Peptide-mediated intravesical delivery of therapeutic and diagnostic agents

1. A pharmaceutical composition comprising protamine and an antisense oligonucleotide directed against nerve growth factor, where said composition is for intravesical administration to a subject.

2. The pharmaceutical composition of claim 1 , where protamine comprises one or more of the following sequences:

MARYRCCRSQ SRSRYYRQRQ RSRRRRRRSC QTRRRAMRCC RPRYRPRCRR H (SEQ ID NO:1);

MVRYRVRSLS ERSHEVYRQQ LHGQEQGHHG QEEQGLSPEH VEVYERTHGQ SHYRRRHCSR RRLHRIHRRQ HRSCRRRKRR SCRHRRRHRR GCRTRKRTCR RH (SEQ ID NO:2);

(SEQ ID NO: 1)

MARYRCCRSQ SRSRYYRQRQ RSRRRRRRSC QTRRRAMRCC

RPRYRPRCRR H;

(SEQ ID NO: 2)

MVRYRVRSLS ERSHEVYRQQ LHGQEQGHHG QEEQGLSPEH

VEVYERTHGQ SHYRRRHCSR RRLHRIHRRQ HRSCRRRKRR 

SCRHRRRHRR GCRTRKRTCR RH;

(SEQ ID NO: 3)

MRRQASLPAR RRRRVRRTRV VRRRRRVGRR RH;

(SEQ ID NO: 4)

PRRRRRSSSR PIRRRRRPRAS RRRRGGRRR R;

(SEQ ID NO: 5)

PRRRRSSRRP VRRRRRPRVSR RRRRGGRRRR;

(SEQ ID NO: 6)

PRRRRSSSRP VRRRRRPRVSR RRRRGGRRRR;

(SEQ ID NO: 7)

PRRRRASRRI RRRRRPRVSR RRRRGGRRRR;

and/or

a peptide comprising the sequence RRRX 1 X 2 RRR where X 1 or X 2 or both is/are G or V or A (SEQ ID NO:8).

3. The pharmaceutical composition of claim 1 which is prepared by mixing together protamine and the antisense oligonucleotide directed against nerve growth factor shortly before administration to a subject.

4. A kit for preparing for intravesical administration to a subject, comprising protamine and an antisense oligonucleotide directed against nerve growth factor.

5. The pharmaceutical composition of claim 1 , where the mass ratio of protamine to the antisense oligonucleotide directed against nerve growth factor is between about 5:1 to about 15:1 respectively.

6. The pharmaceutical composition of claim 1 , where the mass ratio of protamine to the antisense oligonucleotide directed against nerve growth factor is about 10:1 respectively.

7. The pharmaceutical composition of claim 1 , where the mass ratio of protamine to the antisense oligonucleotide directed against nerve growth factor is about 5:1 respectively.

8. The pharmaceutical composition of claim 1 , comprising the antisense oligonucleotide directed against nerve growth factor in a concentration range between about 2 to about 20 μM.

9. The pharmaceutical composition of claim 1 , comprising protamine in a concentration range between about 20 to about 200 μM.

10. The pharmaceutical composition of claim 1 , comprising the antisense oligonucleotide directed against nerve growth factor in a concentration range between about 2 to about 20 μM and protamine in a concentration range between about 20 to about 200 μM.

11. The pharmaceutical composition of claim 1 , where the antisense oligonucleotide is directed against nerve growth factor encoded by a mRNA having a corresponding cDNA sequence as follows:

[SEQ ID NO: 9]

   1 tctgctttta ataagcttcc caatcagctc tcgagtgcaa agcgctctcc ctccctcgcc

  61 cagccttcgt cctcctggcc cgctcctctc atccctccca ttctccattt cccttccgtt

 121 ccctccctgt cagggcgtaa ttgagtcaaa ggcaggatca ggttccccgc cttccagtcc

 181 aaaaatcccg ccaagagagc cccagagcag aggaaaatcc aaagtggaga gaggggaaga

 241 aagagaccag tgagtcatcc gtccagaagg cggggagagc agcagcggcc caagcaggag

 301 ctgcagcgag ccgggtacct ggactcagcg gtagcaacct cgccccctgc aacaaaggca

 361 gactgagcgc cagagaggac gtttccaact caaaaatgca ggctcaacag taccagcagc

 421 agcgtcgaaa atttgcagct gccttcttgg cattcatttt catactggca gctgtggata

 481 ctgctgaagc agggaagaaa gagaaaccag aaaaaaaagt gaagaagtct gactgtggag

 541 aatggcagtg gagtgtgtgt gtgcccacca gtggagactg tgggctgggc acacgggagg

 601 gcactcggac tggagctgag tgcaagcaaa ccatgaagac ccagagatgt aagatcccct

 661 gcaactggaa gaagcaattt ggcgcggagt gcaaatacca gttccaggcc tggggagaat

 721 gtgacctgaa cacagccctg aagaccagaa ctggaagtct gaagcgagcc ctgcacaatg

 781 ccgaatgcca gaagactgtc accatctcca agccctgtgg caaactgacc aagcccaaac

 841 ctcaagcaga atctaagaag aagaaaaagg aaggcaagaa acaggagaag atgctggatt

 901 aaaagatgtc acctgtggaa cataaaaagg acatcagcaa acaggatcag ttaactattg

 961 catttatatg taccgtaggc tttgtattca aaaattatct atagctaagt acacaataag

1021 caaaaacaa.

12. The pharmaceutical composition of claim 1 , where the antisense oligonucleotide directed against nerve growth factor comprises the following sequence: GCCCGAGACGCCTCCCGA (SEQ ID NO: 10).