Polymer reversal on solid surfaces

• US 5,242,974 A
• Filed: 11/22/1991
• Issued: 09/07/1993
• Est. Priority Date: 11/22/1991
• Status: Expired due to Term

• Current Assignee: Affymetrix, Inc. (Thermo Fisher Scientific Incorporated)
• Original Assignee: Affymax Incorporated
• Inventors: Holmes, Christopher P.
• Primary Examiner(s): Nutter, Nathan M.
• Application Number:
  US07/796,727
• Time in Patent Office: 655 Days
• Field of Search: 525/54.11, 530/333, 530/334, 530/335, 530/336, 530/337
• US Class Current: 525/54.11
• CPC Class Codes: B01J 2219/00432 Photolithographic masks
• CPC Class Codes: B01J 2219/00527 Sheets
• CPC Class Codes: B01J 2219/00529 DNA chips
• CPC Class Codes: B01J 2219/00585 Parallel processes
• CPC Class Codes: B01J 2219/0059 Sequential processes
• CPC Class Codes: B01J 2219/00596 Solid-phase processes
• CPC Class Codes: B01J 2219/00605 the compounds being directly bound or immobilised to solid supports
• CPC Class Codes: B01J 2219/00608 DNA chips
• CPC Class Codes: B01J 2219/0061 The surface being organic
• CPC Class Codes: B01J 2219/00612 the surface being inorganic
• CPC Class Codes: B01J 2219/00617 by chemical means
• CPC Class Codes: B01J 2219/00626 Covalent
• CPC Class Codes: B01J 2219/00659 Two-dimensional arrays
• CPC Class Codes: B01J 2219/00711 Light-directed synthesis
• CPC Class Codes: B01J 2219/00722 Nucleotides
• CPC Class Codes: B01J 2219/00725 Peptides
• CPC Class Codes: B82Y 30/00 Nanotechnology for materials or surface science, e.g. nanocomposites
• CPC Class Codes: C07B 2200/11 Compounds covalently bound to a solid support
• CPC Class Codes: C07H 21/00 Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
• CPC Class Codes: C07K 1/04 on carriers C07K1/003, C07K1/006 take precedence
• CPC Class Codes: C07K 1/042 : characterised by the nature of the carrier
• CPC Class Codes: C40B 40/06 : Libraries containing nucleotides or polynucleotides, or derivatives thereof
• CPC Class Codes: C40B 40/10 : Libraries containing peptides or polypeptides, or derivatives thereof
• CPC Class Codes: C40B 60/14 : for creating libraries
• CPC Class Codes: G03F 7/00 : Photomechanical, e.g. photolithographic, production of textured or patterned surfaces, e.g. printing surfaces; Materials therefor, e.g. comprising photoresists; Apparatus specially adapted therefor using photoresist structures for special production processes, see the relevant places, e.g. B44C, H01L, e.g. H01L21/00, H05K
• CPC Class Codes: Y02P 20/55 : Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups

Abstract

A method for cyclization and reversal of the polarity of polymers on a substrate. The method provides for the formation of a polymer on a substrate (2) with a tether molecule (4). Through unmasking of a protective group (PG₂) a cyclic polymer (6) is formed. Through cleavage of an appropriate bond, a polarity reversed polymer (8) is formed. The method finds particular application in the formation of, for example, peptides and oligonucleotides.

Claims

1. A method of synthesizing a polymer on a substrate comprising the steps of:

a) on a surface of said substrate, having covalently attached thereto a molecule having the general formula;

##STR12## wherein said molecule is selected from the group consisting of;

##STR13## where;

X and Y are first and second reactive sites, respectively, X is selected from the group consisting of --O--, --NH--, --S--, --CO₂ --, --S--CH₂ CH₂ O--, and --NHCOCH₂ CH₂ CO₂ --; and

Y is selected from the group consisting of --OH, --NH₂, --SH, --CO₂ H, --S--CH₂ CH₂ OH, and NHCOCH₂ CH₂ CO₂ H, wherein Y is protected with a protective group PG₁ ;

T is a tether molecule;

PG₁ is a first protective group and is selected from the group consisting of BOC, FMOC, NVOC, DMT, NV, and FM;

PG₂ is a second protective group and is selected from the group consisting of BOC, FMOC, NVOC, DMT, NV, and FM; and

R and R'"'"' are independently selected from the group consisting of hydrogen, methyl, phenyl, and methoxyphenyl;

b) covalently coupling a polymer to said reactive site Y by;

removing said first protective group PG₁ from said reactive site Y;

covalently coupling a monomer to said reactive site Y, said first monomer having a monomer reactive site Y protected with a first protective group PG₁, wherein said first protective group PG₁ is selected from the group consisting of BOC, FMOC, NVOC, DMT, NV, and FM;

repeating said steps of removing a protective group and coupling a monomer to form a polymer at said reactive site Y;

c) removing said second protective group PG₂ ; and

d) covalently coupling a region of said polymer to said tether at said first reactive site to form a cyclic polymer.

2. A method as recited in claim 1 further comprising the step of cleaving said bond between T and Y whereby a polarity of said polymer is reversed.

3. The method as recited in claim 2 wherein said step of cleaving comprises the step of exposing said polymer to an acid.

4. The method as recited in claim 1 wherein said polymer is a peptide.

5. The method as recited in claim 4 wherein said polymer is a peptide and said monomer reactive site is an amino group.

6. The method as recited in claim 5 further comprising the step of cleaving said bond between T and Y whereby a polarity of said peptide is reversed.

7. The method as recited in claim 1 wherein said polymer is an oligonucleotide.

8. The method as recited in claim 1 wherein said polymer is an oligonucleotide and said monomer reactive site is a 5'"'"' end of said oligonucleotide.

9. The method as recited in claim 8 further comprising the step of cleaving said bond between T and Y whereby a polarity of said oligonucleotide is reversed.

10. The method as recited in claim 1 wherein said first protective group is NVOC and said second protective group is FMOC.

11. The method as recited in claim 1 wherein said first protective group is NVOC and said second protective group is BOC.

12. The method as recited in claim 1 wherein said first protective group is NVOC and said second protective group is DMT.

13. The method as recited in claim 1 wherein said first protective group is FMOC and said second protective group is NVOC.

14. The method as recited in claim 1 wherein said first protective group is DMT and said second protective group is NVOC.

15. The method as recited in claim 1 wherein said first protective group is NVOC and said second protective group is DMT, and said polymer is an oligonucleotide.

16. A method as recited in claim 1 wherein said molecule having the general formula:

##STR14## is a molecule having the general formula;

##STR15## where;

X and Y are first and second reactive sites, respectively; and

PG₁ and PG₂ are first and second protecting groups, respectively.

17. A method as recited in claim 1 wherein said molecule having the general formula:

##STR16## is a molecule having the general formula;

##STR17## where;

X and Y are first and second reactive sites, respectively;

PG₁ and PG₂ are first and second protecting groups, respectively; and

R and R'"'"' are independently selected from the group consisting of hydrogen, methyl, phenyl, and methoxyphenyl.

18. A method as recited in claim 17 wherein said molecule having the general formula:

##STR18## is a molecule having the general formula;

##STR19## where;

X and Y are first and second reactive sites, respectively;

PG₁ and PG₂ are first and second protecting groups, respectively; and

R and R'"'"' are independently selected from the group consisting of hydrogen, methyl, phenyl, and methoxyphenyl.

19. A method as recited in claim 1 wherein said molecule having the general formula:

##STR20## is a molecule having the general formula;

##STR21## where;

X and Y are first and second reactive sites, respectively;

PG₁ and PG₂ are first and second protecting groups, respectively; and

R and R'"'"' are independently selected from the group consisting of hydrogen, methyl, phenyl, and methoxyphenyl.

20. A method as recited in claim 19 wherein said molecule having the general formula:

##STR22## is a molecule having the general formula;

##STR23## where;

X and Y are first and second reactive sites, respectively;

PG₁ and PG₂ are first and second protecting groups, respectively; and

R and R'"'"' are independently selected from the group consisting of hydrogen, methyl, phenyl, and methoxyphenyl.

21. A method as recited in of claim 1 wherein said first protective group is DMT or BOC and said second protective group is FMOC.

22. A method as recited in claim 1 wherein said first protective group is selected from the group consisting of DMT, BOC, and FMOC and said second protective group is selected from the group consisting of NVOC, dimethoxybenzyl, dimethoxydimethylbenzyl, α

-methylphenoxycinnamates, nitroveratryl, and nitropiperonyl.

23. A method as recited in claim 1 wherein said first protective group is selected from the group consisting of NVOC, dimethoxybenzyl, dimethoxydimethylbenzyl, α

-methylphenoxycinnamates, nitroveratryl, and nitropiperonyl and said second protective group is selected from the group consisting of DMT, BOC, and FMOC.

24. A method of synthesizing an oligopeptide on a substrate comprising the steps of:

a) on a surface of said substrate, having covalently attached thereto a molecule having the general formula;

##STR24## where;

R and R'"'"' are independently selected from the group or hydrogen, C₂ -C₈ alkyl, phenyl, and methoxyphenyl;

Y is a reactive site selected from the group consisting of --OH, --NH₂, --SH, --CO₂ H, --S--CH₂ CH₂ OH, and NHCOCH₂ CH₂ CO₂ H and wherein Y is protected with a protective group PG₁ and;

A is oxygen, nitrogen, sulfur, CO, or CO₂ ;

n is 1 to 10;

PG₁ is a first protective group and is selected from the group consisting of BOC, FMOC, NVOC, DMT, NV, and FM; and

PG₂ is a second protective group and is selected from the group consisting of BOC, FMOC, NVOC, DMT, NV, and FM;

b) repeating the steps of;

1) removing said protective group PG₁ to provide a first reactive site Y;

2) covalently coupling an amino acid to said first reactive site Y, said first amino acid having said reactive site Y protected with said protective group PG₁ ;

to form an oligopeptide at said first reactive site Y;

c) removing said protective group PG₂ to provide a second reactive site; and

d) covalently coupling a region of said oligopeptide to said substrate bound molecule at said second reactive site to form a cyclic oligopeptide.