US 20060275801A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2006/0275801 A1 Henkin (43) Pub. Date: Dec. 7, 2006 (54) METHODS FOR DETECTION OF (52) U.S. Cl. .................................................. 435/6; 435/7.1 BOLOGICAL SUBSTANCES (76) Inventor: Robert I. Henkin, Bethesda, MD (US) (57) ABSTRACT Correspondence Address: WILSON SONSN GOODRCH & ROSAT The invention is directed to a method of detecting a bio 650 PAGE MILL ROAD logical Substance in the nasal Secretion and diagnosing a PALO ALTO, CA 94304-1050 (US) disease following the detection of the biological substance (21) Appl. No.: 11/415,942 wherein the biological Substance is not related to a respira tory disease. The invention also provides treatment of the (22) Filed: May 1, 2006 diseases following the detection of the biological Substance and/or diagnosis of the disease. In some embodiments, the Related U.S. Application Data diseases are cancer, hepatitis, Smell loss, taste loss, diabetes, (60) Provisional application No. 60/743,495, filed on Mar. and leprosy. The invention also provides a kit for diagnosing 15, 2006. Provisional application No. 60/676,252, a disease. The present invention includes methods of ana filed on Apr. 29, 2005. lyzing samples from the nose for the detection of biological Substances. In particular, nasal Secretion or nasal mucus is Publication Classification collected and analyzed for biological substances. The results (51) Int. Cl. of this analysis are then Suitable for use in diagnosis, CI2O I/68 (2006.01) prognosis, and determination of Suitability of therapeutic GOIN 33/53 (2006.01) interventions. Patent Application Publication Dec. 7, 2006 Sheet 1 of 12 US 2006/0275801 A1 3.Se3s?p9?,3soufield Patent Application Publication Dec. 7, 2006 Sheet 2 of 12 US 2006/0275801 A1 Z'50||-|| –002 Patent Application Publication Dec. 7, 2006 Sheet 3 of 12 US 2006/0275801 A1 Patent Application Publication Dec. 7, 2006 Sheet 4 of 12 US 2006/0275801 A1 L anes 1-7 HLA M Mol, wt markers 8-14 B-globin FIG. 4 Patent Application Publication Dec. 7, 2006 Sheet 5 of 12 US 2006/0275801 A1 Lp/(-)p-eoueose On Patent Application Publication Dec. 7, 2006 Sheet 6 of 12 US 2006/0275801 A1 P Name Calculate. CrOS. 1 WB POS COnt O.OOOE-00 22.67 2 R. BIk CO.OOOE-00 D41.00 3Specimen it O.OOOE-00 19.02 4 Specimen #2 O.OOOE-00 27.42 5 Speciment:1 + Pos Cont 0.000E+00 18.18 6 Specimen #1 + Pos Cont 0.000E+00 18.62 7 RBIk CO.OOOE-00 D4100 3.5 d2. 0.5O 242 12th- -n - is a < - -0.5 Cycle Number F.G. 6 Patent Application Publication Dec. 7, 2006 Sheet 7 of 12 US 2006/0275801 A1 Patent Application Publication Dec. 7, 2006 Sheet 8 of 12 US 2006/0275801 A1 8 WB POSCOnt 00 9 R. BIK . 0.000E-00 40.28 10 Specimen #1 0.000E+00 28.85 11 Specimen #2 12 Specimen #1 + Pos Cont 0.000E+00 22.13 13 Specimen #1 + Pos Cont 14 R. BIk 2. 5 0.1 -0. 5 Cycle Number FIG. 8 Patent Application Publication Dec. 7, 2006 Sheet 9 of 12 US 2006/0275801 A1 P Name Calculate. CrOS. 1 WBPOS COnt 0.000E+00 22.67 2 RBIK <0.000E+00 D41.00 3 Specimen #1 0.000E--00 19.02 4 Specimen #2 0.000E+00 27.42 5 Specimen #1+Pos Cont 0.000E+00 18.18 6 Specimen #1 + Pos Cont, 0.000E+00 18.62 7 RBIK <0.000E+00 D41.00 8 WBPOS COnt 0.000E+00 24.24 9 RBIK 0.000E+00 40.28 10 Specimen #1 0.000E+00 28.85 11 Specimen #2 12 Speciment:1 + Pos Cont 0.000E+00 22.13 13 Specimen #1 + Pos Cont 14 R. BK 13 12 11 10 9 8 7 3 6 L 5 4 3. 2 1 O O -1 -2 | | | | | | | O 5 10 15 20 25 30 35 40 45 Cycle Number FIG. 9 Patent Application Publication Dec. 7, 2006 Sheet 10 of 12 US 2006/0275801 A1 : 4 SWOLI19?jepueQuojºqeA??esp?oueaJoSISATIVNVSWHOL-ICIT?S :-***************************)009090000G0000x 33.Te?O/ssew …….…••••••••****************Ö000£0000z –?????????????????????????????????????? An a.ISuelu eAlbe Patent Application Publication Dec. 7, 2006 Sheet 11 of 12 US 2006/0275801 A1 x'x?'IIA–BLSVI.TIBOIIIJdÅL-31SVL - }JO?VHBÌN35) TVILNEJ.Od NY/HOeo!-TIJN |-TEWSTIBOTV8-LIW–BTBWS HO_Ld303}} Patent Application Publication Dec. 7, 2006 Sheet 12 of 12 US 2006/0275801 A1 z?‘SH QuB.I0pO/Que?SEJL | US 2006/0275801 A1 Dec. 7, 2006 METHODS FOR DETECTION OF BIOLOGICAL 0007 One aspect of the present invention is method of SUBSTANCES treating a disease wherein the treatment is based on diag nosing the disease, wherein the diagnosis is based on a CROSS-REFERENCE detection of a biological Substance in a nasal specimen, and 0001. This application is related to and claims priority to wherein the biological Substance is not related to a respira U.S. Provisional Application No. 60/743,495, filed Mar. 15, tory disease. In some embodiments, the disease is selected 2006, and U.S. Provisional Application No. 60/676.252, from the group consisting of Smell loss, taste loss, diabetes, filed Apr. 29, 2005, which are incorporated herein by ref obesity, anorexia, cancer, leprosy, and hepatitis. In some erence in their entirety. preferred embodiments, the cancer is ovarian cancer. In Some embodiments, the treatment is selected from the group BACKGROUND OF THE INVENTION consisting of oral administration, transmucosal administra tion, buccal administration, nasal administration, parental 0002 Detection and identification of biological sub administration, intravenous, Subcutaneous, intramuscular, stances in tissue samples is used for the diagnosis, progno Sublingual, transdermal administration, and rectal adminis sis, and monitoring of diseases. Efficient identification of tration. In some preferred embodiments, the treatment is by biological Substances aids in devising effective treatment nasal administration. Further the efficacy of the treatement Strategies. can be monitored by analysis of the nasal specimens. 0003 Most of the current diagnostic techniques involve 0008 Another aspect of the present invention is a method invasive procedures for the removal of tissue samples. of providing a conclusion regarding a disease to a patient, a Hence, there is need for the development of minimally health care provider or a health care manager where the invasive procedures for biological sample retrieval. conclusion is based on a diagnosis, wherein the diagnosis is 0004 The present invention provides methods for detec based on a detection of a biological Substance in a nasal tion of biological Substances, diagnosis of diseases based on specimen, and wherein the biological Substance is not this detection and methods for treatment of the diseases after related to a respiratory disease. the diagnosis. 0009. One aspect of the present invention is a method of diagnosing diabetes in a patient by obtaining a nasal speci SUMMARY OF THE INVENTION men, detecting insulin, insulin receptor or a combination 0005 One aspect of the present invention is a method of thereof in the specimen, and diagnosing the diabetes in the diagnosing a disease by obtaining a nasal specimen, detect patient, wherein the diagnosis is based on the detection of ing a biological Substance in the specimen, and diagnosing insulin, insulin receptor or a combination thereof in the nasal the disease, wherein the diagnosis is based on the detection specimen. Another aspect of the present invention is a of the biological Substance, and wherein the biological method of monitoring an efficacy of a diabetes therapy in a Substance is not related to a respiratory disease. In some patient by obtaining a nasal specimen, detecting insulin, embodiments, the biological Substance is a nucleic acid. In insulin receptor or a combination thereof in the specimen, some preferred embodiments, the nucleic acid is DNA, RNA and determining the efficacy of the diabetes therapy, wherein or a combination thereof. In some embodiments, the detec the determining is based on the detection of the insulin, tion is by a nucleic acid detection method. In some embodi insulin receptor or a combination thereof in the nasal speci ments, the nucleic acid detection method is selected from the C. group consisting of polymerase chain reaction (PCR), ligase 0010. One aspect of the present invention is a method of chain reaction (LCR), Strand displacement amplification diagnosing cancer in a patient by obtaining a nasal speci (SDA), self-sustained sequence replication (3SR), array men, detecting TNFC. in the specimen, and diagnosing the based test, and TAOMAN. In some preferred embodiments, cancer in the patient, wherein the diagnosis is based on the the nucleic acid detection method is PCR. detection of TNFC. in the nasal specimen. Another aspect of 0006. In some embodiments of the present invention, the the present invention is a method of monitoring an efficacy biological Substance is selected from the group consisting of of a cancer therapy by obtaining a nasal specimen, detecting insulin, insulin receptor, leptin, agouti-related protein, ghre TNFC. in the specimen, and determining the efficacy of the lin, glucose, growth factor, caspase, adenylyl cyclase, and cancer therapy, wherein the determining is based on the carbonic anhydrase. In some embodiments, the biological detection of TNFC. in the nasal specimen. Yet another aspect Substance is selected from the group consisting of p53, or of the present invention is a method of treating cancer by mutated p53, TNFO, TNFR I, TNFR II, TRAIL, IL3, obtaining a nasal specimen, detecting TNFC. in the speci endostatin, erythropoietin, bone morphogenic protein, brain men, and administering a drug to the patient, wherein the derived neurotrophic factor, ciliary neurotrophic factor, drug modulates the TNFC.