Claims for Patent: 6,492,106

Summary for Patent: 6,492,106

Title:	Mammalian proteins that bind to FKBP12 in a rapamycin-dependent fashion
Abstract:	A protein complex containing 245 kDa and 35 kDa components, designated RAFT1 and RAFT2 (for Rapamycin And FKBP12 Target) interacts with FKBP12 in a rapamycin-dependent manner. This interaction has the pharmacological characteristics expected from the observed in vivo effects of rapamycin: it occurs at low nanomolar concentrations of rapamycin and is competed by excess FK506. Sequences (330 amino acids total) of tryptic peptides derived from the affinity purified 245 kDa RAFT1 reveals striking homologies to the predicted products of the yeast TOR genes, which were originally identified by mutations that confer rapamycin resistance in yeast. A RAFT1 cDNA was obtained and found to encode a 289 kDa protein (2550 amino acids) that is 43% and 39% identical to TOR2 and TOR1, respectively.
Inventor(s):	Sabatini; David M. (Baltimore, MD), Erdjument-Bromage; Hediye (New York, NY), Lui; Mary (Kew Gardens, NY), Tempst; Paul (New York, NY), Snyder; Solomon H. (Baltimore, MD)
Assignee:	The Johns Hopkins University (Baltimore, MD)
Application Number:	08/305,790
Patent Claims:	1. An isolated, purified cDNA molecule which encodes RAFT1, a protein having the amino acid sequence as shown in SEQ ID NO:2 wherein the acronym RAFT connotes a rapamycin and FKBP12 target. 2. The isolated, purified cDNA molecule of claim 1 which comprises the nucleotide sequence as shown in SEQ ID NO:1, nucleotides 64-7710. 3. An isolated, purified intron-free DNA molecule consisting of at least 20 contiguous nucleotides encoding all or a portion of the amino acid sequence as shown in SEQ ID NO: 2. 4. An isolated, purified intron-free DNA molecule consisting of at least 20 contiguous nucleotides of the sequence as shown in SEQ ID NO: 1. 5. An isolated DNA molecule encoding a rat RAFT protein obtained by a method comprising the steps of: (a) probing a library of rat cDNA sequences with a probe which comprises at least 15 contiguous nucleotides selected from the sequence shown in SEQ ID NO: 1; and (b) isolating a rat cDNA molecule which (i) hybridizes to the probe, (ii) contains a complete open reading frame encoding a polypeptide of about 2550 amino acids, and (iii) encodes a rat RAFT protein, wherein said rat RAFT protein binds to FKBP12 in the presence of 1 to 10 nM rapamycin but not in the absence of 1 to 10 nM rapamycin. 6. An isolated DNA molecule encoding a rat RAFT protein obtained by a method comprising the steps of: (a) amplifying a DNA sequence using (i) at least one primer which comprises at least 10 contiguous nucleotides selected from the sequence shown in SEQ ID NO: 1 and (ii) a template which comprises rat cDNA or mRNA; and (b) isolating an amplified DNA sequence which contains a complete open reading frame encoding a polypeptide of about 2550 amino acids encoding a rat RAFT protein, wherein said rat RAFT protein binds to FKBP12 in the presence of 1 to 10 nM rapamycin but not in the absence of 1 to 10 nM rapamycin. 7. An isolated DNA molecule encoding a rat RAFT protein identified by a process comprising the steps of: (a) annealing a set of mixed oligonucleotides to a rat cDNA library, each member of said set of mixed oligonucleotides encoding a sequence of at least six contiguous amino acids of the amino acid sequence shown in SEQ ID NO:2; and (b) isolating a rat cDNA molecule which (i) anneals to at least one member of the set of mixed oligonucleotides, (ii) contains a complete open reading frame encoding a polypeptide of about 2550 amino acids, and (iii) encodes a rat RAFT protein, wherein said RAFT protein binds to FKBP12 in the presence of 1 to 10 nM rapamycin but not in the absence of 1 to 10 nM rapamycin. 8. An isolated DNA molecule encoding a rat RAFT protein according to claim 7, wherein two sets of mixed oligonucleotides are annealed. 9. An isolated DNA molecule having a nucleotide sequence, or a degenerate sequence thereof, obtained by a method comprising the steps of: (a) probing a library of rat cDNA molecules with a probe which comprises at least 15 contiguous nucleotides selected from the sequence shown in SEQ ID NO: 1; and (b) isolating a rat cDNA molecule which (i) hybridizes to the probe, (ii) contains a complete open reading frame encoding a polypeptide of about 2550 amino acids, and (iii) encodes a rat RAFT protein, wherein said RAFT protein binds to FKBP12 in the presence of 1 to 10 nM rapamycin but not in the absence of 1 to 10 nM rapamycin. 10. A method of isolating a DNA molecule encoding a mammalian RAFT protein comprising the steps of: (a) probing a library of rat cDNA sequences with a probe which comprises at least 15 contiguous nucleotides selected from the sequence shown in SEQ ID NO: 1; and (b) isolating a rat cDNA molecule which (i) hybridizes to the probe, (ii) contains a complete open reading frame encoding a polypeptide of about 2550 amino acids, and (iii) encodes a rat RAFT protein, wherein said rat RAFT protein binds to FKBP12 in the presence of 1 to 10 nM rapamycin but not in the absence of 1 to 10 nM rapamycin. 11. The method of claim 10 wherein the probe comprises at least 20 contiguous nucleotides encoding all or a portion of the amino acid sequence as shown in SEQ ID NO:2. 12. The method of claim 10 wherein the probe comprises at least 20 contiguous nucleotides as shown in SEQ ID NO:1.

Details for Patent 6,492,106

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Merck Sharp & Dohme Corp.	INTRON A	interferon alfa-2b	For Injection	103132	06/04/1986		2019-12-10
Merck Sharp & Dohme Corp.	INTRON A	interferon alfa-2b	For Injection	103132			2019-12-10
Merck Sharp & Dohme Corp.	INTRON A	interferon alfa-2b	Injection	103132			2019-12-10