Induction of osteogenesis by delivering BMP encoding RNA
The present invention relates to a pharmaceutical composition comprising a polyribonucleotide (RNA) with a sequence which encodes a bone morphogenetic protein (BMP) for use in (i) treating or preventing a bone disease, bone disorder or bone injury; and/or (ii) inducing or enhancing osteogenic differentiation, osteogenesis, ossification, bone regeneration and/or bone morphogenesis in a patient. The present invention also relates to the respective BMP encoding RNAs (BMP RNAs), in particular in its chemically modified form. The present invention also relates to complexes which comprise or are complexed with the BMP RNA, in particular to the respective transfection complexes like lipofection, magnetofection and magnetolipofection complexes. The present invention further relates to a carrier and carrier body to which the RNA or complex has been loaded and to a pharmaceutical composition comprising said carrier or carrier body. The present invention further relates to a matrix or scaffold for sustained mRNA delivery and its application in bone regeneration in vivo, ex vivo and in vitro.
1. A method of treating or preventing a bone disease, bone disorder or bone injury comprising administering to a patient in need thereof a pharmaceutical composition comprising (i) a polyribonucleotide (RNA) with a sequence which encodes a bone morphogenetic protein (BMP); and (ii) a component comprising an oligo (alkylene amine) being a cationic lipidoid having the structure of formula (IV):
wherein the variables a, b, p, m, n and R 1 to R 6 are defined as follows:
a is 1 and b is 2;
p is 1,
m is 1 and n is 1; and
R 1 to R 6 are independently of each other hydrogen, —CH 2 —CH(OH)—R 7 , —CH(R 7 )—CH 2 —OH, —CH 2 —CH 2 —(C═O)—NH—R 7 or —CH 2 —R 7 , wherein R 7 is C3-C18 alkyl or C3-C18 alkenyl having one C—C double bond; provided that one of R 1 , R 2 , R 5 , and R 6 is hydrogen; and wherein one or more of the nitrogen atoms indicated in formula (IV) may be protonated.
2. The method of claim 1 , wherein said BMP is BMP-2 or BMP-7.
3. The method of claim 1 , wherein said RNA forms a complex with said cationic lipidoid.
4. The method of claim 1 , wherein said RNA is delivered in vivo.
5. The method of claim 1 , wherein said RNA is delivered ex vivo into cells which are to be introduced into said patient.
6. The method of claim 1 , wherein said RNA is administered into or in close proximity to the tissue of said patient in which induction of bone growth is desired.
7. The method of claim 5 , wherein said cells are mesenchymal stem cells (MSCs) or osteoprogenitor cells.
8. The method of claim 7 , wherein said MSCs are adipose-derived mesenchymal stem cells (AMSCs) or bone marrow-derived MSCs (BMSCs).
9. The method of claim 1 , wherein the pharmaceutical composition further comprises a matrix or scaffold to which said RNA has been added or into which said RNA has been loaded.
10. The method of claim 9 , wherein said matrix or scaffold comprises collagen and/or fibrin.
11. The method of claim 9 , wherein said matrix or scaffold is vacuum-dried.
12. The method of claim 5 , wherein said cells have been seeded on a matrix or scaffold.
13. The method of claim 9 , wherein said matrix or scaffold is transplanted into the bone of said patient.
14. The method of claim 1 , wherein the RNA is chemically modified, and wherein 25% of the cytidines of said chemically modified RNA are 5-methylcytidines (m5C) and 25% of the uridines of said chemically modified RNA are 2-thiouridines (s2U).
15. The method of claim 1 , wherein said RNA is a single-stranded RNA.
16. The method of claim 1 , wherein said pharmaceutical composition further comprises one or more of cholesterol, DPPC, DOPE and PEG-lipids.
17. The method of claim 1 , wherein said pharmaceutical composition further comprises
(i) DOPE, cholesterol and DMPE-PEG (2k); or
(ii) DPPC, cholesterol and DMG-PEG (2k).
18. The method of claim 17 , wherein the molar ratios of said lipidoid and (i) or (ii) are 8/5.3/4.4/0.9 or 8:5.29:4.41:0.88.
19. A method of treating or preventing a bone disease, bone disorder or bone injury comprising administering to a patient in need thereof a pharmaceutical composition comprising (i) a polyribonucleotide (RNA) with a sequence which encodes a bone morphogenetic protein (BMP); (ii) dipalmitoyl phosphatidylcholine (DPPC) and (iii) a component comprising an oligo (alkylene amine) being a cationic lipidoid having the structure of formula (IV):
wherein the variables a, b, p, m, n and R1 to R6 are defined as follows:
a is 1 and b is 2;
p is 1,
m is 1 and n is 1; and
R 1 to R 6 are independently of each other hydrogen, —CH 2 —CH(OH)—R 7 , —CH(R 7 )—CH 2 —OH, —CH 2 —CH 2 —(C═O)—O—R 7 , —CH 2 —CH 2 —(C═O)—NH—R 7 or —CH 2 —R 7 , wherein R 7 is C3-C18 alkyl or C3-C18 alkenyl having one C—C double bond; provided that one of R 1 , R 2 , R 5 , and R 6 is hydrogen; and wherein one or more of the nitrogen atoms indicated in formula (IV) may be protonated.
20. A method of treating or preventing a bone disease, bone disorder or bone injury comprising administering to a patient in need thereof a pharmaceutical composition comprising (i) a polyribonucleotide (RNA) with a sequence which encodes a bone morphogenetic protein (BMP); and (ii) a component comprising an oligo (alkylene amine) prepared by mixing N,N′-Bis (2-aminoethyl)-1,3-propanediamine with 1,2-Epoxydodecane in a molar ratio of 1:5, and wherein the cationic lipidoid thus obtained has being a cationic lipidoid having the structure of formula (IV):
wherein the variables a, b, p, m, n and R 1 to R 6 are defined as follows:
a is 1 and b is 2;
p is 1,
m is 1 and n is 1; and
R 1 to R 6 are independently of each other selected from hydrogen, CH 2 —CH (OH)—R 7 , and —CH(R 7 )—CH 2 —OH, wherein R 7 is C10 alkyl; provided that five of R 1 to R 6 are selected from CH 2 —CH(OH)—R 7 and —CH(R 7 )—CH 2 —OH, and one of R 1 , R 2 , R 5 , and R 6 is hydrogen; and wherein one or more of the nitrogen atoms indicated in formula (IV) may be protonated.