SUBCUTANEOUS DELIVERY OF MESSENGER RNA
The present invention provides, among other things, methods of formulating nucleic acid-containing nanoparticles with an enzyme to afford efficient delivery of payload to a cell or tissue of interest via subcutaneous administration. In some embodiments, the present invention provides a process in which mRNA-loaded lipid nanoparticles are co-mixed with various amounts of hyaluronidase and administered via subcutaneous administration. The resulting payload can be efficiently delivered to the liver and other organs or tissues of a treated subject.
1 . A method of treating ornithine transcarbamylase (OTC deficiency) comprising
administering via subcutaneous injection to a subject in need of treatment
a) an mRNA encoding an ornithine transcarbamylase (OTC) protein wherein the mRNA is encapsulated within a lipid based nanoparticle, and
b) a hyaluronidase enzyme at a concentration of at least 200U, wherein the subcutaneous injection results in expression of the OTC protein in the liver of the subject.
2 .- 13 . (canceled)
14 . The method of claim 1 , wherein the lipid based nanoparticle is a liposome comprising a PEGylated lipid
15 . (canceled)
16 . The method of claim 14 , wherein the PEGylated lipid constitutes at least 5% of the total lipids in the liposome.
17 . (canceled)
18 . The method of claim 1 , wherein the subcutaneous injection results in a reduced urinary orotic acid level in the subject as compared to a control orotic acid level, wherein the control orotic acid level is either a baseline urinary orotic acid level in the subject prior to the treatment, or a reference level indicative of the average urinary orotic acid level in OTC patients without treatment.
19 .- 43 . (canceled)
44 . A method of messenger RNA (mRNA) delivery for in vivo protein expression, comprising,
administering via subcutaneous injection to a subject
a) an mRNA encoding a protein wherein the mRNA is encapsulated within a nanoparticle, and
b) a hyaluronidase enzyme at a concentration of at least 200 U, wherein the hyaluronidase enzyme is co-formulated with the mRNA.
45 .- 47 . (canceled)
48 . The method of claim 44 , wherein the protein is detectable after at least 24 hours, 2 days, 3 days, 4 days, 5 days, 6 days, 1 week, 2 weeks, 3 weeks, 4 weeks, or 1 month post-injection.
49 .- 54 . (canceled)
55 . The method of claim 44 , wherein the nanoparticle is a lipid-based or polymer-based nanoparticle.
56 . The method of claim 55 , wherein the lipid-based nanoparticle is a liposome comprising a PEGylated lipid.
57 . (canceled)
58 . The method of claim 56 , wherein the PEGylated lipid constitutes at least 1%, at least 2%, at least 3%, at least 4%, at least 5%, at least 6%, at least 7%, at least 8%, at least 9%, or at least 10% of the total lipids in the liposome
59 . The method of claim 58 , wherein the PEGylated lipid constitutes 5% or less of the total lipids in the liposome.
60 .- 64 . (canceled)
65 . A composition for delivery of mRNA for in vivo protein expression, comprising
a) an mRNA encoding a protein wherein the mRNA is encapsulated within a nanoparticle, and
b) a hyaluronidase enzyme at a concentration of at least 200U, wherein the hyaluronidase enzyme is co-formulated in the mRNA.
66 .- 69 . (canceled)
70 . The composition of claim 65 , wherein the nanoparticle is a lipid-based or polymer-based nanoparticle.
71 . The composition of claim 70 , wherein the lipid-based nanoparticle is a liposome comprising a PEGylated lipid.
72 . (canceled)
73 . The composition of claim 71 , wherein the PEGylated lipid constitutes at least 1%, at least 2%, at least 3%, at least 4%, at least 5%, at least 6%, at least 7%, at least 8%, at least 9%, or at least 10% of the total lipids in the liposome.
74 . The composition of claim 65 , wherein the mRNA comprises one or more modified nucleotides.
75 . The composition of claim 65 , wherein the mRNA is unmodified.
76 . The composition of claim 65 , wherein the composition is in a liquid form.
77 . The composition of claim 65 , wherein the composition is lyophilized powder.
78 .- 79 . (canceled)
80 . A method of treating a disease, disorder or condition comprising delivering messenger RNA (mRNA) to a subject in need of treatment according to a method of claim 44 , wherein the mRNA encodes a protein deficient in the subject.
81 . (canceled)
82 . The method of claim 80 , wherein the disease, disorder or condition is a metabolic disorder.
83 . (canceled)