You’re using a public version of DrugPatentWatch with 5 free searches available | Register to unlock more free searches. CREATE FREE ACCOUNT

Last Updated: March 29, 2024

Claims for Patent: 4,801,531


✉ Email this page to a colleague

« Back to Dashboard


Summary for Patent: 4,801,531
Title: Apo AI/CIII genomic polymorphisms predictive of atherosclerosis
Abstract:The invention offers an early detection method for atherosclerosis using genetic analysis to detect a polymorphisms shown to be correlated with this disease which are proximal to the apolipoprotein AI (apoAI) and aplipoprotein CIII (apoCIII) gene complex. All individuals with a 300 bp deletion 4 kb upstream of the apoAI gene are destined to experience severe atherosclerotic symptomologies. Individuals with a polymorphism 5.4 kb 5\' of the apoAI gene or a PvuII polymorphism in the first intron of the apoCIII gene also seem to be at greater risk. A haplotype with MspI and XmnI/7.2 polymorphisms in this general region seem to be protected. Additional polymorphic sites in the DNA sequence associated with the apoAI/CIII gene complex provide a means for genetically fingerprinting individuals, and for identifying persons at risk with respect to disorders relating to lipid metabolism and transport.
Inventor(s): Frossard; Philippe M. (Palo Alto, CA)
Assignee: Biotechnology Research Partners, Ltd. (Mountain View, CA)
Application Number:06/782,666
Patent Claims:1. A method for predicting the development of atherosclerosis in an individual human subject, which method comprises detecting the presence or absence of the "XmnI/8.2" polymorphism, which is a deletion of a 300 bp segment of DNA at 4 kb 5' of the apoAI gene.

2. The method of claim 1 which comprises detecting the presence of absence of a diagnostic length DNA fragment of human genomic DNA which has been digested with a restriction endonuclease, said fragment hybridizing to p5'AI probe, or to a substantially equivalent probe.

3. The method of claim 2 wherein the restriction endonuclease is XmnI and the length of the fragment is 8.2 kb.

4. The method of claim 2 wherein the restriction endonuclease is ApaI and the length of the restriction fragment is 3.2 kb.

5. The method of claim 2 wherein the restriction endonuclease is BglI and the length of the fragment is 5.9 kb.

6. The method of claim 2 wherein the restriction endonuclease is BstEII and the length of the fragment is 2.1 kb.

7. The method of claim 2 wherein the restriction endonuclease is MspI and the length of the fragment is 5.9 kb.

8. The method of claim 2 wherein the restriction endonuclease is PvuII and the length of the fragment is 1.6 kb.

9. The method of claim 2 wherein the restriction endonuclease is RsaI and the length of the fragment is 3.2 kb.

10. The method of claim 2 wherein the restriction endonuclease is StuI and the length of the fragment is 5.5 kb.

11. The method of claim 2 wherein the restriction endonuclease is TaqI and the length of the fragment is 9.1 kb.

12. A method for predicting the development of atherosclerosis in an individual human, which method comprises detecting the presence or absence of the "ApaI" polymorphism which is 5.4 kb 5' of the apoAI gene.

13. The method of claim 12 which comprises detecting the presence of absence of a 2.2 kb DNA fragment of human genome which has been digested with ApaI, said fragment hybridizing with p5'AI probe or a substantially equivalent probe.

14. A method for predicting the development of atherosclerosis in an individual human, which method comprises detecting the presence of absence of the "PvuII" polymorphism which is in the first intron of the apoCIII gene.

15. The method of claim 15 which comprises detecting the presence or absence of a 0.87 kb DNA fragment of human genome which has been digested with PvuII, said fragment hybridizing with apoCIII probe or a substantially equivalent probe.

16. A method for predicting decreased susceptibility to atherosclerosis in an individual human which method comprises detecting the presence or absence of the "XmnI/7.2" polymorphism which is 3.7 kb 5' of the apoAI gene and the "MspI" polymorphism which is in the third intron of the apoAI gene.

17. The method of claim 16 which comprises detecting the presence or absence of a 7.2 kb DNA fragment of human genome which has been digested with XmnI, said fragment hybridizing with apoAI or p5'AI probe or a substantially equivalent probe and a 1.75 kb DNA fragment of human genome digested with MspI, said fragment hybridizing with apoAI probe or a substantially equivalent probe.

18. A method for ascertaining individual human subjects having a propensity for disorders relating to lipid transport and metabolism, which method comprises detecting the presence or absence of one or more polymorphisms selected from the group consisting of:

the "ApaI" polymorphism 5.4 kb 5' of the apoAI gene;

the "XmnI/8.2" polymorphism 4 kb 5' of the apoAI gene;

the "XmnI/7.2" polymorphism 3.7 kb 5' of the apoAI gene in combination with the "MspI" polymorphism in the 3rd intron of the apoAI gene; and

the "PvuII" polymorphism in the 1st intron of the apoCIII gene.

19. The method of claim 18 wherein each of said polymorphisms is detected in total human genomic DNA by, respectively,

digestion with ApaI and detection of a 2.2 kb fragment using p5'AI probe or a substantial equivalent;

digestion with XmnI and detection of an 8.2 kb fragment using p5'AI or a substantial equivalent or apoAI or a substantial equivalent;

digestion with XmnI and detection of a 7.2 kb fragment using p5'AI or a substantial equivalent or apoAI or a substantial equivalent in combination with digestion with MspI and detection of 1.75 kb fragment using apoAI probe or a substantial equivalent; and

digestion with PvuII and detection of a 0.87 kb fragment using apoCIII probe or a substantial equivalent.

20. A method of genetically identifying an individual, which method comprises assessing the presence or absence of at least one polymorphism by

contacting a genomic DNA digest with apoAI probe or a substantial equivalent, wherein said digest is obtained by digesting the genome with an enzyme selected from the group consisting of XmnI, HgiAI, and PstI, or a subset thereof, and

detecting the presence or absence of a diagnostic length DNA fragment which hybridizes to said probe.

21. A method of genetically identifying an individual, which method comprises assessing the presence or absence of at least one polymorphism by

contacting a genomic DNA digest with p5'AI probe or a substantial equivalent, wherein said digest is obtained by digesting the genome with an enzyme selected from the group consisting of ApaI, XmnI, BglI, BstEII, PvuII, RsaI, StuI and TaqI or a subset thereof, and

detecting the presence of absence of a diagnostic length DNA fragment which hybridizes to said probe.

22. A method of genetically identifying an individual which method comprises assessing the presence or absence of at least one polymorphism by

contacting a genomic DNA digest with apoCIII probe or a substantial equivalent wherein said digest is obtained by digesting the genome with BanII, PvuII or both.

23. The method of claim 2 wherein the restriction endonuclease is selected from the group consisting of XmnI, ApaI, BglI, BstEII, MspI, PvuII, RsaI, StuI, and TagI and the length of the fragment is, respectively, 8.2 kb, 3.2 kb, 5.9 kb, 2.1 kb, 5.9 kb, 1.6 kb, 3.2 kb, 5.5 kb, 9.1 kb.

24. A method of genetically identifying an individual, which method comprises detecting the presence or absence of one or more polymorphisms selected from the group consisting of:

the "ApaI" polymorphism 5.4 kb 5' of the apoAI gene;

the "XmnI/8.2" polymorphism 4 kb 5' of the apoAI gene;

the "XmnI/7.2" polymorphism 3.7 kb 5' of the apoAI gene in combination with the "MspI" polymorphism in the 3rd intron of the apoAI gene;

the "HgiAI" polymorphism in the 3rd exon of the apoAI gene;

the "PstI" polymorphism 0.3 kb 3' of the apoAI gene; and

the "PvuII" polymorphism in the 1st intron of the apoCIII gene.

25. The method of claim 24 wherein each said polymorphism is detected in total human genomic DNA by, respectively,

digestion with ApaI and detection of a 2.2 kb fragment using p5'AI probe or a substantial equivalent;

digestion with XmnI and detection of an 8.2 kb fragment using p5'AI or a substantial equivalent or apoAI or a substantial equivalent;

digestion with XmnI and detection of a 7.2 kb fragment using p5'AI or a substantial equivalent or apoAI or a substantial equivalent

in combination with digestion with MspI and detection of 1.75 kb fragment using apoAI probe or a substantial equivalent;

digestion with HgiAI and detection of 0.94 kb fragment using apoAI probe or a substantial equivalent;

digestion with PstI detection of a 3.2 kb fragment using apoAI probe or a substantial equivalent; and

digestion with PvuII and detection of a 0.87 kb fragment using apoCIII probe or a substantial equivalent.

26. A method to detect the SstI/BanII polymorphism in the 4th exon of the gene which comprises digesting the genome with BanII and detecting a 1.0 and 0.8 kb fragment using apoCIII probe or a substantial equivalent.

27. A reagent kit useful in performing the method of claim 1, which kit includes XmnI restriction enzyme and at least one probe selected from apoAI gene probe or a substantial equivalent and p5'AI probe or a substantial equivalent.

28. A reagent kit useful in performing the method of claim 1, which kit includes p5'AI probe or substantial equivalent and at least one restriction enzyme selected from the group consisting of ApaI, XmnI, BglI, BstEII, MspI, PvuII, RsaI, StuI, and TaqI.

29. A reagent kit useful in performing the method of claim 12, which kit includes ApaI restriction enzyme and p5'AI probe or a substantial equivalent.

30. A reagent kit useful in performing the method of claim 14, which kit includes PvuII restriction enzyme and apoCIII probe or a substantial equivalent.

31. A reagent kit useful in performing the method of claim 16, which kit includes XmnI restriction enzyme MspI restriction enzyme, and apoAI probe or a substantial equivalent.

32. A reagent kit useful in ascertaining individual human subjects having a propensity for disorders related to lipid transport and metabolism, which kit includes at least one probe selected from the group consisting of apoAI probe, p5'AI probe, and apoCIII probe, or their substantial equivalents, and at least one restriction enzyme selected from the group consisting of ApaI, XmnI, HgiAI, MspI, BanII, PvuII, BglI, BstEII, RsaI, StuI, and TaqI.

Details for Patent 4,801,531

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 ⤷  Try a Trial 2039-02-26
Merck Sharp & Dohme Corp. INTRON A interferon alfa-2b For Injection 103132 ⤷  Try a Trial 2039-02-26
Merck Sharp & Dohme Corp. INTRON A interferon alfa-2b Injection 103132 ⤷  Try a Trial 2039-02-26
>Applicant >Tradename >Biologic Ingredient >Dosage Form >BLA >Approval Date >Patent No. >Expiredate

Make Better Decisions: Try a trial or see plans & pricing

Drugs may be covered by multiple patents or regulatory protections. All trademarks and applicant names are the property of their respective owners or licensors. Although great care is taken in the proper and correct provision of this service, thinkBiotech LLC does not accept any responsibility for possible consequences of errors or omissions in the provided data. The data presented herein is for information purposes only. There is no warranty that the data contained herein is error free. thinkBiotech performs no independent verification of facts as provided by public sources nor are attempts made to provide legal or investing advice. Any reliance on data provided herein is done solely at the discretion of the user. Users of this service are advised to seek professional advice and independent confirmation before considering acting on any of the provided information. thinkBiotech LLC reserves the right to amend, extend or withdraw any part or all of the offered service without notice.