, AAVhu.56, AAVhu.57, AAVhu.58, AAVhu.60, AAVhu.61, AAVhu.63, AAVhu.64, AAVhu.66, AAVhu.67, AAVhu.14 / 9, AAVhu.t 19, AAVrh.2, AAVrh.2R. AAVrh.8, AAVrh.8R, AAVrh.10, AAVrh.12, AAVrh.13, AAVrh. l3R, AAVrh.14, AAVrh.17, AAVrh.18, AAVrh.19, AAVrh.20, AAVrh.21, AAVrh.22, AAVrh.23, AAVrh.24, AAVrh.25, AAVrh.31, AAVrh.32, AAVrh.33, AAVrh.34, AAVrh.35, AAVrh.36, AAVrh.37,AAVrh.37R2, AAVrh.38, AAVrh.39, AAVrh.40, AAVrh.46, AAVrh.48, AAVrh.48.1, AAVrh.48.1.2, AAVrh.48.2, AAVrh.49, AAVrb.51, AAVrh.52, AAVrh.53, AAVrh.54, AAVrh.56, AAVrh.57, AAVrh.58, AAVrh.61, AAVrh.64, AAVrh.64Rl, AAVrh.64R2, AAVrh.67, AAVrh.73. AAVrh.74, AAVrh8R, AAVrh8R A586R mutant, AAVrh8R R533A mutant, AAAV, BAAV, caprine AAV, bovine AAV, ovine AAV, AAVhEl.l, AAVhErl.5, AAVhER1.14, AAVhErl.8, AAVhErl.16, AAVhErl.18, AAVhErl.35, AAVhErl.7, AAVhErl.36, AAVhEr2.29, AAVhEr2.4, AAVhEr2.16, AAVhEr2.30, AAVhEr2.31, AAVhEr2.36, AAVhER1.23, AAVhEr3.1, AAV2.5T, AAV-PAEC, AAV-LK01, AAV- LK02, AAV-LK03, AAV-LK04, AAV-LK05, AAV-LK06, AAV-LK07, AAV-LK08, AAV- LK09, AAV-LK10, AAV-LK11, AAV-LK12, AAV-LK13, AAV-LK14, AAV-LK15, AAV- LK16, AAV-LK17, AAV-LK18, AAV-LK19, AAV-PAEC2, AAV-PAEC4, AAV-PAEC6, AAV-PAEC7, AAV-PAEC8, AAV-PAEC11, AAV-PAEC12, AAV-2-pre-miRNA-101, AAV-8h, AAV-8b, AAV-h, AAV-b, AAV SM 10-2, AAV Shuffle 100-1, AAV Shuffle 100- 3, AAV Shuffle 100-7, AAV Shuffle 10-2, AAV Shuffle 10-6, AAV Shuffle 10-8, AAV Shuffle 100-2, AAV SM 10-1, AAV SM 10-8, AAV SM 100-3, AAV SM 100-10, BNP61 AAV, BNP62 AAV, BNP63 AAV, AAVrh.50, AAVrh.43, AAVrh.62, AAVrb.48, AAVhu.19, AAVhu.l l, AAVhu.53, AAV4-8 / rh.64, AAVLG-9 / hu.39, AAV54.5 / hu.23, AAV54.2 / hu.22, AAV54.7 / hu.24, AAV54.1 / hu.21, AAV54.4R / hu.27, AAV46.2 / hu.28, AAV46.6 / hu.29, AAV128.1 / hu.43, true type AAV (ttAAV), UPENN AAV 10, Japanese AAV 10 serotypes, AAV CBr-7.1, AAV CBr-7.10, AAV CBr-7.2, AAV CBr-7.3, AAV CBr-7.4, AAV CBr-7.5, AAV CBr-7.7, AAV CBr-7.8, AAV CBr-B7.3, AAV CBr-B7.4, AAV CBr-El, AAV CBr-E2, AAV CBr-E3, AAV CBr-E4, AAV CBr-E5, AAV CBr-e5, AAV CBr-E6, AAV CBr-E7, AAV CBr-E8, AAV CHt-1, AAV CHt-2, AAV CHt-3, AAV CHt-6.1, AAV CHt-6.10, AAV CHt- 6.5, AAV CHt-6.6, AAV CHt-6.7, AAV CHt-6.8, AAV CHt-Pl, AAV CHt-P2, AAV CHt-P5, AAV CHt-P6, AAV CHt-P8, AAV CHt-P9, AAV CKd-1, AAV CKd-10, AAV CKd-2, AAV CKd-3, AAV CKd-4, AAV CKd-6, AAV CKd-7, AAV CKd-8, AAV CKd-Bl, AAV CKd- B2, AAV CKd-B3, AAV CKd-B4, AAV CKd-B5, AAV CKd-B6, AAV CKd-B7, AAV CKd- B8, AAV CKd-Hl, AAV CKd-H2, AAV CKd-H3, AAV CKd-H4, AAV CKd-H5, AAV CKd- H6, AAV CKd-N3, AAV CKd-N4, AAV CKd-N9, AAV CLg-Fl, AAV CLg-F2, AAV CLg- F3, AAV CLg-F4, AAV CLg-F5, AAV CLg-F6, AAV CLg-F7, AAV CLg-F8, AAV CLv-1, AAV CLv-1, AAV Clvl-10, AAV CLvl-2, AAV CLv-12, AAV CLvl-3, AAV CLv-13, AAVCLvl -4, AAV Civ 1 -7, AAV Civ 1 -8, AAV Civ 1-9, AAV CLv-2, AAV CLv-3, AAV CLv-4, AAV CLv-6, AAV CLv-8, AAV CLv-Dl, AAV CLv-D2, AAV CLv-D3, AAV CLv-D4, AAV CLv-D5, AAV CLv-D6, AAV CLv-D7, AAV CLv-D8, AAV CLv-El, AAV CLv-Kl, AAV CLv-K3, AAV CLv-K6, AAV CLv-L4, AAV CLv-L5, AAV CLv-L6, AAV CLv-Ml, AAV CLv-Ml l, AAV CLv-M2, AAV CLv-M5, AAV CLv-M6, AAV CLv-M7, AAV CLv-M8, AAV CLv-M9, AAV CLv-Rl, AAV CLv-R2, AAV CLv-R3, AAV CLv-R4, AAV CLv-R5, AAV CLv-R6, AAV CLv-R7, AAV CLv-R8, AAV CLv-R9, AAV CSp-1, AAV CSp-10, AAV CSp-11, AAV CSp-2, AAV CSp-3, AAV CSp-4, AAV CSp-6, AAV CSp-7, AAV CSp- 8, AAV CSp-8.10, AAV CSp-8.2, AAV CSp-8.4, AAV CSp-8.5, AAV CSp-8.6, AAV CSp- 8.7, AAV CSp-8.8, AAV CSp-8.9, AAV CSp-9, AAV.hu.48R3, AAV.VR-355, AAV3B, AAV4, AAV5, AAVF1 / HSC1, AAVF11 / HSC11, AAVF12 / HSC12, AAVF13 / HSC13, AAVF14 / HSC14, AAVF15 / HSC15, AAVF16 / HSC16, AAVF17 / HSC17, AAVF2 / HSC2, AAVF3 / HSC3, AAVF4 / HSC4, AAVF5 / HSC5, AAVF6 / HSC6, AAVF7 / HSC7, AAVF8 / HSC8, AAVF9 / HSC9, PHP.B, PHP. A, G2B-26, G2B-13, THE 1-32, THE 1-35, rAAV, myotropic AAV variants, AAV-SLB101, AAV-SLB101 variants, MyoAAV and AAVMYO, AAV-DJ, AAV-DJ8, or variants thereof.

[0046] The term “AAV virus” or “AAV viral particle” or “rAAV vector particle” refers to a viral particle composed of at least AAV capsid protein and an encapsidated polynucleotide.

[0047] The term “AAV-SLB101” refers to a myotropic viral construct comprising a modified AAV9 having an RGDLGLS (SEQ ID NO: 1) peptide inserted between positions 588 and 589 of VPl.

[0048] The term “adeno-associated virus (AAV) capsid” refers to the three- dimensional proteinaceous shell encapsidating, including, for example enclosing, the viral genetic material. The AAV capsid is a non-enveloped, icosahedral 60-mer of three repeating monomers: VP1, VP2, and VP3. The AAV capsid determines the properties of viral particles, including tissue tropism and antigenic properties.

[0049] The term “tropism” refers to preferential entry of the AAV virus or viral particle into certain cell or tissue type(s) and / or preferential interaction with the cell surface that facilitates entry into certain cell or tissue types, optionally and preferably followed by expression (e.g., transcription and, optionally, translation) of one or more nucleic acidsequences (e g., one or more transgene sequences) carried by the AAV virus or viral particle. Variations in capsid proteins VP1, VP2, and / or VP3, or a combination thereof may alter AAV virus tropism.

[0050] The terms “variant adeno-associated virus (AAV) capsid” and “engineered adeno-associated virus (AAV) capsid” refer to an AAV capsid polypeptide sequence differing by at least one amino acid from a parent, or non-engineered, polypeptide sequence. Variant or engineered AAV capsid may also refer to an AAV capsid comprising one or more engineered features.

[0051] The term “engineered feature” refers to alterations in the AAV capsid polypeptide at specific loci such that the one or more viral properties, for example, transduction or tropism of the AAV virus or viral particle, is altered.

[0052] The terms “polypeptide” and “protein” are used interchangeably to refer to a polymer of amino acid residues and are not limited to a minimum length. Polypeptides, including therapeutic proteins and other peptides, e.g., linkers, tags, capsid proteins, may include amino acid residues including natural and / or non-natural amino acid residues. The terms also include post-expression modifications of the polypeptide, for example, glycosylation, sialylation, acetylation, phosphorylation, and the like. In some respects, the polypeptides may contain modifications with respect to a native or natural sequence, if the protein maintains the desired activity. These modifications may be deliberate, as through site- directed mutagenesis, or may be accidental, such as through mutations of hosts which produce the proteins or errors due to PCR amplification.

[0053] Amino acids generally can be grouped according to the following common side- chain properties: (1) hydrophobic: Norleucine, Met, Ala, Vai, Leu, He; (2) neutral hydrophilic: Cys, Ser, Thr, Asn, Gin; (3) acidic: Asp, Glu; (4) basic: His, Lys, Arg; (5) residues that influence chain orientation: Gly, Pro; (6) aromatic: Trp, Tyr, Phe.

[0054] Amino acid substitutions may also refer to one or more changes in a polypeptide sequence. The changes may include replacement of one amino acid in a polypeptide with another amino acid, insertion of one or amino acids, and / or deletion of one or more amino acids, or any combination thereof. Non-conservative amino acid substitutions will involve exchanging a member of one of these classes for another class.

[0055] A “nucleic acid” sequence refers to a deoxyribonucleic acid (DNA) or ribonucleic acid (RNA) sequence. The term captures sequences that include any of the known base analogues of DNA and RNA such as, but not limited to 4-acetylcytosine, 8-hydroxy-N6- methyladenosine, aziridinylcytosine, pseudoisocytosine, 5-(carboxyhydroxyl- methyl) uracil, 5 -fluorouracil, 5-bromouracil, 5- carboxymethylaminomethyl-2-thiouracil, 5- carboxymethylaminomethyluracil, dihydrouracil, inosine, N6-isopentenyladenine, 1- methyladenine, 1- methylpseudouracil, 1 -methylguanine, 1 -methylinosine, 2,2- dimethylguanine, 2-m ethyladenine, 2-methylguanine, 3 -methylcytosine, 5-methylcytosine, N6-methyladenine, 7-methylguanine, 5-methylaminomethyluracil, 5-methoxy- aminomethyl- 2-thiouracil, beta-D-mannosylqueosine, 5'-methoxycarbonylmethyluracil, 5-methoxyuracil, 2- methylthio-N6-isopentenyladenine, uracil-5-oxyacetic acid methylester, uracil-5-oxyacetic acid, oxybutoxosine, pseudouracil, queosine, 2-thiocytosine, 5- methyl-2-thiouracil, 2- thiouracil, 4-thiouracil, 5-methyluracil, N- uracil-5-oxyacetic acid methylester, uracil-5- oxyacetic acid, pseudouracil, queosine, 2-thiocytosine, and 2,6-diaminopurine.

[0056] The term “polynucleotide,” refers to a polymeric form of nucleotides of any length, including DNA, RNA, or analogs thereof. A polynucleotide may comprise modified nucleotides, such as methylated nucleotides and nucleotide analogs, and may be interrupted by non-nucleotide components. If present, modifications to the nucleotide structure may be imparted before or after assembly of the polymer. The term polynucleotide, as used herein, refers interchangeably to double- and single-stranded molecules. Unless otherwise specified or required, any embodiment of the invention described herein that is a polynucleotide encompasses both the double-stranded form and each of two complementary single-stranded forms known or predicted to make up the double-stranded form.

[0057] The term “polymerase chain reaction” has its plain and ordinary meaning, including but not limited to referring to a technique for copying or amplifying a nucleic acid sequence. Quantitative (qPCR) couples amplification of a target nucleic acid sequence with quantification of the concentration of the nucleic acid sequence in a sample. Droplet digital polymerase chain reaction (ddPCR) is a third generation of polymerase chain reaction (PCR) that enables the exact quantification of nucleic acid targets within a sample.

[0058] The term “isolated” when referring to a nucleotide sequence has its plain and ordinary meaning, including but not limited to meaning that the indicated molecule ispresent in the substantial absence of other biological macromolecules of the same type. Thus, an “isolated nucleic acid molecule which encodes a particular polypeptide” refers to a nucleic acid molecule which is substantially free of other nucleic acid molecules that do not encode the subject polypeptide; however, the molecule may include some additional bases or moieties which do not materially affect the basic characteristics of the composition.

[0059] The term “recombinant,” as applied to a polynucleotide has its plain and ordinary meaning, including but not limited to meaning that the polynucleotide is the product of various combinations of cloning, restriction or ligation steps, and other procedures that result in a construct that is distinct from a polynucleotide found in nature and / or a combination of polynucleotides and viral proteins that is not found in nature. A recombinant virus is a viral particle comprising a recombinant polynucleotide. The terms respectively include replicates of the original polynucleotide construct and progeny of the original virus construct.

[0060] The term “gene,” has its plain and ordinary meaning, including but not limited to referring to a polynucleotide containing at least one open reading frame that can encode a particular gene product. Any of the polynucleotide sequences described herein may be used to identify larger fragments or full-length coding sequences of the genes with which they are associated. Methods of isolating larger fragment sequences are known to those of skill in the art.

[0061] The term “transgene,” as used herein, has its plain and ordinary meaning, including but not limited to referring to a nucleic acid sequence to be positioned within a viral vector and encoding a polypeptide, protein or other product of interest. In some embodiments, one rAAV vector, or engineered rAAV vector may comprise a sequence encoding one or more transgenes (which can optionally be the same gene, or different genes). For example, one rAAV vector may comprise the coding sequence for 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 transgenes.

[0062] The terms “gene transfer” or “gene delivery” refer to methods or systems for inserting DNA, such as a transgene, into host cells, such as those of a subject afflicted with cardiomyopathy. In several embodiments, gene transfer yields transient expression of nonintegrated transferred DNA, extrachromosomal replication and expression of transferred replicons (e.g., episomes). In additional embodiments, gene transfer results in integration of transferred genetic material into the genomic DNA of host cells.

[0063] Gene therapy drug products (such as rAAV particles) are challenging to incorporate into composition and formulations due to their limited stability in the liquid state and a propensity for large-scale aggregation at low concentrations. Gene therapy drug products are often delivered directly to treatment areas (including CNS tissue); which requires that excipients and formulation parameters be compatible with tissue function, microenvironment, and volume restrictions.

[0064] In some embodiments disclosed herein, AAV particles may be prepared as, or included in, pharmaceutical compositions. Such compositions may include one or more active ingredients and, most often, one or more pharmaceutically acceptable excipients.

[0065] Relative amounts of the active ingredient (e.g. AAV particle), a pharmaceutically acceptable excipient, and / or any additional ingredients in a pharmaceutical composition in accordance with the present disclosure may vary, depending upon the identity, size, and / or condition of the subject being treated and further depending upon the route by which the composition is to be administered. For example, the composition may include between 0.1% and 99% (w / w) of the active ingredient. By way of example, the composition may include between 0.1% and 100%. e.g., between 0.5 and 50%, between 1-30%, between 5- 80%, or at least 80% (w / w) active ingredient.

[0066] Although the descriptions of pharmaceutical compositions provided herein are principally directed to pharmaceutical compositions which are suitable for administration to humans, it will be understood by the skilled artisan that such compositions are generally suitable for administration to any other animal, e.g., to non-human animals, e g., non-human mammals. Modification of pharmaceutical compositions suitable for administration to humans in order to render the compositions suitable for administration to various animals is well understood, and the ordinarily skilled veterinary pharmacologist can design and / or perform such modification with merely ordinary, if any, experimentation. Subjects to which administration of the pharmaceutical compositions is contemplated include, but are not limited to, humans and / or other primates; mammals, including commercially relevant mammals such as cattle, pigs, horses, sheep, cats, dogs, mice, rats, birds, including commercially relevant birds such as poultry, chickens, ducks, geese, and / or turkeys.

[0067] In some embodiments, the formulations may comprise at least one inactive ingredient. As used herein, the term “inactive ingredient” refers to one or more inactive agentsincluded in formulations. In some embodiments, all, none or some of the inactive ingredients which may be used in the formulations of the present disclosure may be approved by the US Food and Drug Administration (FDA). Formulations comprising amino acids for intravenous administration which may be used in the formulations of the present disclosure have not been approved by the US Food and Drug Administration (FDA).

[0068] As used herein, the term “water for injection” (WFI) has its plain and ordinary meaning and may refer to water that is of extra high quality without significant contamination. In some embodiments, sterile WFI may used for making preparing the formulations disclosed herein. In some embodiments, WFI is used in the preparation of a formulation disclosed herein for administration by injection. Before such use other substances, for example, but not limited to NaCl, may be added to make the solution isotonic. Isotonic solutions containing water for injection can be given by injection into a vein, muscle, or under the skin. A non-sterile version may be used in manufacturing with sterilization occurring later in the production process.

[0069] Formulations and pharmaceutical compositions described herein may be prepared by any method known or hereafter developed in the art of pharmacology. In general, such preparatory methods include the step of bringing the active ingredient into association with an excipient and / or one or more other accessory ingredients, and then, if necessary and / or desirable, dividing, shaping and / or packaging the product into a desired single- or multi-dose unit.

[0070] A pharmaceutical composition in accordance with the present disclosure may be prepared, packaged, and / or sold in bulk, as a single unit dose, and / or as a plurality of single unit doses. As used herein, a “unit dose” refers to a discrete amount of the pharmaceutical composition comprising a predetermined amount of the active ingredient. The amount of the active ingredient is generally equal to the dosage of the active ingredient which would be administered to a subject and / or a convenient fraction of such a dosage such as, for example, one-half or one-third of such a dosage.

[0071] According to the present disclosure AAV particles may be formulated for CNS delivery. Agents that cross the brain blood barrier may be used. For example, some cell penetrating peptides that can target molecules to the brain blood barrier endothelium may beused for formulation (e.g., Mathupala, Expert Opin Ther Pat., 2009, 19, 137-140; the content of which is incorporated herein by reference in its entirety).

[0072] Formulations of the present disclosure can be used in any step of producing, processing, preparing, storing, expanding, or administering AAV particles and viral vectors. In certain embodiments, pharmaceutical formulations and components can be use in AAV production, AAV processing, AAV clarification, AAV purification, and AAV finishing systems of the present disclosure, all of which can be pre-rinsed, packed, equilibrated, flushed, processed, eluted, washed, or cleaned with formulations known to those in the art, including AAV pharmaceutical, processing and storage formulations of the present disclosure.

[0073] The terms “regulatory element” or “regulatory sequence”, or variations thereof, refer to a nucleotide sequence that participates in functional regulation of a polynucleotide, including replication, duplication, transcription, splicing, translation, or degradation of the polynucleotide. Regulatory elements can be enhancing or inhibitory depending on the embodiment. Non-limiting examples of regulatory elements include transcriptional regulatory sequences such as promoter sequences, polyadenylation signals, transcription termination sequences, upstream regulatory domains, origins of replication, internal ribosome entry sites (“IRES”), enhancers, and the like. These elements collectively provide for the replication, transcription, and translation of a coding sequence in a recipient cell, though not all these sequences need always be present. It shall be appreciated that the structural components of a rAAV vector as provided for herein may be listed in individual paragraphs solely for clarity and may be used together in combination. For example, any regulatory element or other component can be used in combination with any transgene (or transgenes) provided for herein.

[0074] The term “vector,” as used herein, refers to a nucleic acid molecule capable of propagating another nucleic acid to which it is linked. The term includes the vector as a selfreplicating nucleic acid structure as well as the vector incorporated into the genome of a host cell into which it has been introduced. Certain vectors are capable of directing the expression of nucleic acids to which they are operatively linked. Such vectors are referred to herein as “expression vectors.”

[0075] An “expression vector” is a vector comprising a region of nucleic acid (e.g., a transgene) which encodes a gene product (e.g., a polypeptide or protein) of interest. Asdisclosed herein, vectors are used for achieving expression, e.g., stable expression, of a protein in an intended target cell. An expression vector may also comprise control elements operatively linked to the transgene to facilitate expression of the encoded protein in the target cell. A combination of one or more regulatory elements and a gene or genes to which they are operably linked for expression may be referred to herein as an “expression cassette.”

[0076] As used herein, the term “reduced capillary isoelectric focusing” (R-cIEF) has its plain and ordinary meaning as understood in light of the specification and may also refer to an electrophoretic technique to separate sample components based on isoelectric point (pl). The relative purity of the main species of the sample was reported in relation to more acidic (lower pl) and more basic (higher pl) species.

[0077] As used herein, the term “reduced capillary electrophoresis” (R-CE) has its plain and ordinary meaning as understood in light of the specification. This is an electrophoretic technique that allows for separation of the three viral capsid proteins (VP1, VP2, and VP3). The relative total purity is determined based on peak area counts. For these studies, the change in the distribution of the acidic, main, and basic species is evaluated. This change is typically seen as an increase in acidic species with a corresponding decrease in the main following stress.

[0078] As used herein, the term “isoelectric point” (pl) has its plain and ordinary meaning as understood in light of the specification and may also refer to the pH value at which a molecule carries no electrical charge.

[0079] As used herein, the term “Size Exclusion Chromatography” (SEC) has its plain and ordinary meaning as understood in light of the specification and may also refer to a chromatographic technique that separated sample components based on molecular size and shape. The relative total purity of the monomeric species is determined in relation to other sized components identified as either high molecular weight (HMW) or low molecular weight (LMW) species. For these studies, the change in the distribution of monomer, HMW, and LMW is evaluated. As the sample is stressed, there is typically an increase in HMW content. The unstressed sample (TO) was considered nominal. A formulation with a higher percentage of HMW species was considered less desirable than one with a lower % HMW species.

[0080] A “subject” refers to mammal that is the object of treatment using a method or composition as provided for herein. “Mammal” includes, without limitation, mice, rats,rabbits, guinea pigs, dogs, cats, sheep, goats, cows, horses, primates, such as monkeys, chimpanzees, and apes, and humans. In some embodiments, the subject is human.

[0081] As used herein, the terms “treat,” “treating,” “treatment,” “therapeutic,” and “therapy” do not necessarily mean total cure or abolition of the disease or condition. Any alleviation of any undesired signs or symptoms of the disease or condition, to any extent can be considered treatment and / or therapy. Furthermore, treatment may include acts that may worsen the subject’s overall feeling of well-being or appearance.

[0082] The term “effective amount” is used to indicate an amount of an active compound, or pharmaceutical agent, that elicits the biological or medicinal response indicated. For example, an effective amount of compound, salt or composition can be the amount needed to prevent, alleviate or ameliorate symptoms of the disease or condition, or prolong the survival of the subject being treated. This response may occur in a tissue, system, animal or human and includes alleviation of the signs or symptoms of the disease or condition being treated. Determination of an effective amount is well within the capability of those skilled in the art, in view of the disclosure provided herein. The effective amount of the compounds disclosed herein required as a dose will depend on the route of administration, the type of animal, including human, being treated, and the physical characteristics of the specific animal under consideration. The dose can be tailored to achieve a desired effect, but will depend on such factors as weight, diet, concurrent medication and other factors which those skilled in the medical arts will recognize.

[0083] As used herein, a “composition” refers to any mixture of two or more products, substances, or compounds, including cells. It may be a formulation, solution, a suspension, liquid, powder, a paste, aqueous, non-aqueous or any combination thereof.

[0084] As used herein, the term “kit” may be used to describe variations of the portable, self-contained enclosure that includes at least one set of components to conduct one or more of the diagnostic or therapeutic methods of the invention.

[0085] The term “package insert” is used to refer to instructions customarily included in commercial packages of therapeutic products, that contain information about the indications, usage, dosage, administration, combination therapy, contraindications and / or warnings concerning the use of such therapeutic products. Non-limiting Embodiments

[0086] Some embodiments herein relate to the use of one or more excipients, for example, but not limited to, amino acids, salts, base buffers, detergents, and, optionally, stabilizers, in a formulation to promote recovery of a sample from the formulation following one or more stressors, for example, but not limited to, agitation stress, shear stress, freeze-thaw stress, thermal stress, or any combination therein, as compared to the same formulation without the one or more excipients. In some embodiments, the one or more excipients in the formulation promote recovery of about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 20%, 25%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% more of a sample from the formulation buffer following from agitation stress, shear stress, freeze-thaw stress, thermal stress, or any combination therein, as compared to the same formulation without the one or more excipients.

[0087] As described herein, the inclusion of one or more amino acids can assist increasing resistance of a sample to one or more stressors, for example, but not limited to, agitation stress, shear stress, freeze-thaw stress, thermal stress, or any combination therein, as compared to the same formulation without the one or more amino acids. Some embodiments herein relate to the use of one or more amino acids in a formulation to promote recovery from and / or resistance to one or more stressors, for example, but not limited to, agitation stress, shear stress, freeze-thaw stress, thermal stress, or any combination therein, as compared to the same formulation buffer without the one or more amino acids. In some embodiments, the one or more amino acids in the formulation promote recovery of about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 20%, 25%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% more of a sample from the formulation buffer following from agitation stress, shear stress, freeze-thaw stress, thermal stress, or any combination therein, or an amount of sample that is in a range defined by any two of the preceding values.

[0088] In some embodiments, the one or more amino acids in the formulation promote increased sample resistance to freeze-thaw (FT) cycling, as compared to the same formulation buffer without the one or more amino acids. In some embodiments, the one or more amino acids in the formulation promote increased resistance of a sample in the formulation to 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 freeze-thaw cycles, or increased resistance to a number of FT cycles that is in a range defined by any two of the preceding values, as compared to the same formulation without the one or more amino acids. In some embodiments, theformulation promotes recovery of about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 20%, 25%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% more of a sample from the formulation buffer following 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 FT cycles, or an amount of sample that is in a range defined by any two of the preceding values.

[0089] In some embodiments, the one or more amino acids in the formulation promote increased sample resistance to agitation, as compared to the same formulation without the one or more amino acids. In some embodiments, the one or more amino acids in the formulation promote recovery of about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 20%, 25%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% more of a sample from the formulation buffer following agitation at about 1 rpm, 2 rpm, 3 rpm, 4 rpm, 5 rpm, 6 rpm, 7 rpm, 8 rpm, 9 rpm, 10 rpm, 20 rpm, 25 rpm, 30 rpm, 40 rpm, 50 rpm, 60 rpm, 70 rpm, 75 rpm, 80 rpm, 90 rpm, 100 rpm, 125 rpm, 150 rpm, 175 rpm, 200 rpm, 250 rpm, 300 rpm, 400 rpm, or 500 rpm, as compared to the same formulation buffer without the one or more amino acids.

[0090] In some embodiments, the one or more amino acids in the formulation buffer promote increased sample resistance to thermal stress as compared to the same formulation without the one or more amino acids. In some embodiments, the one or more amino acids in the formulation promote recovery of about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 20%, 25%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% more of a sample from the formulation following exposure to temperatures of about 1°C, 2°C, 3°C, 4°C, 5°C, 6°C, 7°C, 8°C, 9°C, 10°C, 20°C, 25°C, 30°C, 40°C, 45°C, 50°C, 55°C, or 60°C, as compared to the same formulation without the one or more amino acids.

[0091] In some embodiments, the formulations disclosed herein comprise one or more amino acids. In some embodiments, the amino acid wherein the one or more amino acids increases a stress resistance of the formulation as compared to the stress resistance of the same formulation in the absence of the one or more amino acids. For example, in some embodiments, the one or more amino acids increases resistance to agitation stress, shear stress, freeze-thaw stress, thermal stress, or any combination therein. In some embodiments, the one or more amino acids comprise alanine, arginine, glutamic acid, glycine, methionine, proline, and / orserine. In some embodiments, the formulation comprises about 0.1 mM, 0.5 mM, 1 mM, 2 mM, 3 mM, 4 mM, 5 mM, 6 mM, 7 mM, 8 mM, 9 mM, 10 mM, 15 mM, 20 mM, 25 mM, 30 mM, 40 mM, 50 mM, 60 mM, 70 mM, 75 mM, 80 mM, 90 mM, 100 mM, 125 mM, 250 mM, 175 mM, or 200 mM alanine, arginine, glutamic acid, glycine, methionine, proline, and / or serine, or an amount of alanine, arginine, glutamic acid, glycine, methionine, proline, and / or serine that is in a range defined by any two of the preceding values. For example, in some embodiments, the formulation comprises between about 0.1-200 mM, 0.1-150 mM, 0.1-100 mM, 0.1-75 mM, 0.1-50 mM, 0.1-25 mM, 0.1-10 mM, 0.1-1 mM, 1-200 mM, 1-250 mM, 1- 100 mM, 1-75 mM, 1-50 mM, 1-25 mM, 1-10 mM, 10-200 mM, 10-150 mM, 10-100 mM, 10- 75 mM, 10-50 mM, 10-25 mM, 25-200 mM, 25-150 mM, 25-100 mM, 25-75 mM, 25-50 mM, 50-200 mM, 50-150 mM, 50-100 mM, 100-200 mM, 100-150 mM, or 150-200 mM, alanine, arginine, glutamic acid, glycine, methionine, proline, and / or serine. In some embodiments, the one or more amino acids comprise arginine. In some embodiments, the formulation comprises about 20 mM, 25 mM, 30 mM, 40 mM, 50 mM, 60 mM, or 70 mM, arginine, or an amount of arginine that is in a range defined by any two of the preceding values. For example, in some embodiments, the formulation comprises between about 20-70 mM, 20-60 mM, 20-50 mM, 20-40 mM, 20-30 mM, 30-70 mM, 30-60 mM, 30-50 mM, 30-40 mM, 40-70 mM, 40-60 mM, 40-50 mM, 50-70 mM, 50-60 mM, or 60-70 mM arginine. In some embodiments, the one or more amino acids comprise proline. In some embodiments, the formulation comprises about 20 mM, 25 mM, 30 mM, 40 mM, 50 mM, 60 mM, or 70 mM arginine, or an amount of proline that is in a range defined by any two of the preceding values. For example, in some embodiments, the formulation comprises between about 20-70 mM, 20-60 mM, 20-50 mM, 20-40 mM, 20-30 mM, 30-70 mM, 30-60 mM, 30-50 mM, 30-40 mM, 40-70 mM, 40-60 mM, 40-50 mM, 50-70 mM, 50-60 mM, or 60-70 mM proline. In some embodiments, the one or more amino acids comprises arginine and proline. In some embodiments, the formulation comprises about 20 mM, 25 mM, 30 mM, 40 mM, 50 mM, 60 mM, or 70 mM, arginine, or an amount of arginine that is in a range defined by any two of the preceding values and about 20 mM, 25 mM, 30 mM, 40 mM, 50 mM, 60 mM, or 70 mM, proline, or an amount of proline that is in a range defined by any two of the preceding values. For example, in some embodiments, the formulation comprises between about 20-70 mM, 20-60 mM, 20-50 mM, 20-40 mM, 20-30 mM, 30-70 mM, 30-60 mM, 30-50 mM, 30-40 mM, 40-70 mM, 40-60 mM,40-50 mM, 50-70 mM, 50-60 mM, or 60-70 mM arginine and about 20-70 mM, 20-60 mM, 20-50 mM, 20-40 mM, 20-30 mM, 30-70 mM, 30-60 mM, 30-50 mM, 30-40 mM, 40-70 mM, 40-60 mM, 40-50 mM, 50-70 mM, 50-60 mM, or 60-70 mM proline.

[0092] In some embodiments, the formulation comprises one or more base buffers. In some embodiments, the one or more base buffers comprise acetate, citrate, histidine, phosphate, succinate, Tris, and / or Tris HCL In some embodiments, the formulation comprises about 5 mM, 6 mM, 7 mM, 8 mM, 9 mM, 10 mM, 11 mM, 12 mM, 13 mM, 14 mM, 15 mM, 16 mM, 17 mM, 18 mM, 19 mM, 20 mM, 21 mM, 22 mM, 23 mM, 24 mM, or 25 mM, of one or more of acetate, citrate, histidine, phosphate, succinate, and / or Tris, or an amount of acetate, citrate, histidine, phosphate, succinate, Tris, and / or Tris HC1, that is in a range defined by any of the preceding values. In some embodiments, the one or more base buffers comprises Tris. In some embodiments, the formulation comprises about 5 mM, 6 mM, 7 mM, 8 mM, 9 mM, 10 mM, 11 mM, 12 mM, 13 mM, 14 mM, 15 mM, 16 mM, 17 mM, 18 mM, 19 mM, 20 mM, 21 mM, 22 mM, 23 mM, 24 mM, or 25 mM, Tris and / or Tris HC1 or an amount of Tris and / or Tris HC1 that is in a range defined by any of the preceding values. In some embodiments, the formulation comprises about 5 mM, 6 mM, 7 mM, 8 mM, 9 mM, 10 mM, 11 mM, 12 mM, 13 mM, 14 mM, 15 mM, 16 mM, 17 mM, 18 mM, 19 mM, 20 mM, 21 mM, 22 mM, 23 mM, 24 mM, or 25 mM, or an amount of Tris or Tris HC1, that is in a range defined by any of the preceding values. In some embodiments, the formulation comprises about 5 mM, 6 mM, 7 mM, 8 mM, 9 mM, 10 mM, 11 mM, 12 mM, 13 mM, 14 mM, 15 mM, 16 mM, 17 mM, 18 mM, 19 mM, 20 mM, 21 mM, 22 mM, 23 mM, 24 mM, or 25 mM, tris and Tris-HCl in combination or an amount of tris and tris HCL, that is in a range defined by any of the preceding values. In several embodiments, the buffer comprises about 20 mM Tris. In several embodiments, the buffer comprises 20 mM Tris. In several embodiments, the buffer comprises about 20 mM of Tris and Tris-HCl in combination. In several embodiments, the buffer comprises 20 mM of Tris and Tris-HCl in combination.

[0093] In some embodiments, the formulation comprises one or more salts. In some embodiments, the one or more salts comprises MgCk and / or NaCl. In some embodiments, the formulation comprises about 0.1 mM, 0.5 mM, 1 mM, 5 mM, 10 mM, 15 mM, 20 mM, 25 mM, 30 mM, 40 mM, 50 mM, 57 mM, 60 mM, 70 mM, 75 mM, 80 mM, 90 mM, 100 mM, 125 mM, 150 mM, 175 mM, 200 mM, or 250mM MgC12 and / or NaCl, or an amount of MgCkand / or NaCl that is in a range defined by any two of the preceding values. In some embodiments, the one or more salts comprise MgCh. In some embodiments, the formulation comprises about 0.1 mM, 0.2 mM, 0.3 mM, 0.4 mM, 0.5 mM, 0.6 mM, 0.7 mM, 0.8 mM, 0.9 mM, 1 mM, 2 mM, 3 mM, 4 mM, or 5 mM MgCk, or an amount of MgCh that is in a range defined by any two of the preceding values. For example, in some embodiments, the formulation comprises between about 0.1-5 mM, 0.1-3 mM, 0.1-1 mM, 0.1-0.5 mM, 0.5-5 mM, 0.5-3 mM, 0.5-1 mM, 1-5 mM, 1-3 mM, or 3-5 mM MgCk. In some embodiments, the formulation comprises about 0.1 mM, 0.5 mM, 1 mM, 5 mM, 10 mM, 15 mM, 20 mM, 25 mM, 30 mM, 40 mM, 50 mM, 57 mM, 60 mM, 70 mM, 75 mM, 80 mM, 90 mM, 100 mM, 110 mM, 120 mM, 125 mM, 130 mM, 140 mM, 150 mM, 160 mM, 170 mM, 175 mM, 180 mM, 190 mM, 200 mM, or 250 mM NaCl, or an amount of NaCl that is in a range defined by any two of the preceding values. In some embodiments, the one or more salts in the formulation promote recovery of a sample from the formulation following one or more stressors, for example, but not limited to, agitation stress, shear stress, freeze-thaw stress, thermal stress, or any combination therein. In some embodiments, the formulation exhibits improved stability by the addition of sodium chloride under forced degradation conditions due to storage at accelerated temperatures and multiple freeze-thaw cycles.

[0094] In some embodiments, the formulation comprises one or more detergents. In some embodiments, the formulation comprises one or more non-ionic detergents. In some embodiments, the formulation comprises one or more ionic, anionic, and / or cationic detergents. In some embodiments, the formulation comprises one or more polysorbates, for example, but not limited to, polysorbate 20, 40, 60, 65, and / or 80. In some embodiments, the formulation comprises one or more sorbitans, for example, but not limited to sorbitan stearate, sorbitan laurate, sorbitan sesquioleate, sorbitan oleate, sorbitan Tristearate, sorbitan palmitate and / or sorbitan trioleate. In some embodiments, the formulation comprises one or more poloxamers, for example, but not limited to, poloxamer 68, 88, 98, 108, 124, 188, 237, 338, and 407. In some embodiments, the detergent comprises sodium lauryl sulfate. In some embodiments, the formulation comprises benzalkonium chloride. In some embodiments, the formulation comprises centrimonium bromide. In some embodiments, the formulation comprises cocamidopropyl betaine. In some embodiments, the formulation comprises sodium cocoamphoacetate. In some embodiments, the one or more detergents comprise Pl 88 and / orPS80. In some embodiments, the formulation comprises about 0.001%, 0.002%, 0.003%, 0.004%, 0.005%, 0.006%, 0.007%, 0.008%, 0.009%, 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09%, or 0.1 %, P188 and / or polysorbate 80 (PS80), or an amount of P188 and / or PS80 that is in a range defined by any two of the preceding values.

[0095] In some embodiments, the formulation optionally comprises one or more stabilizers. In some embodiments, the one or more stabilizers comprise HSA, cyclodextrin, sucrose, and / or sorbitol. In some embodiments, the formulation comprises about 0.00%, 0.01%, 0.05%, 0.1%, 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, or 20%, HSA, cyclodextrin, sucrose, and / or sorbitol, or an amount of HSA, cyclodextrin, sucrose, and / or sorbitol that is in a range defined by any two of the preceding values. In some embodiments, the formulation comprises about 1 mM, 2 mM, 3 mM, 4 mM, 5 mM, 6 mM, 7 mM, 8 mM, 9 mM, 10 mM, 20 mM, 30 mM, 40 mM, 50 mM, 60 mM, 70 mM, 80 mM, 90 mM, 100 mM, 110 mM, 120 mM, 130 mM, 140 mM, 150 mM, 160 mM, 170 mM, 180 mM, 190 mM, 200 mM, 210 mM, 220 mM, 230 mM, 240 mM, 250 mM, 260 mM, 270 mM, 280 mM, 290 mM, or 300 mM, HSA, cyclodextrin, sucrose, and / or sorbitol, or an amount of HSA, cyclodextrin, sucrose, and / or sorbitol that is in a range defined by any two of the preceding values.

[0096] In some embodiments, the pH of the formulation is maintained. In some embodiments, the formulation comprises a pH ranging between about 7 and 9. In some embodiments, the formulation comprises a pH of about 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8.0, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, 9.0, or a pH that is in a range defined by any two of the preceding values. In several embodiments, the formulation has a pH of between 7.5 and 8.5. In several embodiments, the formulation has a pH of between 7.7 and 8.3. In several embodiments, the formulation has a pH of about 8. In several embodiments, the formulation has a pH of 8.

[0097] Where numerical values are provided herein, including the tables below, those values may be for individual ingredients or for groups of ingredients. In some embodiments, the formulation comprises, consists essentially of or consists of several (e.g., 3 or 4) or all of the following groups of ingredients listed in Table 1.Table 1 - Formulations

[0098] Some embodiments herein are directed to a formulation, the formulation comprising: (a) a buffer, (b) one or more salts, (c) one or more amino acids, and (d) a detergent. In some embodiments, the buffer comprises acetate, citrate, histidine, phosphate, succinate, and / or Tris, or any combination thereof. In some embodiments, the one or more salts comprise MgCh and / or NaCl. In some embodiments, the one or more amino acids comprise alanine, arginine, glutamic acid, glycine, methionine, proline, and / or serine. In some embodiments, the one or more detergents comprise Pl 88 and / or PS80. In some embodiments, the formulation further comprises one or more agents that act as a stabilizer. In some embodiments, the one or more stabilizing agents comprise HSA, cyclodextrin, sucrose, and / or sorbitol. In some embodiments, the formulation further comprises a therapeutic protein or molecule. In someembodiments, the formulation comprises a drug substance. Tn some embodiments, the drug substance comprises a therapeutic molecule. In some embodiments, the therapeutic molecule comprises a nucleic acid. In some embodiments, the formulation comprises a vector. In some embodiments, the vector is an adenovirus associated viral (AAV) vector. In some embodiments, the formulation is formulated for therapeutic use. In some embodiments, the formulation must be further prepared for therapeutic use. In some embodiments, preparation for therapeutic use may comprise adjusting the concentration of the drug substance, and or one or more parameters of the formulation, for example, but not limited to, the pH and / or osmolality of the formulation.

[0099] Some embodiments herein are directed to a formulation, the formulation comprising: (a) 20 mM Tris, (b) 1 mM MgCh, (c) 50 mM arginine, (d) 50 mM proline, and (e) 0.005% P188, wherein the formulation is at a pH of about 8. In some embodiments, the formulation has an osmolality of 155-202 mOsm / kg. In some embodiments, the formulation further comprises WFI. Some embodiments herein are directed to a formulation, the formulation comprising: (a) about 5-25 mM Tris / Tris-HCl in combination, (b) about 0.1-5 mM MgCh (c) about 100-300 mM NaCl, (d) about 20-70 mM, (e) about 20-70 mM, and (f) about 0.001-0.01% P188, wherein the formulation is at a pH of about 8. In some embodiments, the formulation has an osmolality of about 150-250 mOsm / kg. In some embodiments, the formulation further comprises WFI. Some embodiments herein are directed to a formulation, the formulation comprising: (a) 5-25 mM Tris, (b) 0.1-5 mM MgCh (c) 50-300 mM NaCl, (d) 20-70 mM, (e) 20-70 mM, and (f) 0.001-0.01% P188, wherein the formulation is at a pH of about 8. In some embodiments, the formulation has an osmolality of 150-250 mOsm / kg. In some embodiments, the formulation further comprises WFI. Some embodiments herein are directed to a formulation, the formulation comprising: (a) 9-12 mM Tris, (b) 0.1-5 mM MgCh (c) 50-300 mM NaCl, (d) 20-70 mM, (e) 20-70 mM, and (f) 0.001-0.01% P188, wherein the formulation is at a pH of about 8. In some embodiments, the formulation has an osmolality of 150-250 mOsm / kg. In some embodiments, the formulation further comprises WFI.

[0100] Some embodiments herein are directed to a formulation, the formulation comprising: (a) 20 mM Tris / Tris HC1, (b) 1 mM MgCh, (c) 50 mM arginine, (d) 50 mM proline, (e) 75 mM NaCl, and (f) 0.005% P188, wherein the formulation is at a pH of about 8. In some embodiments, the formulation has an osmolality of 280-340 mOsm / kg. In someembodiments, the formulation further comprises WFI. Some embodiments herein are directed to a formulation, the formulation comprising: (a) about 5-25 mM Tris, (b) about 0.1-5 mM MgCh, (c) about 20-70 mM arginine, (d) about 20-70 mM proline, (e) about 50-100 mM NaCl, and (f) about 0.005% P188, wherein the formulation is at a pH of about 8. In some embodiments, the formulation has an osmolality of about 280-340 mOsm / kg. In some embodiments, the formulation further comprises WFI. Some embodiments herein are directed to a formulation, the formulation comprising: (a) 5-25 mM Tris, (b) 0.1-5 mM MgCk, (c) 20- 70 mM arginine, (d) 20-70 mM proline, (e) 50-100 mM NaCl, and (f) 0.005% Pl 88, wherein the formulation is at a pH of about 8. In some embodiments, the formulation has an osmolality of 280-340 mOsm / kg. In some embodiments, the formulation further comprises WFI. Some embodiments herein are directed to a formulation, the formulation comprising: (a) 9-12 mM Tris / Tris-HCl in combination, (b) 0.1-5 mM MgCh, (c) 20-70 mM arginine, (d) 20-70 mM proline, (e) 50-100 mM NaCl, and (f) 0.005% Pl 88, wherein the formulation is at a pH of about 8. In some embodiments, the formulation has an osmolality of 280-340 mOsm / kg. In some embodiments, the formulation further comprises WFI. In some embodiments, the formulation meets the requirements for formulation of an isotonic drug product. In some embodiments, the formulation comprises a pH suitable for intravenous administration.

[0101] Formulations disclosed herein may be hypotonic compared to physiological levels and thus may require dilution, for example, but not limited to, dilution in a hypertonic saline solution prior to administration.

[0102] In some embodiments, the formulation is provided in a ready to administer form. In some embodiments, the ready to administer formulation comprises a single-dose, sterile, preservative-free suspension for intravenous infusion. In some embodiments, the formulation is supplied in a 10 mb single-use cyclic olefin polymer (COP) vial with a 9.0 mb deliverable volume. In some embodiments, the drug product contains a target concentration of 2.0E13vg / mL viral vector in a formulation of 20 mM Tris buffer containing 1 mM magnesium chloride, 50 mM L proline, 50 mM L arginine, and 0.005% poloxamer 188 at pH 8.0. In some embodiments, the drug product contains a target concentration of 2.0E13vg / mL viral vector in a formulation of 20 mM Tris / Tris-HCl buffer containing 1 mM magnesium chloride, 50 mM L proline, 50 mM L arginine, 75mM NaCl, and 0.005% poloxamer 188 at pH 8.0. In some embodiments, the drug product contains a target concentration of 2.0E13vg / mL viral vector ina formulation of 10 mM Tris buffer containing 1 mM magnesium chloride, 50 mM L proline, 50 mM L arginine, and 0.005% poloxamer 188 at pH 8.0. In some embodiments, the drug product contains a target concentration of 2.0E13vg / mL viral vector in a formulation of 10 mM Tris / Tris-HCl buffer containing 1 mM magnesium chloride, 50 mM L proline, 50 mM L arginine, 75mM NaCl, and 0.005% poloxamer 188 at pH 8.0. In some embodiments, the formulation exhibits improved stability by the addition of sodium chloride under forced degradation conditions due to storage at accelerated temperatures and multiple freeze-thaw cycles.

[0103] In some embodiments, the formulation is provided in a form that must be further prepared before administration. In some embodiments, the formulation is provided in multiple vials, for example in 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 different vials. In some embodiments, thr formulations is provided in 6 vials. In some embodiments, each of the vials are placed at room temperature and allowed to fully thaw. In some embodiments, the vials are placed at room temperature for 0.1, 0.25, 0.5, 0.75, 1, 2, 2.5, 3, 3.5, 4, 4.5, or 5 hours, of for a time in a range defined by any two of the preceding values. In some embodiments, preparation of the formulation comprises mixing the contents of the vial, for example, but not limited to gentle mixing by inversion. Following mixing the contents of the vial may be pooled into a single vessel. The pooled formulation is then diluted, for example, but not limited to, by adding 1 part 3% saline to 6 parts of the pooled formulation. The clinical formulation may then be transferred into an IV bag.

[0104] In some embodiments, the formulation buffer further comprises a drug product. In some embodiments, the drug product is a therapeutic drug product. In some embodiments, the drug product is a biological sample. In some embodiments, the drug product comprises a nucleic acid, polypeptide, protein, or any combination thereof. In some embodiments, the drug product comprises an expression cassette. In some embodiments, the expression cassette is composed of, at a minimum, a transgene and its regulatory sequences, including Kozak sequences. Where the cassette is designed to be expressed from a rAAV, the expression cassette further contains 5' and 3' AAV ITRs. These ITR's may be full-length, or one or both of the ITRs may be truncated. In some embodiments, the rAAV is pseudotyped, for example, the AAV capsid is from a different source AAV than that the AAV which provides the ITRs. In some embodiments, the serotype of the AAV may comprise AAV1,AAV2, AAV2G9, AAV3, AAV3a, AAV3b, AAV3-3, AAV4, AAV4-4, AAV5, AAV6, AAV6.1, AAV6.2, AAV6.1.2, AAV7, AAV7.2, AAV8, AAV9, AAV9.11, AAV9.13, AAV9.16, AAV9.24, AAV9.45, AAV9.47, AAV9.61, AAV9.68, AAV9.84, AAV9.9, AAV10, AAV11, AAV12. AAV16.3, AAV24.1, AAV27.3, AAV42.12, AAV42-lb, AAV42- 2, AAV42-3a, AAV42-3b, AAV42-4, AAV42-5a, AAV42-5b, AAV42-6b, AAV42-8, AAV42-10, AAV42-11, AAV42-12, AAV42-13, AAV42-15, AAV42-aa, AAV43-1, AAV43- 12, AAV43-20, AAV43-21, AAV43-23, AAV43-25, AAV43-5, AAV44.1, AAV44.2, AAV44.5, AAV223.1, AAV223.2, AAV223.4, AAV223.5, AAV223.6, AAV223.7, AAV1- 7 / rh.48, AAVl-8 / rh.49, AAV2-15 / rh.62, AAV2-3 / rh.61, AAV2-4 / rh.5O, AAV2-5 / rh.51, AAV3.1 / hu.6, AAV3.1 / hu.9, AAV3-9 / rh.52, AAV3-1 l / rh.53, AAV4-8 / rl 1.64, AAV4- 9 / rh.54, AAV4-19 / rh.55, AAV5-3 / rh.57, AAV5-22 / rh.58, AAV7.3 / hu.7, AAV16.8 / hu.lO, AAV16.12 / hu.l l, AAV29.3 / bb.l, AAV29.5 / bb.2, AAV106.1 / hu.37, AAV114.3 / hu.4O, AAV127.2 / hu.41, AAV127.5 / hu.42, AAV128.3 / hu.44, AAV130.4 / hu.48, AAV145.1 / hu.53, AAV145.5 / hu.54, AAV145.6 / hu.55, AAV161.1O / hu.6O, AAV161.6 / hu.61, AAV33.12 / hu.l7, AAV33.4 / hu.l5, AAV33.8 / hu.l6, AAV52 / hu. l9, AAV52.1 / hu.2O, AAV58.2 / hu.25, AAV A3.3, AAV A3.4, AAV A3.5, AAV A3.7, AAVC1, AAVC2, AAVC5, AAV-DJ, AAV- DJ8, AAVF3, AAVF5, AAVH2, AAVrh.72, AAVhu.8, AAVrh.68, AAVrh.70, AAVpi.l, AAVpi.3, AAVpi.2, AAVrh.60, AAVrh.44, AAVrh.65, AAVrh.55, AAVrh.47, AAVrh.69, AAVrh.45, AAVrh.59, AAVhu.12, AAVH6, AAVLKO3, AAVH-l / hu. l, AAVH-5 / hu.3, AAVLG-10 / rh.40, AAVLG-4 / rh.38, AAVLG-9 / hu.39, AAVN721-8 / rh.43, AAVCh.5, AAVCh.5Rl, AAVcy.2, AAVcy.3, AAVcy.4, AAVcy.5, AAVCy.5Rl, AAVCy.5R2, AAVCy.5R3, AAVCy.5R4, AAVcy.6, AAVhu.l, AAVhu.2, AAVhu.3, AAVhu.4, AAVhu.5, AAVhu.6, AAVhu.7, AAVhu.9, AAVhu.lO, AAVhu. l l, AAVhu.13, AAVhu.15, AAVhu.16, AAVhu.17, AAVhu.18, AAVhu.20, AAVhu.21, AAVhu.22, AAVhu.23.2, AAVhu.24, AAVhu.25, AAVhu.27, AAVhu.28, AAVhu.29, AAVhu.29R, AAVhu.31, AAVhu.32, AAVhu.34, AAVhu.35, AAVhu.37, AAVhu.39, AAVhu.40, AAVhu.41, AAVhu.42, AAVhu.43, AAVhu.44, AAVhu.44Rl, AAVhu.44R2, AAVhu.44R3, AAVhu.45, AAVhu.46, AAVhu.47, AAVhu.48, AAVhu.48Rl, AAVhu.48R2, AAVhu.48R3, AAVhu.49, AAVhu.51, AAVhu.52, AAVhu.54, AAVhu.55, AAVhu.56, AAVhu.57, AAVhu.58, AAVhu.60, AAVhu.61, AAVhu.63, AAVhu.64, AAVhu.66, AAVhu.67, AAVhu.14 / 9, AAVhu.t 19, AAVrh.2, AAVrh.2R. AAVrh.8, AAVrh.8R, AAVrh. lO, AAVrh.12, AAVrh.13, AAVrh. l3R,AAVrh.14, AAVrh.17, AAVrh.18, AAVrh.19, AAVrh.20, AAVrh.21, AAVrh.22, AAVrh.23, AAVrh.24, AAVrh.25, AAVrh.31, AAVrh.32, AAVrh.33, AAVrh.34, AAVrh.35, AAVrh.36, AAVrh.37, AAVrh.37R2, AAVrh.38, AAVrh.39, AAVrh.40, AAVrh.46, AAVrh.48, AAVrh.48.1, AAVrh.48.1.2, AAVrh.48.2, AAVrh.49, AAVrb.51, AAVrh.52, AAVrh.53, AAVrh.54, AAVrh.56, AAVrh.57, AAVrh.58, AAVrh.61, AAVrh.64, AAVrh.64Rl, AAVrh.64R2, AAVrh.67, AAVrh.73. AAVrh.74, AAVrh8R, AAVrh8R A586R mutant, AAVrh8R R533A mutant, AAAV, BAAV, caprine AAV, bovine AAV, ovine AAV, AAVhEl.l, AAVhErl.5, AAVhER1.14, AAVhErl.8, AAVhErl.16, AAVhErl.18, AAVhErl.35, AAVhErl.7, AAVhErl.36, AAVhEr2.29, AAVhEr2.4, AAVhEr2.16, AAVhEr2.30, AAVhEr2.31, AAVhEr2.36, AAVhER1.23, AAVhEr3.1, AAV2.5T, AAV- PAEC, AAV-LK01, AAV-LK02, AAV-LK03, AAV-LK04, AAV-LK05, AAV-LK06, AAV- LK07, AAV-LK08, AAV-LK09, AAV-LK10, AAV-LK11, AAV-LK12, AAV-LK13, AAV- LK14, AAV-LK15, AAV-LK16, AAV-LK17, AAV-LK18, AAV-LK19, AAV-PAEC2, AAV-PAEC4, AAV-PAEC6, AAV-PAEC7, AAV-PAEC8, AAV-PAEC11, AAV-PAEC12, AAV-2-pre-miRNA-101, AAV-8h, AAV-8b, AAV-h, AAV-b, AAV SM 10-2, AAV Shuffle 100-1, AAV Shuffle 100-3, AAV Shuffle 100-7, AAV Shuffle 10-2, AAV Shuffle 10-6, AAV Shuffle 10-8, AAV Shuffle 100-2, AAV SM 10-1, AAV SM 10-8, AAV SM 100-3, AAV SM 100-10, BNP61 AAV, BNP62 AAV, BNP63 AAV, AAVrh.50, AAVrh.43, AAVrh.62, AAVrb.48, AAVhu.19, AAVhu.l l, AAVhu.53, AAV4-8 / rh.64, AAVLG-9 / hu.39, AAV54.5 / hu.23, AAV54.2 / hu.22, AAV54.7 / hu.24, AAV54.1 / hu.21, AAV54.4R / hu.27, AAV46.2 / hu.28, AAV46.6 / hu.29, AAV128.1 / hu.43, true type AAV (ttAAV), UPENN AAV 10, Japanese AAV 10 serotypes, AAV CBr-7.1, AAV CBr-7.10, AAV CBr-7.2, AAV CBr- 7.3, AAV CBr-7.4, AAV CBr-7.5, AAV CBr-7.7, AAV CBr-7.8, AAV CBr-B7.3, AAV CBr- B7.4, AAV CBr-El, AAV CBr-E2, AAV CBr-E3, AAV CBr-E4, AAV CBr-E5, AAV CBr- e5, AAV CBr-E6, AAV CBr-E7, AAV CBr-E8, AAV CHt-1, AAV CHt-2, AAV CHt-3, AAV CHt-6.1, AAV CHt-6.10, AAV CHt-6.5, AAV CHt-6.6, AAV CHt-6.7, AAV CHt-6.8, AAV CHt-Pl, AAV CHt-P2, AAV CHt-P5, AAV CHt-P6, AAV CHt-P8, AAV CHt-P9, AAV CKd- 1, AAV CKd-10, AAV CKd-2, AAV CKd-3, AAV CKd-4, AAV CKd-6, AAV CKd-7, AAV CKd-8, AAV CKd-Bl, AAV CKd-B2, AAV CKd-B3, AAV CKd-B4, AAV CKd-B5, AAV CKd-B6, AAV CKd-B7, AAV CKd-B8, AAV CKd-Hl, AAV CKd-H2, AAV CKd-H3, AAV CKd-H4, AAV CKd-H5, AAV CKd-H6, AAV CKd-N3, AAV CKd-N4, AAV CKd-N9, AAVCLg-Fl, AAV CLg-F2, AAV CLg-F3, AAV CLg-F4, AAV CLg-F5, AAV CLg-F6, AAV CLg-F7, AAV CLg-F8, AAV CLv-1, AAV CLv-1, AAV Clvl-10, AAV CLvl-2, AAV CLv- 12, AAV CLvl-3, AAV CLv-13, AAV CLvl-4, AAV Civ 1-7, AAV Civ 1-8, AAV Civ 1-9, AAV CLv-2, AAV CLv-3, AAV CLv-4, AAV CLv-6, AAV CLv-8, AAV CLv-Dl, AAV CLv-D2, AAV CLv-D3, AAV CLv-D4, AAV CLv-D5, AAV CLv-D6, AAV CLv-D7, AAV CLv-D8, AAV CLv-El, AAV CLv-Kl, AAV CLv-K3, AAV CLv-K6, AAV CLv-L4, AAV CLv-L5, AAV CLv-L6, AAV CLv-Ml, AAV CLv-Ml l, AAV CLv-M2, AAV CLv-M5, AAV CLv-M6, AAV CLv-M7, AAV CLv-M8, AAV CLv-M9, AAV CLv-Rl, AAV CLv-R2, AAV CLv-R3, AAV CLv-R4, AAV CLv-R5, AAV CLv-R6, AAV CLv-R7, AAV CLv-R8, AAV CLv-R9, AAV CSp-1, AAV CSp-10, AAV CSp-11, AAV CSp-2, AAV CSp-3, AAV CSp-4, AAV CSp-6, AAV CSp-7, AAV CSp-8, AAV CSp-8.10, AAV CSp-8.2, AAV CSp- 8.4, AAV CSp-8.5, AAV CSp-8.6, AAV CSp-8.7, AAV CSp-8.8, AAV CSp-8.9, AAV CSp- 9, AAV.hu.48R3, AAV.VR-355, AAV3B, AAV4, AAV5, AAVF1 / HSC1, AAVF11 / HSC11, AAVF12 / HSC12, AAVF13 / HSC13, AAVF14 / HSC14, AAVF15 / HSC15, AAVF16 / HSC16, AAVF17 / HSC17, AAVF2 / HSC2, AAVF3 / HSC3, AAVF4 / HSC4, AAVF5 / HSC5, AAVF6 / HSC6, AAVF7 / HSC7, AAVF8 / HSC8, AAVF9 / HSC9, PHP.B, PHP.A, G2B-26, G2B-13, TH1.1-32, TH1.1-35, rAAV, myotropic AAV variants, AAV-SLB101, AAV- SLB101 variants, MyoAAV and AAVMYO, AAV-DJ, AAV-DJ8, or any of the modified serotypes of the present disclosure, or variants thereof. In additional embodiments, the ITRs of AAV serotype 1 are used. However, ITRs from other suitable sources may be selected.

[0105] In some embodiments, the concentration of AAV particle in the formulation may be between about l>< 1012VG / ml and about l * 1016VG / ml. As used herein, “VG / ml” represents vector genomes (VG) per milliliter (ml). VG / ml also may describe genome copy per milliliter or DNase resistant particle per milliliter. In some embodiments, the concentration of AAV particle in the formulation is between IxlO12and 5><1013, between 1 x 1012and 5x 1012, between 2xl012and I x lO13, between 5x 1012and I xlO13, between N IO13and 2x l013, between 2x l013and 3x l013, between 2x l013and 2.5 x lO13, between 2.5 x lO13and 3xl013, or no more than 5x l013VG / ml. In some embodiments, the concentration of AAV particle in the formulation is 2.7-3.5x 1013VG / ml. In certain embodiments, the concentration of AAV particle in the formulation is 7. CL IO13VG / ml. In certain embodiments, the concentration of AAV particle in the formulation is 5.0x l012VG / mL. In certain embodiments, the concentration ofAAV particle in the formulation may be between about l * 106total capsid / mL and about 1 x 1016total capsid / ml.

[0106] In some embodiments, the formulation comprises an expression cassette. In some embodiments, the expression cassette is a therapeutic expression cassette. In some embodiments, the therapeutic expression cassette encodes a therapeutic protein. In some embodiments, the therapeutic protein may comprise, for example, frataxin, GSD1 A, PRKAG2, BAG3, Dysferlin, MicroDys, MYBPC-3, Danon / Lamp2, TMEM43, PKP2, LMNA, TNNT2, RBM20, MYH7, RYR2, CAMKII, PRKAG2, CASQ2, DM1, GRN, GBA, MSH3, UNC13A, Stathmin-2, SMEI, and / or any combination(s) thereof. In some embodiments, the recombinant AAV vector is a recombinant AAV vector of serotype rh74. In some embodiments, the recombinant AAV vector is not a recombinant AAV vector of serotype rh74. In some embodiments, the recombinant AAV vector is a recombinant AAV vector of serotype 9. In some embodiments, the recombinant AAV vector is not a recombinant AAV vector of serotype 9. In some embodiments, the AAV packages one or more transgenes. In some embodiments, the one or more transgenes comprise frataxin, GSD1A, PRKAG2, BAG3, Dysferlin, MicroDys, MYBPC-3, Danon / Lamp2, TMEM43, PKP2, LMNA, TNNT2, RBM20, MYH7, RYR2, CAMKII, PRKAG2, CASQ2, DM1, GRN, GBA, MSH3, UNC13A, Stathmin-2, SMEI, and / or any combination(s) thereof.

[0107] Injectable preparations, for example, sterile injectable aqueous or oleaginous suspensions may be formulated according to the known art using suitable dispersing agents, wetting agents, and / or suspending agents. Sterile injectable preparations may be sterile injectable solutions, suspensions, and / or emulsions in nontoxic parenterally acceptable diluents and / or solvents, for example, as a solution in 1,3 -butanediol. Among the acceptable vehicles and solvents that may be employed are water, Ringer's solution, U.S.P., and isotonic sodium chloride solution. Sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose, any bland fixed oil can be employed including synthetic mono- or di glycerides. Fatty acids such as oleic acid can be used in the preparation of injectables.

[0108] Injectable formulations may be sterilized, for example, by filtration through a bacterial-retaining filter, and / or by incorporating sterilizing agents in the form of sterile solidcompositions which can be dissolved or dispersed in sterile water or other sterile injectable medium prior to use.

[0109] In order to prolong the effect of active ingredients, reduction in the rate of the absorption of active ingredients from subcutaneous or intramuscular injections is achieved. This may be accomplished by the use of liquid suspensions of crystalline or amorphous material with poor water solubility. The rate of absorption of active ingredients depends upon the rate of dissolution which, in turn, may depend upon crystal size and crystalline form. Alternatively, delayed absorption of a parenterally administered drug form is accomplished by dissolving or suspending the drug in an oil vehicle. Injectable depot forms are made by forming microcapsule matrices of the drug in biodegradable polymers such as polylactidepolyglycolide. Depending upon the ratio of drug to polymer and the nature of the particular polymer employed, the rate of drug release can be controlled. Non-limiting examples of other biodegradable polymers include poly(orthoesters) and poly (anhydrides). Depot injectable formulations are prepared by entrapping the drug in liposomes or microemulsions which are compatible with body tissues.

[0110] In some embodiments, formulations disclosed herein are administered as a solo therapeutic or combination therapeutic. In some embodiments, the formulation is administered as a solo therapy. In other embodiments, the formulation is administered as part of a combination therapy. The combination therapy may be in combination with one or more agents, for example, but not limited to small molecule compounds, immunosuppressants, growth factors, small molecule compounds which are antioxidants, anti-inflammatory agents, anti-apoptosis agents, calcium regulators, antiglutamatergic agents, steroids, structural protein inhibitors, compounds involved in muscle function, and compounds involved in metal ion regulation and / or hormones. The combination therapy agents may be administered simultaneously. The combination therapy agents may be administered separately.[01U] In some embodiments, the formulations disclosed herein are suitable for intrathecal administration. The formulations may comprise one or more inactive ingredient and / or one or more additional active ingredient. In some embodiments, the formulations disclosed herein are suitable for intrathecal administration in a mammalian subject, e.g., a human, using components and techniques known in the art.

[0112] Formulations disclosed herein may be sterilized using sterilization techniques, or may be sterile fdtered. The resulting aqueous solutions may be packaged for use as is, or lyophilized, the lyophilized preparation being combined with a sterile solution (to achieve a desired concentration) prior to administration.

[0113] The U.S. Food and Drug Administration (FDA) and other agencies stress the importance of impurity control and reproducibility of compounds in drug development. The process is rigorous and helps ensure that an approved drug works correctly and has health benefits that outweigh its known risks. Based on the data provided herein, formulations that includes one or more amino acids, such as those described herein, are unexpectedly superior for at least the reasons provided herein.

[0114] A formulation as described elsewhere herein, can be administered to such subjects by a variety of methods. In any of the uses or methods described herein, administration can be by various routes known to those skilled in the art, including without limitation intravenous, intramuscular, subcutaneous, systemic, intrathecal, intramyocardial and / or intraperitoneal administration to a subject in need thereof. The amount of a formulation described herein, required for use in treatment will vary not only with the particular therapeutic agent and amino acids included, but also with the route of administration, the nature and / or symptoms of the disease or condition being treated and the age and condition of the patient and will be ultimately at the discretion of the attendant physician or clinician. As will be understood by those of skill in the art, in certain situations it may be necessary to administer the compounds disclosed herein in amounts that exceed, or even far exceed, the dosage ranges described herein in order to effectively and aggressively treat particularly aggressive diseases or conditions.

[0115] The desired dose may conveniently be presented in a single dose or as divided doses administered at appropriate intervals, for example, as two, three, four or more sub-doses per day. The sub-dose itself may be further divided, e.g., into a number of discrete loosely spaced administrations.

[0116] As will be readily apparent to one skilled in the art, the useful in vivo dosage to be administered and the particular mode of administration can vary depending upon the age, weight, the severity of the affliction, and mammalian species treated, the particular compounds employed, and the specific use for which these compounds are employed. The determinationof effective dosage levels, that is the dosage levels necessary to achieve the desired result, can be accomplished by one skilled in the art using routine methods, for example, human clinical trials, in vivo studies and in vitro studies. For example, useful dosages of a formulation disclosed herein, can be determined by comparing their in vitro activity, and in vivo activity in animal models. Such comparison can be done by comparison against an established drug, such as for example another drug that is designed to treat the same or related disease or symptom.

[0117] It should be noted that the attending physician would know how to and when to terminate, interrupt, or adjust administration due to toxicity or organ dysfunctions. Conversely, the attending physician would also know to adjust treatment to higher levels if the clinical response were not adequate (precluding toxicity). The magnitude of an administrated dose in the management of the disorder of interest will vary with the severity of the disease or condition to be treated and to the route of administration. The severity of the disease or condition may, for example, be evaluated, in part, by standard prognostic evaluation methods. Further, the dose and perhaps dose frequency, will also vary according to the age, body weight, and response of the individual patient. A program comparable to that discussed above may be used in veterinary medicine.

[0118] Compounds and formulations disclosed herein can be evaluated for efficacy and toxicity using known methods. For example, the toxicology of a particular compound, or of a subset of the compounds, sharing certain chemical moieties, may be established by determining in vitro toxicity towards a cell line, such as a mammalian, and preferably human, cell line. The results of such studies are often predictive of toxicity in animals, such as mammals, or more specifically, humans. Alternatively, the toxicity of particular compounds in an animal model, such as mice, rats, rabbits, dogs or monkeys, may be determined using known methods. The efficacy of a particular compound may be established using several recognized methods, such as in vitro methods, animal models, or human clinical trials. When selecting a model to determine efficacy, the skilled artisan can be guided by the state of the art to choose an appropriate model, dose, route of administration and / or regime.

[0119] A formulation disclosed herein may be manufactured in a manner that is itself known, e.g., by means of conventional mixing, dissolving, granulating, levigating, emulsifying, encapsulating, or entrapping processes. Additionally, the active ingredients are contained in an amount effective to achieve its intended purpose.

[0120] Multiple techniques of administering a formulation described herein exist in the art including, but not limited to pulmonary, aerosol, injection, infusion and parenteral delivery, including intramuscular, subcutaneous, intravenous, intramedullary injections, intramyocardial, intrathecal, direct intraventricular, intraperitoneal, intranasal and intraocular injections.

[0121] One may also administer a formulation described herein in a local rather than systemic manner, for example, via injection or implantation directly into the affected area, often in a depot or sustained release formulation. Furthermore, one may administer a formulation described herein in a targeted drug delivery system, for example, in a liposome coated with a tissue-specific antibody. The liposomes will be targeted to and taken up selectively by the organ. For example, intranasal or pulmonary delivery to target a respiratory disease or condition may be desirable.

[0122] The formulations may, if desired, be presented in a kit which may contain one or more unit dosage forms containing the active ingredient. The kit may be accompanied by instructions for administration. The kit may also be accompanied with a notice associated with the container in form prescribed by a governmental agency regulating the manufacture, use, or sale of pharmaceuticals, which notice is reflective of approval by the agency of the form of the drug for human or veterinary administration. Such notice, for example, may be the labeling approved by the U.S. Food and Drug Administration for prescription drugs, or the approved product insert. In some embodiments, the kit contains one or more vials of a viral vector pharmaceutical formulation. In some embodiments, each vial contains the viral vector pharmaceutical composition at a dose (e.g., a unit dose) of up to or at about 6.0 x io13vg. In some embodiments, each vial of a viral vector (e.g., each unit dose) of the kit contains the pharmaceutical composition at a dose of about 6.0 x io13vg. In some embodiments, each vial of a viral vector (e.g., each unit dose) of the kit contains the pharmaceutical composition at a dose of up to or at about 1.2 x io14vg. In some embodiments, each vial of a viral vector (e.g., each unit dose) of the kit contains the pharmaceutical composition at a dose of about 1.2 x 1014vg. In some embodiments, each vial of a viral vector (e.g., each unit dose) of the kit contains the pharmaceutical composition at a dose of up to or at about 2.4 x io14vg. In some embodiments, each vial of a viral vector (e.g., each unit dose) of the kit contains the pharmaceutical composition at a dose of about 2.4 x 1014vg. In some embodiments, the viralvector pharmaceutical composition is at a concentration of about 0.1 -5.0 x 1013vg / ml. In some embodiments, each vial contains a single dose of rAAV viral vector. In some embodiments, each vial contains more than a single dose of rAAV viral vector. In some embodiments, each vial contains less than a single dose of rAAV viral vector.

[0123] Dosages of AAV may range from about 1 x io11vg / kg, about 1 x io12vg / kg, about 1 x 1013vg / kg, about 1 x io14vg / kg, about 1 x io15vg / kg, about 1 x 1Q16vg / kg, or more vector genomes per kilogram body weight in an adult or neonate.

[0124] In some embodiments, the AAV is administered at a dose of 1.0 x io12vg - 9.9 x 1014vg. In some embodiments, the AAV is administered at a dose of 5.0 x 1013vg - 3.0 x 1014vg. In some embodiments, the AAV is administered at a dose of up to 6.0 x io13vg. In some embodiments, the AAV is administered at a dose of about 6.0 x io13vg. In some embodiments, the AAV is administered at a dose of up to 1.2 x 1014vg. In some embodiments, the AAV is administered at a dose of about 1.2 x io14vg. In some embodiments, the AAV is administered at a dose of up to 2.4 x io14vg. In some embodiments, the AAV is administered at a dose of about 2.4 x 1014vg.

[0125] In some embodiments, the AAV is administered in a unit dose of about 1.0 x 1013vg - 9.9 x 1014vg. In some embodiments, the AAV is administered in a unit dose of about 1.0 x 1013vg - 5.0 x 1014vg. In some embodiments, the AAV is administered in a unit dose of about 5.0 x io13vg - 3.0 x 1014 vg.

[0126] In some embodiments, the AAV is administered in a unit dose of about 6.0 x 1013vg. In some embodiments, the AAV is administered in a unit dose of about 1.2 x io14vg. some embodiments, the AAV is administered in a unit dose of about 2.4 x io14vg.

[0127] Formulation of AAV viral vector to be administered may vary depending, for example, on the method of administration, the dose volume, and the pharmaceutical excipient. In some embodiments, the rAAV viral vector may be formulated in a sterile isotonic drug solution. In some embodiments, the rAAV viral vector may be formulated in saline solution. In some embodiments, the rAAV viral vector may be formulated in an artificial CSF, e.g., Elliott's B solution. In some embodiments, therapeutic formulation is filtered before administration.

[0128] Some embodiments provided herein are described in the following enumerated alternatives.

[0129] 1 . A formulation, the formulation comprising: (a) 5 to 25 mM Tris, (b) 0.5-1.5 mM MgC12, (c) 25-75 mM arginine, (d) 25-75 mM proline, and (e) 0.001-0.2% P188, wherein the formulation is at a pH of 7.7-8.3.

[0130] 2 A formulation, the formulation comprising: (a) 5 to 25 mM Tris, (b) 0.5-1.5 mM MgC12, (c) 50-250 mM NaCl, (d) 25-75 mM arginine, (e) 25-75 mM proline, and (f) 0.001-0.2% P188, wherein the formulation is at a pH of 7.7-8.3.

[0131] 3 A formulation, the formulation comprising: a buffer, wherein the buffer is present in an amount ranging between 5 mM and 30 mM; one or more salts, wherein the one or more salts are individually present in an amount ranging between 0.1 mM and 200 mM; one or more amino acids, wherein the one or more amino acids are individually present in an amount ranging between 1 and 200 mM; and a detergent, wherein the detergent is present in an amount ranging between 0.0001% and 1%; and wherein the formulation is at a pH ranging between 7.5 and 8.5.

[0132] 4. The formulation of alternative 3, wherein the buffer is selected from the group comprising: acetate, citrate, histidine, phosphate, succinate, Tris, and Tris-HCl, or any combination thereof.

[0133] 5 The formulation of alternative 3 or 4, wherein the buffer comprises Tris or Tris HCL.

[0134] 6 The formulation of any one of alternatives 3-5, wherein the buffer comprises 5 to 25 mM Tris or Tris and Tris HCL in combination.

[0135] 7 The formulation of any one of alternatives 3-5, wherein the buffer comprises: 20 mM Tris, 10 mM Tris, 20 mM Tris and Tris-HCl in combination, or 10 mM Tris and Tris HCL in combination.

[0136] 8 The formulation of any one of alternatives 3-7, wherein the one or more salt comprises MgCh, NaCl, or a combination thereof.

[0137] 9. The formulation of any one of alternatives 3-8, wherein the one or more salts comprises between 0.1 and 5 mM MgCh.

[0138] 10. The formulation of any one of alternatives 3-8, wherein the one or more salt comprises between 50 and 250 mM NaCl.

[0139] 11. The formulation of alternative 8, wherein the one or more salt comprises1 mM MgCh and 75 mM NaCl.

[0140] 12. The formulation of alternative 8, wherein the one or more salt comprises1 mM MgCh and 200 mM NaCl.

[0141] 13. The formulation of any one of alternatives 3-12, wherein the one or more amino acids are selected from the group comprising: alanine, arginine, glutamic acid, glycine, methionine, proline, serine, or any combination thereof.

[0142] 14. The formulation of any one of alternatives 3-13, wherein the one or more amino acids comprises between 25 and 75 mM arginine.

[0143] 15. The formulation of any one of alternatives 3-13, wherein the one or more amino acids comprises between 25 and 75 mM proline.

[0144] 16. The formulation of any one of alternatives 3-13, wherein the one or more amino acids comprises between 25 and 75 mM arginine and between 25 and 75 mM proline.

[0145] 17. The formulation of any one of alternatives 3-16, wherein the detergent comprises Pl 88, PS80, or a combination thereof.

[0146] 18. The formulation of any one of alternatives 3-16, wherein the detergent comprises between 0.001 and 0.01% P188.

[0147] 19. The formulation of any one of alternatives 3-18, wherein the formulation is at a pH ranging between 7.7 and 8.3.

[0148] 20. The formulation of any one of alternatives 3-19, wherein the formulation is at a pH of 8.

[0149] 21. The formulation of alternative 3, wherein the buffer comprises 5 to 25 mM Tris or Tris HC1, the one or more salts comprises 0.5-1.5 mM MgCh, the one or more amino acids comprises 25-75 mM arginine and 25-75 mM proline, and the detergent comprises 0.001-0.2% P188, wherein the formulation is at a pH of 7.7-8.3.

[0150] 22. The formulation of alternative 3, wherein the buffer comprises 5 to 30 mM Tris or Tris HC1, the one or more salts comprises 0.5- 1.5 mM MgCh and 50-250 mM NaCl, the one or more amino acids comprises 30-60 mM arginine and 30-60 mM proline, and the detergent comprises 0.001-0.2% P188, wherein the formulation is at a pH of 7.7-8.3.

[0151] 23. The formulation of any one of the preceding alternatives, wherein the one or more amino acids increases a stress resistance of the formulation as compared to the stress resistance of the same formulation in the absence of the one or more amino acids.

[0152] 24. The formulation of alternative 23, wherein the stress resistance is selected from agitation stress, shear stress, freeze-thaw stress, thermal stress, or any combination therein.

[0153] 25. The formulation of either one of alternatives 23 or 24, wherein the one or more amino acids increases the agitation stress resistance of the formulation as compared to the agitation stress resistance of the same formulation in the absence of the one or more amino acids.

[0154] 26. The formulation of any one of alternatives 23-25, wherein the one or more amino acids increases the freeze-thaw stress resistance of the same formulation as compared to the freeze-thaw stress resistance of the same formulation in the absence of the one or more amino acids.

[0155] 27. The formulation of any one of alternatives 3-26, wherein the detergent increases the thermal stress resistance of the formulation as compared to the thermal stress resistance of the same formulation in the absence of the detergent.

[0156] 28. A formulation, the formulation comprising: a buffer, wherein the buffer is present in an amount ranging between 5 mM and 30 mM; one or more salts, wherein the one or more salts are individually present in an amount ranging between 0.1 mM and 250 mM; arginine and / or proline, wherein the arginine and / or proline are individually present in an amount ranging between 20 and 70 mM; and a detergent, wherein the detergent is present in an amount ranging between 0.0001% and 1%; and wherein the formulation is at a pH ranging between 7.5 and 8.5.

[0157] 29. The formulation of alternative 28, wherein the buffer is selected from the group comprising: acetate, citrate, histidine, phosphate, succinate, Tris, Tris HC1, or any combination thereof.

[0158] 30. The formulation of alternative 28 or 29, wherein the buffer comprisesTris or Tris and Tris HC1 in combination.

[0159] 31. The formulation of any one of alternatives 28-30, wherein the buffer comprises between 5-25 mM Tris or Tris and Tris HC1 in combination.

[0160] 32. The formulation of any one of alternatives 28-30, wherein the buffer comprises 20 mM Tris, 10 mM Tris, 20 20 mM Tris and Tris-HCl in combination, or 10 mM Tris and Tris HC1 in combination.

[0161] 33. The formulation of any one of alternatives 28-32, wherein the one or more salt comprises MgCh, NaCl, or a combination thereof.

[0162] 34. The formulation of any one of alternatives 28-33, wherein the one or more salts comprises between 0.1 and 5 mM MgCh.

[0163] 35. The formulation of any one of alternatives 28-33, wherein the one or more salt comprises between 50 and 250 mM NaCl.

[0164] 36. The formulation of any one of alternatives 28-33, wherein the one or more salt comprises between 0.1 and 5 mM MgCh and between 50 and 250 mM NaCl.

[0165] 37. The formulation of any one of alternatives 28-40, wherein the formulation comprises between 25 and 75 mM arginine.

[0166] 38. The formulation of any one of alternatives 28-41, wherein the formulation comprises between 25 and 75 mM proline.

[0167] 39. The formulation of any one of alternatives 28-40, wherein the formulation comprises between 25 and 75 mM arginine and between 25 and 75 mM proline.

[0168] 40. The formulation of any one of alternatives 28-44, wherein the detergent comprises Pl 88, PS80, or a combination thereof.

[0169] 41. The formulation of one of alternatives 28-45, wherein the detergent comprises about 0.005% P188.

[0170] 42. The formulation of one of alternatives 28-46, wherein the formulation is at a pH ranging between about 7.8 and 8.2

[0171] 43. The formulation of one of alternatives 28-47, wherein the formulation is at a pH of about 8.

[0172] 44. The formulation of alternative 28, wherein the buffer comprises between5 to 25mM Tris or Tris HC1, the one or more salts comprises between 0.5 to 1.5 mM MgCh, the one or more amino acids comprises between 25-75 mM arginine and between 25-75 mM proline, and the detergent comprises between 0.001-0.2% Pl 88, wherein the formulation is at a pH of 7.7-8.3.

[0173] 45. The formulation of alternative 28, wherein the buffer comprises 5 to 25 mM Tris or Tris HCL, the one or more salts comprises between 0.5 and 1.5 mM MgCh and between 50-250 mM NaCl, the one or more amino acids comprises between 25-75 mMarginine and between 25-75 mM proline, and the detergent comprises between 0.001 -0.2% P188, wherein the formulation is at a pH of between 7.7-8.3.

[0174] 46. The formulation of any one of alternatives 28-45, wherein the one or more amino acids increases a stress resistance of the formulation as compared to the stress resistance of the same formulation in the absence of the one or more amino acids.

[0175] 47. The formulation of alternative 46, wherein the stress resistance is selected from agitation stress, shear stress, freeze-thaw stress, thermal stress, or any combination therein.

[0176] 48. The formulation of either one of alternatives 46 or 47, wherein the one or more amino acids increases the agitation stress resistance of the formulation as compared to the agitation stress resistance of the same formulation in the absence of the one or more amino acids.

[0177] 49. The formulation of either one of alternatives 46 or 47, wherein the one or more amino acids increases the freeze-thaw stress resistance of the formulation as compared to the freeze-thaw stress resistance of the same formulation in the absence of the one or more amino acids.

[0178] 50. The formulation of either one of alternatives 46 or 47, wherein the detergent increases the thermal stress resistance of the formulation as compared to the thermal stress resistance of the same formulation in the absence of the detergent.

[0179] 51. The formulation of any one of the preceding alternatives, further comprising a stabilizer.

[0180] 52. The formula of alternative 51, wherein the stabilizer is selected from the group comprising HAS, cyclodextrin, sucrose, sorbitol, or any combination thereof.

[0181] 53. The formulation of alternative 51 or 52, wherein the stabilizer is present in an amount ranging between 0.1% and 20%.

[0182] 54. The formulation of any one of alternatives 51-53, wherein the stabilizer is present in an amount ranging between 0.1 mM and 200 mM.

[0183] 55. The formulation of any one of the preceding alternatives, further comprising about 180 mM sorbitol.

[0184] 56. The formulation of any one of the preceding alternatives, wherein the formulation remains stable at temperatures ranging between -80°C and 20 °C.

[0185] 57. The formulation of any one of the preceding alternatives, wherein the formulation remains stable at -20°C.

[0186] 58. The formulation of any one of the preceding alternatives, wherein the formulation remains stable at temperatures ranging between 2-8°C.

[0187] 59. The formulation of any one of the preceding alternatives, wherein the formulation remains stable for at least 5 freeze-thaw cycles.

[0188] 60. The formulation of any one of the preceding alternatives, wherein the formulation further comprises an AAV vector.

[0189] 61. The formulation of alternative 60, wherein the AAV is an AAV serotype 1 (AAV-1), AAV serotype 2 (AAV-2), AAV serotype 3 (AAV-3), AAV serotype 4 (AAV-4), AAV serotype 5 (AAV-5), AAV serotype 6 (AAV-6), AAV serotype 7 (AAV-7), AAV serotype 8 (AAV-8), AAV serotype 9 (AAV-9), serotype rhlO AAV, serotype rh74 AAV, or a pseudotyped rAAV (e.g., AAV2 / 9, referring an AAV vector with the genome of AAV2 (e.g., the ITRs of AAV2) and the capsid of AAV9).

[0190] 62. The formulation of alternatives 60 or 61, wherein the AAV vector comprises an AAV serotype 9, or is derived from an AAV serotype 9 vector.

[0191] 63. The formulation of any one of alternatives 60-62, wherein the AAV vector comprises an AAV serotype rh74, or is derived from an AAV serotype rh74 vector.

[0192] 64. The formulation of any one of the preceding alternatives, further comprising an expression cassette.

[0193] 65. The formulation of alternative 64, wherein the expression cassette is a therapeutic expression cassette.

[0194] 66. The formulation of alternative 65, wherein the therapeutic expression cassette encodes a therapeutic protein.

[0195] 67. The formulation of alternative 66, wherein the therapeutic protein is selected from the group consisting of frataxin, GSD1A, PRKAG2, BAG3, Dysferlin, MicroDys, MYBPC-3, Danon / Lamp2, TMEM43, PKP2, LMNA, TNNT2, RBM20, MYH7, RYR2, CAMKII, PRKAG2, CASQ2, DM1, GRN, GBA, MSH3, UNC13A, Stathmin-2, SMEI, and / or any combination(s) thereof.

[0196] 68. A method of treatment, the method comprising identifying a subject as likely to benefit from treatment, and administering the formulation of any one of the preceding alternatives.

[0197] 69. The method of alternatives 68, wherein formulation comprises a nucleic acid encoding a therapeutic protein selected from the group consisting of frataxin, GSD1A, PRKAG2, BAG3, Dysferlin, MicroDy s, MYBPC-3, Danon / Lamp2, TMEM43, PKP2, LMNA, TNNT2, RBM20, MYH7, RYR2, CAMKII, PRKAG2, CASQ2, DM1, GRN, GBA, MSH3, UNCI 3 A, Stathmin-2, SMEI, and / or any combination(s) thereof.

[0198] 70. Use of the pharmaceutical formulation of any one of alternatives 1-67, in the manufacture of a medicament for the treatment of disease.

[0199] 71. The use of alternative 70, wherein the formulation comprises a nucleic acid encoding a therapeutic protein selected from the group consisting of frataxin, GSD1A, PRKAG2, BAG3, Dysferlin, MicroDy s, MYBPC-3, Danon / Lamp2, TMEM43, PKP2, LMNA, TNNT2, RBM20, MYH7, RYR2, CAMKII, PRKAG2, CASQ2, DM1, GRN, GBA, MSH3, UNC13A, Stathmin-2, SMEI, and / or any combination(s) thereof.

[0200] 72. A method of manufacturing a formulation, the method comprising combining: a buffer, wherein the buffer is present in an amount ranging between 5 mM and 25 mM; one or more salts, wherein the one or more salts are individually present in an amount ranging between 0.1 mM and 250 mM; one or more amino acids, wherein the one or more amino acids are individually present in an amount ranging between 1 and 200 mM; and a detergent, wherein the detergent is present in an amount ranging between 0.0001% and 1%; and adjusting the formulation to a pH ranging between 7.5 and 8.5.Example 1: Description of a non-limiting dosage form

[0201] In a non-limiting example, the drug product (DP) is a single-dose, sterile, preservative-free suspension for intravenous infusion. DP is supplied in a 10 mL vial with a 9.0 mL deliverable volume. The DP contains a target concentration of 2.0E13vg / mL viral vector with an upper threshold of 2.7E13vg / ml in a formulation of 20 mM Tris buffer containing 1 mM magnesium chloride, 50 mM L proline, 50 mM L arginine, and 0.005% poloxamer 188 at pH 8.0 with a final osmolality of approximately 170 mOsm / kg. DP is stored frozen at < -60°C.

[0202] Drug substance (DS) is a recombinant adeno associated virus (rAAV) gene therapy which can be a modified AAV capsid serotype. Each batch of DP is manufactured to a target vector genome concentration to achieve the desired concentration of DP. If required, the vector may be diluted with final formulation buffer to achieve the desired target vector genome titer. The formulation of the vector comprises a 20 mM Tris buffer containing 1 mM magnesium chloride, 50 mM L proline, 50 mM L arginine, and 0.005% pol oxamer 188 at pH 8.0.

[0203] Excipients in the DP formulation are included for buffering capacity and tonicity, and poloxamer 188 is added to minimize adsorptive losses. The excipients present in the drug product are commonly used in the pharmaceutical industry and are considered safe for parenteral formulations.

[0204] The composition of DP, including quality standards, quantities, and function of each component, is provided in Table 2.Table 2Abbreviations: BP = British Pharmacopoeia, NF = National Formulary, Ph. Eur. = European Pharmacopoeia, q.s. = quantum sufficit, USP = United States Pharmacopeia, vg = vector genomes Note: The vial has an overfill of 0.3 mL to ensure the stated deliverable volume.

[0205] Drug product (DP) may be administered to patients via intravenous (IV) infusion using an IV bag and infusion system with tubing and in-line filter.

[0206] The DP vector genome titer release specification, clinical trial dose, clinical trial patient weight range, and clinical infusion rate can be used to calculate the minimum and maximum dose volumes as well as maximum infusion time. These figures can then be used to select DP concentration, DP fill volume in IV bag, and DP exposure to IV bag time values.

[0207] The minimum dose volume of DP may be based on a IE14vg / kg dose of DP across a patient weight range of 10-30 kg and a DP concentration range of 1.0E13- 2.4E13vg / mL. The lowest patient weight of 10 kg and highest DP concentration of 2.4E13vg / mL would provide a DP volume of 49 m when accounting for dilution from 6 parts DP to 1 part 3% saline clinical formulation procedure.

[0208] For DP formulations in which osmolality is adjusted prior to administration at a ratio of 6: 1 DP to tonicifier (e.g., 3% saline), 6 vials of viral vector DP are allowed to fully thaw, pooled, and then formulated for administration by adding 1 part 3% saline to 6 parts DP, and transferred into and IV bag.

[0209] DP is administered via intravenous infusion. The administered volume of DP is pooled from a sufficient number of vials from a single DP lot. Prior to infusion, DP is clinically formulated to achieve physiological osmolarity levels by dilution in a hypertonic saline solution sourced by the pharmacy. Post-infusion, a normal saline flush is passed through the IV bag and infusion set tubing including an inline filter.Example 2: AAV Stability

[0210] In this non-limiting example, the effects of the formulation buffers (Table 3) on AAV stability in response to forced degradation conditions were evaluated. The AAV reporter, AAV-SLBIOI-Luc, which is the luciferase gene encapsidated within the AAV- SLB101 capsid, was used for these studies.Table 3. Tested Formulation Buffers and Virus Titers

[0211] AAV-SLB-101-Luc was concentrated to approximately 6E13vg / mL and diafiltered into a base buffer (20 mM Tris pH 8, 1 mM magnesium chloride, and 0.005% P188). This material was split into three equal volumes, one for each formulation in Table 3. Each volume was spiked with a concentrated combination of the remaining excipients and diluted with final formulation buffer to approximately 3E13vg / mL. A third of this volume was reserved as a working stock for studies of the 3E13vg / mL titer. The remaining two thirds were diluted with final formulation buffer as the working stock to test the 2E13vg / mL titer.Freeze Thaw Analysis

[0212] Freeze-thaw cycling between -80°C and ambient was performed on working stock aliquots of formulations Fl, F2 and F3 for 5 cycles (5x FT). One cycle consisted of at least 60 minutes at -80°C followed by incubation at room temperature until aliquot was completely thawed. After completion of the final thaw, working stock aliquots were stored at -80°C while awaiting sub-aliquoting for testing.

[0213] FIG. 1A shows, by size exclusion chromatography (SEC) analysis, that AAVs in formulation 1 (Fl), which includes arginine and proline but no NaCl, and formulation 2 (F2) which includes arginine and proline and 61 mM NaCl, are less susceptible to freeze / thaw stress compared AAVs in formulation 3 (F3) which does not contain arginine and proline and has 135 mM NaCl.

[0214] AAVs formulated in Fl and F2 are also more stable compared to F3 formulated viruses as determined by DNA labeling analysis (FIG. IB).Oxidation Analysis

[0215] Working stock aliquots of AAVs formulated in Fl, F2, andF3 were exposed to oxidative stress by combining sample material with a final concentration of 0.3% (v / v) H2O2 at 25°C for 24 hours + / - 1 hour and subsequently quenched with a molar equivalent of Methionine. Upon completion, working stock aliquots were stored at -80°C while awaiting sub-aliquoting for testing. Oxidation was performed concurrently with the oxidation control samples (Ox Ctrl) in which purified water and methionine was substituted for H2O2 and quenching methionine.

[0216] In the SEC assay, the oxidation control was consistent with the results at TO (FIG. 2A). All three formulations exhibited increased HMW content (FIG. 2A). All three formulations also had increased amounts of labelled DNA in the DNA labeling assay (FIG. 2B). However, these effects were most notable in F3 (FIGs. 2A-2B). AAVs formulated in Fl and F2, which include arginine and proline, exhibited increased stability in the oxidation assay compared to F3 which does not include arginine and proline.Thermal Stress (45°C) Analysis

[0217] Working stock aliquots of AAVs formulated in Fl, F2, and F3 were exposed to thermal stress through incubation at 45°C for 0 days, 3 days, and 7 days. Results are presented in FIG. 3A-3C. In each case, the formulation with the lowest tonicity (Fl) showed the most sensitivity to thermal stress. F2 and F3, with higher osmolalities, proved more resilient.

[0218] Although the foregoing has been described in some detail by way of illustrations and examples for purposes of clarity and understanding, it will be understood by those of skill in the art that numerous and various modifications can be made without departing from the spirit of the present disclosure. Therefore, it should be clearly understood that the forms disclosed herein are illustrative only and are not intended to limit the scope of the present disclosure, but rather to also cover all modification and alternatives coming with the true scope and spirit of the disclosure.

Claims

WHAT IS CLAIMED IS:

1. A formulation, the formulation comprising: (a) 5 to 25 mM Tris, (b) 0.5-1.5 mM MgC12, (c) 25-75 mM arginine, (d) 25-75 mM proline, and (e) 0.001-0.2% P188, wherein the formulation is at a pH of 7.7-8.3.

2. A formulation, the formulation comprising: (a) 5 to 25 mM Tris, (b) 0.5-1.5 mM MgC12, (c) 50-250 mM NaCl, (d) 25-75 mM arginine, (e) 25-75 mM proline, and (f) 0.001- 0.2% P188, wherein the formulation is at a pH of 7.7-8.3.

3. A formulation, the formulation comprising: a) a buffer, wherein the buffer is present in an amount ranging between 5 mM and 30 mM; b) one or more salts, wherein the one or more salts are individually present in an amount ranging between 0.1 mM and 200 mM; c) one or more amino acids, wherein the one or more amino acids are individually present in an amount ranging between 1 and 200 mM; and d) a detergent, wherein the detergent is present in an amount ranging between 0.0001% and 1%; and wherein the formulation is at a pH ranging between 7.5 and 8.5.

4. The formulation of claim 3, wherein the buffer is selected from the group comprising: acetate, citrate, histidine, phosphate, succinate, Tris, and Tris-HCl, or any combination thereof.

5. The formulation of claim 3, wherein the buffer comprises Tris or Tris HCL.

6. The formulation of any one of claims 3-5, wherein the buffer comprises 5 to 25 mM Tris or Tris and Tris HCL in combination.

7. The formulation of claim 3, wherein the buffer comprises: 20 mM Tris, 10 mM Tris, 20 mM Tris and Tris-HCl in combination, or 10 mM Tris and Tris HCL in combination.

8. The formulation of claim 3, wherein the one or more salt comprises MgCh, NaCl, or a combination thereof.

9. The formulation of claim 3, wherein the one or more salts comprises between 0.1 and 5 mM MgCh.

10. The formulation of claim 3, wherein the one or more salt comprises between 50 and 250 mM NaCl.

11. The formulation of claim 8, wherein the one or more salt comprises 1 mM MgCh and 75 mM NaCl.

12. The formulation of claim 8, wherein the one or more salt comprises 1 mM MgCh and 200 mM NaCl.

13. The formulation of claim 3, wherein the one or more amino acids are selected from the group comprising: alanine, arginine, glutamic acid, glycine, methionine, proline, serine, or any combination thereof.

14. The formulation of claim 3, wherein the one or more amino acids comprises between 25 and 75 mM arginine.

15. The formulation of claim 3, wherein the one or more ammo acids comprises between 25 and 75 mM proline.

16. The formulation of claim 3, wherein the one or more amino acids comprises between 25 and 75 mM arginine and between 25 and 75 mM proline.

17. The formulation of claim 3, wherein the detergent comprises P188, PS80, or a combination thereof.

18. The formulation of claim 3, wherein the detergent comprises between 0.001 and 0.01% P188.

19. The formulation of claim 3, wherein the formulation is at a pH ranging between 7.7 and 8.3.

20. The formulation of claim 3, wherein the formulation is at a pH of 8.

21. The formulation of claim 3, wherein the buffer comprises 5 to 25 mM Tris or Tris HC1, the one or more salts comprises 0.5-1.5 mM MgCh, the one or more amino acids comprises 25-75 mM arginine and 25-75 mM proline, and the detergent comprises 0.001-0.2% P188, wherein the formulation is at a pH of 7.7-8.3.

22. The formulation of claim 3, wherein the buffer comprises 5 to 30 mM Tris or Tris HC1, the one or more salts comprises 0.5-1.5 mM MgCh and 50-250 mM NaCl, the one or more amino acids comprises 30-60 mM arginine and 30-60 mM proline, and the detergent comprises 0.001-0.2% P188, wherein the formulation is at a pH of 7.7-8.3.

23. The formulation of claim 3, wherein the one or more amino acids increases a stress resistance of the formulation as compared to the stress resistance of the same formulation in the absence of the one or more amino acids.

24. The formulation of claim 23, wherein the stress resistance is selected from agitation stress, shear stress, freeze-thaw stress, thermal stress, or any combination therein.

25. The formulation of claim 23, wherein the one or more amino acids increases the agitation stress resistance of the formulation as compared to the agitation stress resistance of the same formulation in the absence of the one or more amino acids.

26. The formulation of claim 23, wherein the one or more amino acids increases the freeze-thaw stress resistance of the same formulation as compared to the freeze-thaw stress resistance of the same formulation in the absence of the one or more amino acids.

27. The formulation of claim 3, wherein the detergent increases the thermal stress resistance of the formulation as compared to the thermal stress resistance of the same formulation in the absence of the detergent.

28. A formulation, the formulation comprising: a) a buffer, wherein the buffer is present in an amount ranging between 5 mM and 30 mM; b) one or more salts, wherein the one or more salts are individually present in an amount ranging between 0.1 mM and 250 mM; c) arginine and / or proline, wherein the arginine and / or proline are individually present in an amount ranging between 20 and 70 mM; and d) a detergent, wherein the detergent is present in an amount ranging between 0.0001% and 1%; and wherein the formulation is at a pH ranging between 7.5 and 8.5.

29. The formulation of claim 28, wherein the buffer is selected from the group comprising: acetate, citrate, histidine, phosphate, succinate, Tris, Tris HCl, or any combination thereof.

30. The formulation of claim 28, wherein the buffer comprises Tris or Tris and Tris HC1 in combination.

31. The formulation of claim 28, wherein the buffer comprises between 5-25 mM Tris or Tris and Tris HC1 in combination.

32. The formulation of claim 28, wherein the buffer comprises 20 mM Tris, 10 mM Tris, 2020 mM Tris and Tris-HCl in combination, or 10 mM Tris and Tris HC1 in combination.

33. The formulation of claim 28, wherein the one or more salt comprises MgCh, NaCl, or a combination thereof.

34. The formulation of claim 28, wherein the one or more salts comprises between 0.1 and 5 mM MgCh.

35. The formulation of claim 28, wherein the one or more salt comprises between 50 and 250 mM NaCl.

36. The formulation of claim 28, wherein the one or more salt comprises between 0.1 and 5 mM MgCh and between 50 and 250 mM NaCl.

37. The formulation of claim 28, wherein the formulation comprises between 25 and 75 mM arginine.

38. The formulation of claim 28, wherein the formulation comprises between 25 and 75 mM proline.

39. The formulation of claim 28, wherein the formulation comprises between 25 and 75 mM arginine and between 25 and 75 mM proline.

40. The formulation of claim 28, wherein the detergent comprises Pl 88, PS80, or a combination thereof.

41. The formulation of claim 28, wherein the detergent comprises about 0.005% P188.

42. The formulation of claim 28, wherein the formulation is at a pH ranging between about 7.8 and 8.

243. The formulation of claim 28, wherein the formulation is at a pH of about 8.

44. The formulation of claim 28, wherein the buffer comprises between 5 to 25mM Tris or Tris HC1, the one or more salts comprises between 0.5 to 1.5 mM MgCh, the one or more amino acids comprises between 25-75 mM arginine and between 25-75 mM proline, and the detergent comprises between 0.001-0.2% P188, wherein the formulation is at a pH of 7.7- 8.3.

45. The formulation of claim 28, wherein the buffer comprises 5 to 25 mM Tris or Tris HCL, the one or more salts comprises between 0.5 and 1.5 mM MgCh and between 50-250 mM NaCl, the one or more amino acids comprises between 25-75 mM arginine and between25-75 mM proline, and the detergent comprises between 0.001-0.2% Pl 88, wherein the formulation is at a pH of between 7.7-8.3.

46. The formulation of claim 28, wherein the one or more amino acids increases a stress resistance of the formulation as compared to the stress resistance of the same formulation in the absence of the one or more amino acids.

47. The formulation of claim 46, wherein the stress resistance is selected from agitation stress, shear stress, freeze-thaw stress, thermal stress, or any combination therein.

48. The formulation of claim 46, wherein the one or more amino acids increases the agitation stress resistance of the formulation as compared to the agitation stress resistance of the same formulation in the absence of the one or more amino acids.

49. The formulation of claim 46, wherein the one or more amino acids increases the freeze-thaw stress resistance of the formulation as compared to the freeze-thaw stress resistance of the same formulation in the absence of the one or more amino acids.

50. The formulation of claim 46, wherein the detergent increases the thermal stress resistance of the formulation as compared to the thermal stress resistance of the same formulation in the absence of the detergent.

51. The formulation of claim 3, further comprising a stabilizer.

52. The formula of claim 51, wherein the stabilizer is selected from the group comprising HAS, cyclodextrin, sucrose, sorbitol, or any combination thereof.

53. The formulation of claim 51, wherein the stabilizer is present in an amount ranging between 0.1% and 20%.

54. The formulation of claim 51, wherein the stabilizer is present in an amount ranging between 0.1 mM and 200 mM.

55. The formulation of claim 3, further comprising about 180 mM sorbitol.

56. The formulation of claim 3, wherein the formulation remains stable at temperatures ranging between -80°C and 20 °C.

57. The formulation of claim 3, wherein the formulation remains stable at -20°C.

58. The formulation of claim 3, wherein the formulation remains stable at temperatures ranging between 2-8°C.

59. The formulation of claim 3, wherein the formulation remains stable for at least 5 freeze-thaw cycles.

60. The formulation of claim 3, wherein the formulation further comprises an AAV vector.

61. The formulation of claim 60, wherein the AAV is an AAV serotype 1 (AAV-1), AAV serotype 2 (AAV-2), AAV serotype 3 (AAV-3), AAV serotype 4 (AAV-4), AAV serotype 5 (AAV-5), AAV serotype 6 (AAV-6), AAV serotype 7 (AAV-7), AAV serotype 8 (AAV-8), AAV serotype 9 (AAV-9), serotype rhlO AAV, serotype rh74 AAV, or a pseudotyped rAAV (e.g., AAV2 / 9, referring an AAV vector with the genome of AAV2 (e.g., the ITRs of AAV2) and the capsid of AAV9).

62. The formulation of claim 60, wherein the AAV vector comprises an AAV serotype 9, or is derived from an AAV serotype 9 vector.

63. The formulation of claim 60, wherein the AAV vector comprises an AAV serotype rh74, or is derived from an AAV serotype rh74 vector.

64. The formulation of claim 3, further comprising an expression cassette.

65. The formulation of claim 64, wherein the expression cassette is a therapeutic expression cassette.

66. The formulation of claim 65, wherein the therapeutic expression cassette encodes a therapeutic protein.

67. The formulation of claim 66, wherein the therapeutic protein is selected from the group consisting of frataxin, GSD1A, PRKAG2, BAG3, Dysferlin, MicroDys, MYBPC-3, Danon / Lamp2, TMEM43, PKP2, LMNA, TNNT2, RBM20, MYH7, RYR2, CAMKII, PRKAG2, CASQ2, DM1, GRN, GBA, MSH3, UNC13A, Stathmin-2, SMEI, and / or any combination(s) thereof.

68. A method of treatment, the method comprising identifying a subject as likely to benefit from treatment, and administering the formulation of any one of the preceding claims.

69. The method of claim 68, wherein formulation comprises a nucleic acid encoding a therapeutic protein selected from the group consisting of frataxin, GSD1A, PRKAG2, BAG3, Dysferlin, MicroDys, MYBPC-3, Danon / Lamp2, TMEM43, PKP2, LMNA, TNNT2, RBM20, MYH7, RYR2, CAMKII, PRKAG2, CASQ2, DM1, GRN, GBA, MSH3, UNCI 3 A, Stathmin- 2, SMEI, and / or any combination(s) thereof.

70. Use of the pharmaceutical formulation of any one of claims 1-67, in the manufacture of a medicament for the treatment of disease.71 . The use of claim 70, wherein the formulation comprises a nucleic acid encoding a therapeutic protein selected from the group consisting of frataxin, GSD1A, PRKAG2, BAG3, Dysferlin, MicroDys, MYBPC-3, Danon / Lamp2, TMEM43, PKP2, LMNA, TNNT2, RBM20, MYH7, RYR2, CAMKII, PRKAG2, CASQ2, DM1, GRN, GBA, MSH3, UNCI 3 A, Stathmin- 2, SMEI, and / or any combination(s) thereof.

72. A method of manufacturing a formulation, the method comprising combining: a) a buffer, wherein the buffer is present in an amount ranging between 5 mM and 25 mM; b) one or more salts, wherein the one or more salts are individually present in an amount ranging between 0.1 mM and 250 mM; c) one or more amino acids, wherein the one or more amino acids are individually present in an amount ranging between 1 and 200 mM; and d) a detergent, wherein the detergent is present in an amount ranging between 0.0001% and 1%; and adjusting the formulation to a pH ranging between 7.5 and 8.5.

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