US5139940A — Activation compounds and methods of synthesis of …

Claim 1

Independent Claim 61 words

A compound having the formula: ##STR18## in which R 1 is --OH, --Br, --Cl, --I, --NH 2 , or --NH 3 + X - , where X - is Br - , Cl - or I - , and where R 2 , R 3 and R 4 , which may be the same or different, are --H or 3 H.

Claim 2

Dependent Claim 61 words

A compound having the formula: ##STR18## in which R 1 is --OH, --Br, --Cl, --I, --NH 2 , or --NH 3 + X - , where X - is Br - , Cl - or I - , and where R 2 , R 3 and R 4 , which may be the same or different, are --H or 3 H.

Claim 3

Dependent Claim 34 words

A compound having the formula: ##STR19## in which R 1 is --Br or --I, and where R 2 and R 3 , which may be the same or different, are --H or 3 H.

Claim 4

Dependent Claim 34 words

A compound having the formula: ##STR19## in which R 1 is --Br or --I, and where R 2 and R 3 , which may be the same or different, are --H or 3 H.

Claim 5

Dependent Claim 82 words

A compound having the formula: ##STR20## in which R 1 is ##STR21## where n is a whole number between 6 and 16, where R 2 , R 3 and R 4 , in which may be the same or different, are --H or 3 H, and where R 5 =R 7 =--CH 3 , --C 2 H 5 , --C 3 H 7 or --C 4 H 9 and R 6 may have the formula --H or: ##STR22## or a salt thereof.

Claim 6

Dependent Claim 82 words

A compound having the formula: ##STR20## in which R 1 is ##STR21## where n is a whole number between 6 and 16, where R 2 , R 3 and R 4 , in which may be the same or different, are --H or 3 H, and where R 5 =R 7 =--CH 3 , --C 2 H 5 , --C 3 H 7 or --C 4 H 9 and R 6 may have the formula --H or: ##STR22## or a salt thereof.

Claim 7

Dependent Claim 78 words

A compound having the formula: ##STR23## in which R 1 is ##STR24## where n is a whole number between 6 and 16, where R 2 and R 3 , which may be the same or different, are --H or 3 H, and where R 4 =R 6 =--CH 3 , --C 2 H 5 , --C 3 H 7 or --C 4 H 9 and R 5 may have the formula --H or: ##STR25## or a salt thereof.

Claim 8

Dependent Claim 78 words

A compound having the formula: ##STR23## in which R 1 is ##STR24## where n is a whole number between 6 and 16, where R 2 and R 3 , which may be the same or different, are --H or 3 H, and where R 4 =R 6 =--CH 3 , --C 2 H 5 , --C 3 H 7 or --C 4 H 9 and R 5 may have the formula --H or: ##STR25## or a salt thereof.

Claim 9

Dependent Claim 26 words

A compound having the formula: ##STR26## in which R 1 and R 2 , which may be the same or different, are --H or 3 H.

Claim 10

Dependent Claim 26 words

A compound having the formula: ##STR26## in which R 1 and R 2 , which may be the same or different, are --H or 3 H.

Claim 11

Dependent Claim 40 words

A compound having the formula: ##STR27## in which R 1 and R 2 , which may be the same or different, are --CH 3 , --C 2 H 5 , --C 3 H 7 or --C 4 H 9 .

Claim 12

Dependent Claim 40 words

A compound having the formula: ##STR27## in which R 1 and R 2 , which may be the same or different, are --CH 3 , --C 2 H 5 , --C 3 H 7 or --C 4 H 9 .

Claim 13

Dependent Claim 14 words

A compound having the formula: ##STR28## in which X is --Cl, --Br or --I.

Claim 14

Dependent Claim 14 words

A compound having the formula: ##STR28## in which X is --Cl, --Br or --I.

Claim 15

Dependent Claim 35 words

A compound having the formula: ##STR29## in which X is --Cl, --Br or --I and R is --CH 3 , --C 2 H 5 , --C 3 H 7 or --C 4 H 9 .

Claim 16

Dependent Claim 35 words

A compound having the formula: ##STR29## in which X is --Cl, --Br or --I and R is --CH 3 , --C 2 H 5 , --C 3 H 7 or --C 4 H 9 .

Claim 17

Dependent Claim 29 words

A compound having the formula: ##STR30## in which R 1 , R 2 and R 3 , which may be the same or different, are --H or 3 H.

Claim 18

Dependent Claim 29 words

A compound having the formula: ##STR30## in which R 1 , R 2 and R 3 , which may be the same or different, are --H or 3 H.

Claim 19

Dependent Claim 22 words

A complex, comprising one or more of the compounds of claims 1, 2, 3, 4, 5, or 9, coupled to nucleic acid.

Claim 20

Dependent Claim 11 words

A complex as in claim 10 wherein said coupling is non-covalent.

Claim 21

Dependent Claim 11 words

A complex as in claim 10 wherein said coupling is covalent.

Claim 22

Dependent Claim 24 words

A complex as in claim 10 wherein said nucleic acid is selected from the group consisting of viral, bacterial, fungal, mycoplasmal and protozoan DNA.

Claim 23

Dependent Claim 14 words

A complex as in claim 10 wherein said nucleic acid is human nucleic acid.

Claim 24

Dependent Claim 24 words

A complex, comprising 5-aminomethylisopsoralen coupled to nucleic acid at a level of addition greater than one 5-aminomethylisopsoralen molecule per fifty nucleic acid base pairs.

Claim 25

Dependent Claim 24 words

A complex, comprising 5-aminomethylisopsoralen coupled to nucleic acid at a level of addition greater than one 5-aminomethylisopsoralen molecule per fifty nucleic acid base pairs.

Claim 26

Dependent Claim 24 words

A complex, comprising 4'-aminomethyl-4,5'-dimethylisopsoralen coupled to nucleic acid at a level of addition greater than one 4'-aminomethyl-4,5'-dimethylisopsoralen molecule per fifty nucleic acid base pairs.

Claim 27

Dependent Claim 24 words

A complex, comprising 4'-aminomethyl-4,5'-dimethylisopsoralen coupled to nucleic acid at a level of addition greater than one 4'-aminomethyl-4,5'-dimethylisopsoralen molecule per fifty nucleic acid base pairs.

Claim 28

Dependent Claim 14 words

A complex as in claims 15 or 16 wherein said nucleic acid is DNA.

Claim 29

Dependent Claim 14 words

A complex as in claims 15 or 16 wherein said nucleic acid is RNA.

Claim 30

Independent Claim 24 words

A method of synthesizing 5-methylisopsoralen, comprising the steps: a) providing 7-hydroxy-5-methylcoumarin; and b) treating 7-hydroxy-5-methylcoumarin with a haloethylene carbonate so that 5-methylisopsoralen is produced.

Claim 31

Independent Claim 24 words

A method of synthesizing 5-methylisopsoralen, comprising the steps: a) providing 7-hydroxy-5-methylcoumarin; and b) treating 7-hydroxy-5-methylcoumarin with a haloethylene carbonate so that 5-methylisopsoralen is produced.

Claim 32

Independent Claim 34 words

A method of synthesizing 5-methylisopsoralen, comprising the steps: a) providing 7-hydroxy-5-methylcoumarin; b) treating 7-hydroxy-5-methylcoumarin with a haloacetaldehyde diethylacetal so that 7-(2,2-diethoxyethyloxy)-5-methylcoumarin is produced; and c) treating 7-(2,2-diethoxyethyloxy)-5-methylcoumarin with acid so that 5-methylisopsoralen is produced.

Claim 33

Independent Claim 34 words

A method of synthesizing 5-methylisopsoralen, comprising the steps: a) providing 7-hydroxy-5-methylcoumarin; b) treating 7-hydroxy-5-methylcoumarin with a haloacetaldehyde diethylacetal so that 7-(2,2-diethoxyethyloxy)-5-methylcoumarin is produced; and c) treating 7-(2,2-diethoxyethyloxy)-5-methylcoumarin with acid so that 5-methylisopsoralen is produced.

Claim 34

Independent Claim 36 words

A method of synthesizing 4,5'-dimethylisopsoralen, comprising the steps: a) treating 7-(β-haloallyloxy)-4-methylcoumarin with heat and an alkyl anhydride to yield the esterified Claisen product 7-alkylcarbonyloxy-8-(β-haloallyl)-4-methylcoumarin; and b) treating 7-alkylcarbonyloxy-8-(β-halo-allyl)-4-methylcoumarin with an acid so that 4,5'-dimethylisopsoralen is produced.

Claim 35

Independent Claim 36 words

A method of synthesizing 4,5'-dimethylisopsoralen, comprising the steps: a) treating 7-(β-haloallyloxy)-4-methylcoumarin with heat and an alkyl anhydride to yield the esterified Claisen product 7-alkylcarbonyloxy-8-(β-haloallyl)-4-methylcoumarin; and b) treating 7-alkylcarbonyloxy-8-(β-halo-allyl)-4-methylcoumarin with an acid so that 4,5'-dimethylisopsoralen is produced.

Claim 36

Independent Claim 59 words

A method of synthesizing 4'-aminomethyl-4,5'dimethylisopsoralen, comprising the steps: a) treating 7-alkylcarbonyloxy-8-(β-haloallyl)-4-methylcoumarin with an acid so that 4,5'-dimethylisopsoralen is produced; b) treating 4,5'-dimethylisopsoralen with a halomethyl methylether to yield a halomethyl derivative; c) treating the halomethyl derivative with an alkali salt of phthalimide to yield the corresponding phthalimidomethyl derivative; and d) treating the phthalimidomethyl derivative with hydrazine to yield 4'-aminomethyl-4,5'dimethylisopsoralen.

Claim 37

Independent Claim 59 words

A method of synthesizing 4'-aminomethyl-4,5'dimethylisopsoralen, comprising the steps: a) treating 7-alkylcarbonyloxy-8-(β-haloallyl)-4-methylcoumarin with an acid so that 4,5'-dimethylisopsoralen is produced; b) treating 4,5'-dimethylisopsoralen with a halomethyl methylether to yield a halomethyl derivative; c) treating the halomethyl derivative with an alkali salt of phthalimide to yield the corresponding phthalimidomethyl derivative; and d) treating the phthalimidomethyl derivative with hydrazine to yield 4'-aminomethyl-4,5'dimethylisopsoralen.

Claim 38

Independent Claim 32 words

A method of synthesizing 4'-aminomethyl-4,5'dimethylisopsoralen, comprising the steps: a) providing 4,5'-dimethylisopsoralen; b) reacting 4,5'-dimethylisopsoralen with a nitrogen donor so that 4'-phthalimidomethyl-4,5'-dimethylisopsoralen is produced; and c) treating 4'-phthalimidomethyl-4,5'-dimethylisopsoralen with hydrazine to yield 4'-aminomethyl-4,5'dimethylisopsoralen.

Claim 39

Independent Claim 32 words

A method of synthesizing 4'-aminomethyl-4,5'dimethylisopsoralen, comprising the steps: a) providing 4,5'-dimethylisopsoralen; b) reacting 4,5'-dimethylisopsoralen with a nitrogen donor so that 4'-phthalimidomethyl-4,5'-dimethylisopsoralen is produced; and c) treating 4'-phthalimidomethyl-4,5'-dimethylisopsoralen with hydrazine to yield 4'-aminomethyl-4,5'dimethylisopsoralen.

Claim 40

Dependent Claim 24 words

The method of claim 23 wherein said nitrogen donor may be selected from the group consisting of phthalimide salts, N-hydroxymethyl phthalimide and hexamethylene tetramine.

Claim 41

Independent Claim 48 words

A method of synthesizing 4'-aminomethyl-4,5'dimethylisopsoralen, comprising the steps: a) providing 4'-hydroxymethyl-4,5'-dimethylisopsoralen; b) treating 4'-hydroxymethyl-4,5'-dimethylisopsoralen with a thionyl halide so that 4'-halomethyl-4,5'-dimethylisopsoralen is produced; c) treating 4'-halomethyl-4,5'-dimethylisopsoralen with an alkali salt of phthalimide so that 4'-phthalimidomethyl-4,5'-dimethylisopsoralen is produced; and d) treating 4'-phthalimidomethyl-4,5'-dimethylisopsoralen with hydrazine so that 4'-aminomethyl-4,5'-dimethylisopsoralen is produced.

Claim 42

Independent Claim 48 words

A method of synthesizing 4'-aminomethyl-4,5'dimethylisopsoralen, comprising the steps: a) providing 4'-hydroxymethyl-4,5'-dimethylisopsoralen; b) treating 4'-hydroxymethyl-4,5'-dimethylisopsoralen with a thionyl halide so that 4'-halomethyl-4,5'-dimethylisopsoralen is produced; c) treating 4'-halomethyl-4,5'-dimethylisopsoralen with an alkali salt of phthalimide so that 4'-phthalimidomethyl-4,5'-dimethylisopsoralen is produced; and d) treating 4'-phthalimidomethyl-4,5'-dimethylisopsoralen with hydrazine so that 4'-aminomethyl-4,5'-dimethylisopsoralen is produced.

Claim 43

Independent Claim 36 words

A method of synthesizing 5-aminomethyl-isopsoralen, comprising the steps: a) providing a 5-halomethylisopsoralen; b) treating said 5-halomethylisopsoralen with hexamethylenetetramine to create a hexamethylene-tetramine derivative; and c) treating said hexamethylenetetramine derivative with acid so that 5-aminomethylisopsoralen is produced.

Claim 44

Independent Claim 36 words

A method of synthesizing 5-aminomethyl-isopsoralen, comprising the steps: a) providing a 5-halomethylisopsoralen; b) treating said 5-halomethylisopsoralen with hexamethylenetetramine to create a hexamethylene-tetramine derivative; and c) treating said hexamethylenetetramine derivative with acid so that 5-aminomethylisopsoralen is produced.

Claim 45

Dependent Claim 16 words

The method of claim 26, wherein said providing of a 5-halomethylisopsoralen comprises synthesizing 5-bromomethylisopsoralen from 5-methylisopsoralen.

Claim 46

Independent Claim 65 words

A method of synthesizing radiolabelled 5-aminomethylisopsoralen, comprising the steps: a) providing formylisopsoralen; b) treating said formylisopsoralen with radillabelled sodium borohydride to yield the radiolabelled hydroxymethylisopsoralen; c) treating hydroxymethylisopsoralen with a thionyl halide to yield the corresponding radiolabelled halomethylisopsoralen; d) treating said halomethylisopsoralen with hexamethylene tetramine to create the corresponding radiolabelled hexamethylenetetramine derivative; and e) treating said hexamethylenetetramine derivative with an acid to produce radiolabelled 5-aminomethylisopsoralen.

Claim 47

Independent Claim 65 words

A method of synthesizing radiolabelled 5-aminomethylisopsoralen, comprising the steps: a) providing formylisopsoralen; b) treating said formylisopsoralen with radillabelled sodium borohydride to yield the radiolabelled hydroxymethylisopsoralen; c) treating hydroxymethylisopsoralen with a thionyl halide to yield the corresponding radiolabelled halomethylisopsoralen; d) treating said halomethylisopsoralen with hexamethylene tetramine to create the corresponding radiolabelled hexamethylenetetramine derivative; and e) treating said hexamethylenetetramine derivative with an acid to produce radiolabelled 5-aminomethylisopsoralen.

Claim 48

Independent Claim 28 words

A method for labelling nucleic acid, comprising the steps: a) providing labelled 5-aminomethylisopsoralen and nucleic acid; and b) coupling said labelled 5-aminomethylisopsoralen to said nucleic acid by photoactivation.

Claim 49

Independent Claim 28 words

A method for labelling nucleic acid, comprising the steps: a) providing labelled 5-aminomethylisopsoralen and nucleic acid; and b) coupling said labelled 5-aminomethylisopsoralen to said nucleic acid by photoactivation.

Claim 50

Independent Claim 45 words

A method for inhibiting template-dependent enzymatic synthesis of nucleic acid, comprising the steps: a) providing one or more furocoumarins and nucleic acid; and b) coupling said one or more furocoumarins to said nucleic acid by photoactivation, so that it cannot be replicated by Taq polymerase.

Claim 51

Independent Claim 45 words

A method for inhibiting template-dependent enzymatic synthesis of nucleic acid, comprising the steps: a) providing one or more furocoumarins and nucleic acid; and b) coupling said one or more furocoumarins to said nucleic acid by photoactivation, so that it cannot be replicated by Taq polymerase.

Claim 52

Independent Claim 45 words

A method for inhibiting template-dependent enzymatic synthesis of nucleic acid, comprising the steps: a) providing one or more furocoumarins and nucleic acid; and b) coupling said one or more furocoumarins to said nucleic acid by photoactivation, so that it cannot be replicated by T4 polymerase.

Claim 53

Independent Claim 45 words

A method for inhibiting template-dependent enzymatic synthesis of nucleic acid, comprising the steps: a) providing one or more furocoumarins and nucleic acid; and b) coupling said one or more furocoumarins to said nucleic acid by photoactivation, so that it cannot be replicated by T4 polymerase.

Claim 54

Independent Claim 26 words

The method as recited in claims 30 or 31 wherein said one or more furocoumarins are selected from the group consisting of 5-methylisopsoralen, 5-aminomethylisopsoralen and 4'-aminomethyl-4,5'-dimethylisopsoralen.

Claim 55

Independent Claim 45 words

A method for inhibiting template-dependent enzymatic synthesis of nucleic acid, comprising the steps: a) providing one or more isopsoralens and nucleic acid; and b) coupling said one or more isopsoralens to said nucleic acid by photoactivation, so that it cannot be replicated by reverse transcriptase.

Claim 56

Independent Claim 45 words

A method for inhibiting template-dependent enzymatic synthesis of nucleic acid, comprising the steps: a) providing one or more isopsoralens and nucleic acid; and b) coupling said one or more isopsoralens to said nucleic acid by photoactivation, so that it cannot be replicated by reverse transcriptase.

Claim 57

Dependent Claim 24 words

The method as recited in claim 33 wherein said one or more isopsoralens are selected from the group consisting of 5-methylisopsoralen, 5-aminomethylisopsoralen and 4'-aminomethyl-4,5'-dimethylisopsoralen.

Claim 58

Independent Claim 41 words

A method for inhibiting template-dependent enzymatic synthesis of nucleic acid, comprising the steps: a) providing 4'-aminomethyl-4,5'-dimethylisopsoralen and nucleic acid; and b) binding said 4'-aminomethyl-4,5'-dimethylisopsoralen to nucleic acid by photoactivation, so that it cannot be replicated by E. coli DNA polymerase I.

Claim 59

Independent Claim 41 words

A method for inhibiting template-dependent enzymatic synthesis of nucleic acid, comprising the steps: a) providing 4'-aminomethyl-4,5'-dimethylisopsoralen and nucleic acid; and b) binding said 4'-aminomethyl-4,5'-dimethylisopsoralen to nucleic acid by photoactivation, so that it cannot be replicated by E. coli DNA polymerase I.

Claim 60

Independent Claim 38 words

A method for inhibiting template-dependent enzymatic synthesis of nucleic acid, comprising the steps: a) providing 4'-aminomethyl-4,5'-dimethylisopsoralen and nucleic acid; and b) binding said 4'-aminomethyl-4,5'-dimethylisopsoralen to nucleic acid by photoactivation, so that it cannot be replicated by Klenow polymerase.

Claim 61

Independent Claim 38 words

A method for inhibiting template-dependent enzymatic synthesis of nucleic acid, comprising the steps: a) providing 4'-aminomethyl-4,5'-dimethylisopsoralen and nucleic acid; and b) binding said 4'-aminomethyl-4,5'-dimethylisopsoralen to nucleic acid by photoactivation, so that it cannot be replicated by Klenow polymerase.

Claim 62

Independent Claim 32 words

A method for modifying a target sequence, comprising the steps: a) providing nucleic acid containing a target sequence; b) providing 5-aminomethylisopsoralen; and c) coupling said 5-aminomethylisopsoralen to said target sequence by photoactivation.

Claim 63

Independent Claim 32 words

A method for modifying a target sequence, comprising the steps: a) providing nucleic acid containing a target sequence; b) providing 5-aminomethylisopsoralen; and c) coupling said 5-aminomethylisopsoralen to said target sequence by photoactivation.

Claim 64

Independent Claim 37 words

A method for detecting a target sequence, comprising the steps: a) providing nucleic acid containing a target sequence; b) providing 5-aminomethylisopsoralen; c) coupling said 5-aminomethylisopsoralen to said target sequence by photoactivation; and d) detecting said target sequence.

Claim 65

Independent Claim 37 words

A method for detecting a target sequence, comprising the steps: a) providing nucleic acid containing a target sequence; b) providing 5-aminomethylisopsoralen; c) coupling said 5-aminomethylisopsoralen to said target sequence by photoactivation; and d) detecting said target sequence.

Claim 66

Independent Claim 38 words

A method for detecting a target sequence, comprising the steps: a) providing nucleic acid containing a target sequence; b) providing 4'-aminomethyl-4,5'-dimethyl-isopsoralen; and c) coupling said 4'-aminomethyl-4,5'-dimethylisopsoralen to said target sequence by photoactivation; and d) detecting said target sequence.

Claim 67

Independent Claim 38 words

A method for detecting a target sequence, comprising the steps: a) providing nucleic acid containing a target sequence; b) providing 4'-aminomethyl-4,5'-dimethyl-isopsoralen; and c) coupling said 4'-aminomethyl-4,5'-dimethylisopsoralen to said target sequence by photoactivation; and d) detecting said target sequence.

Claim 68

Independent Claim 28 words

A method of synthesizing photoproduct, comprising the steps: a) providing one or more furocoumarins; and b) irradiating said one or more furocoumarins in the absence of nucleic acid.

Claim 69

Independent Claim 28 words

A method of synthesizing photoproduct, comprising the steps: a) providing one or more furocoumarins; and b) irradiating said one or more furocoumarins in the absence of nucleic acid.

Claim 70

Dependent Claim 23 words

The method as recited in claim 40 wherein said one or more furocoumarins are selected from the group consisting of 5-aminomethylisopsoralen and 4'-aminomethyl-4,5'-dimethylisopsoralen.

Claim 71

Independent Claim 57 words

A method of binding photoproduct to nucleic acid, comprising the sequential steps: a) providing in any order: i) one or more furocoumarin derivatives, and ii) nucleic acid; b) exposing said furocoumarin derivatives to activating wavelengths of electromagnetic radiation; c) mixing said irradiated furocoumarin derivatives with said nucleic acid in the absence of activating wavelengths of electromagnetic radiation.

Claim 72

Independent Claim 57 words

A method of binding photoproduct to nucleic acid, comprising the sequential steps: a) providing in any order: i) one or more furocoumarin derivatives, and ii) nucleic acid; b) exposing said furocoumarin derivatives to activating wavelengths of electromagnetic radiation; c) mixing said irradiated furocoumarin derivatives with said nucleic acid in the absence of activating wavelengths of electromagnetic radiation.

Annuity Stage	Window Opens	Payment Deadline	Surcharge Period Until	Estimated Fee	Status
3.5 Years (Year 4)	1995-08-18	1996-02-17	1996-08-18	$2000.00	Window Closed / Lapsed
7.5 Years (Year 8)	1999-08-18	2000-02-17	2000-08-18	$3760.00	Window Closed / Lapsed
11.5 Years (Year 12)	2003-08-18	2004-02-17	2004-08-18	$7700.00	Window Closed / Lapsed

Activation compounds and methods of synthesis of activation compounds

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