WO1998015835A1 - Procede de detection precoce du cancer des poumons par provocation de l'expectoration et analyse des crachats afin de detecter des substances associees au cancer - Google Patents

Procede de detection precoce du cancer des poumons par provocation de l'expectoration et analyse des crachats afin de detecter des substances associees au cancer Download PDF

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WO1998015835A1
WO1998015835A1 PCT/US1997/018137 US9718137W WO9815835A1 WO 1998015835 A1 WO1998015835 A1 WO 1998015835A1 US 9718137 W US9718137 W US 9718137W WO 9815835 A1 WO9815835 A1 WO 9815835A1
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sputum
lung cancer
utp
subject
compound
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PCT/US1997/018137
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WO1998015835A8 (fr
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Karol K. Lacroix
Christy L. Shaffer
Karla M. Jacobus
Janet L. Rideout
David J. Drutz
William Pendergast
Benjamin R. Yerxa
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Inspire Pharmaceuticals, Inc.
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Priority to AU49788/97A priority Critical patent/AU4978897A/en
Publication of WO1998015835A1 publication Critical patent/WO1998015835A1/fr
Publication of WO1998015835A8 publication Critical patent/WO1998015835A8/fr

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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
    • G01N33/57407—Specifically defined cancers
    • G01N33/57423—Specifically defined cancers of lung

Definitions

  • This invention relates to the diagnosis of cancer, specifically, to a method of sputum induction and cytologic assay for the early detection of lung cancer.
  • Lung cancer is the most common fatal malignant neoplasm of both men and women in the United States, with an approximate 13% to 15% survival rate 5 years after diagnosis (T. Petty, Med. Clin. North Amer. 80(3), 645-55 (1996)). Lung cancer typically occurs after a prolonged latent period of several years to several decades, during which the normal airway epithelium undergoes cellular changes which progressively become more severe until ultimately they progress to carcinoma in situ and to invasive carcinoma. However, if detected in its early stages, i.e., carcinoma in situ or microinvasive cancer, the potential for cure is essentially 100% (S. Lam, abstract presented at the Annual Respiratory Disease Symposium, Vancouver, B.C., October 27-28, 1995). Therapeutic measures in these early stages include photodynamic therapy, cryotherapy, or surgery. Premalignant lesions can be treated with chemopreventative agents such as 13- cis-retinoic acid to suppress and even reverse the carcinogenic process.
  • chemopreventative agents such as 13- cis-reti
  • the present invention discloses an effective means of early lung cancer detection by means of inducing expectoration of deep lung sputum with nucleoside phosphates, preferably uridine 5'-triphosphate (UTP) or P 1 , P 4 - di(uridine 5'-)tetraphosphate (U2P4), and then analyzing the sputum for the presence of labeled reagents selectively bound to substances in the sputum whose presence correlates with the development of lung cancer.
  • nucleoside phosphates preferably uridine 5'-triphosphate (UTP) or P 1 , P 4 - di(uridine 5'-)tetraphosphate (U2P4)
  • U2P4 uridine 5'-triphosphate
  • U2P4 uridine 5'-triphosphate
  • U2P4 uridine 5'-triphosphate
  • biomarkers of lung cancer can be, for example, cellular proteins whose enhanced presence correlates with the development of lung cancer, or
  • NCI National Cancer Institute
  • Mabs monoclonal antibodies
  • biom.arkers it is meant that the enhanced presence of these antigens on the surface of bronchial epithelial cells has been shown to be correlated with the later development of lung cancer.
  • a sputum specimen not originating from ⁇ deep in the lungs ⁇ are of little use because they typically do not contain exfoliated cells from the lung. Twenty percent of current smokers and 30% of former smokers do not produce a satisfactory sputum specimen for examination despite inhalation of hypertonic saline followed by a rigorous sputum induction procedure (S. Lam, supra).
  • the current invention discloses a method of facilitating induction of deep lung sputum for the purpose of early lung cancer detection.
  • researchers have also identified specific gene mutations which correlate with the subsequent development of lung cancer.
  • the first step in any lung cancer detection method based upon sputum analysis is the collection of an adequate sample of deep lung sputum.
  • the ability to expectorate sputum relies on the coordinated function of the mucociliary escalator and the cough mechanism.
  • the mucociliary escalator relies on the integrated action of three mech.anisms: 1) mucus section by goblet cells and submucosal glands; 2) cilia beating to propel the mucus out of the lungs; and 3) epithelial ion transport systems which maintain the ionic milieu of, and hence, the viscosity of airway surface liquid.
  • UTP has been shown to increase both the rate and total amount of mucin secretion by goblet cells in vitro (M. Lethem, et al., Am J. Respir. Cell Mol. BioL 9, 315-22 (1993)).
  • UTP has been shown to increase cilia beat frequency in human airway epithelial cells in vitro (D. Drutz, et al., Drug Development Research, 37(3), 185 (1996)).
  • UTP has been shown to increase intracellular Ca ++ concentration due to stimulation of phospholipase C as a result of initial occupation of the P 2 ⁇ 2 T receptor by UTP (H.
  • UTP's modulation of all three components of the mucociliary escalator system results in at least a 2.5-fold improvement in mucociliary clearance (MCC) without any significant side-effects in normal volunteers (as measured by radiolabel techniques and administered on an acute basis — long-term therapy has not yet been tested) (K. Olivier, et al, Am. J. Respir. Crit. Care Med. 154, 217-23 (1996)). It should be noted that in some disease states the cilia are not fully operational, e.g., primary ciliary dyskinesia, chronic bronchitis and the flu. UTP can still increase secretion clearance in patients with PCD via increased mucin production and hydration of the mucus (P.
  • hypertonic saline is usually administered by a specially trained respiratory therapist, and many smaller hospitals and clinics do not have these personnel on staff. Additionally, hypertonic saline is only effective about 70-80% of the time in inducing a sufficient sputum specimen from the lower lung (S. Lam, supra). Because of hypertonic saline ⁇ s shortcomings as a sputum induction agent, many hospitals no longer perform sputum cytology; instead they perform bronchoscopy in an attempt to locate and biopsy the neoplasm. Bronchoscopy is a much more expensive procedure, averaging about $2,000 per procedure, including physicianis fees.
  • UTP or U2P4 as a sputum induction agent will lower health care costs by avoiding the need for bonchoscopy.
  • the method of the present invention could be incorporated into a large-scale routine screening program (i.e., every 6 months to 1 year) for the general population or for populations at risk of developing lung cancer, so that lung cancer is detected as early as possible.
  • a novel method of early detection of lung cancer comprises using uridine 5'- triphosphate (UTP), P 1 , P 4 -(uridine 5'-) tetraphosphate (U2P4) or related compounds to induce a sputum sample from the lower lungs of an individual, then analyzing this sputum for the presence of labeled reagents selectively bound to substances in the sputum sample whose enhanced presence correlates with the development of lung cancer.
  • U2P4 uridine 5'- triphosphate
  • U2P4 uridine 5'-(uridine 5'-) tetraphosphate
  • the method of the present invention is particularly useful for individuals who are at high risk for lung cancer, including cigarette smokers or people who have been chronically exposed to airborne carcinogens such as asbestos, coal dust, tin ores, pesticides, fungicides, etc., or groups of people subject to occupational or environmental exposures such as steel workers, blast furnace operators, crop dusters, etc.
  • the method of the present invention could also be incorporated into a routine, periodic screening program (i.e., every 6-12 months) for the general population or high risk population.
  • the method involves: first, administrating a pharmaceutically effective amount of aerosolized UTP or U2P4 to the lungs of a subject at high risk for lung cancer to induce expectoration of sputum suitable for cytologic analysis; and second, analyzing this sputum for the presence of labeled reagents selectively bound to substances in the sputum sample whose presence correlates with the development of lung cancer.
  • a preferred method of sputum analysis entails Mab immunostaining to determine the enhanced presence of cell surface antigens which have been found to correlate with the development of lung cancer.
  • An additionally effective method of sputum analysis entails PCR analysis to detect gene mutations which correlate with the subsequent development of lung cancer.
  • Sputum is induced by administering to the patient a compound of Formula I, II, III or IV, or a pharmaceutically acceptable salt thereof, in an amount effective to allow the patient to expectorate or produce an amount of sputum from the lower lung sufficient for subsequent cytologic analysis:
  • Xi, X2, and X3 are each independently either O" or S-.
  • X2 and X3 are O".
  • Ri is O, imido, methylene, or dihalomethylene (e.g., dichloromethylene, diflouromethylene).
  • Ri is oxygen or difluoromethylene.
  • R2 is H or Br.
  • R2 is H.
  • Particularly preferred compounds of Formula I are uridine 5'-triphosphate [UTP] and uridine 5'-0-(3- thiotriphosphate) [UTP ⁇ S].
  • Formula II i.e., P 1 , P 4 -di(uridine-5') tetraphosphate [U2P4] is also a preferred embodiment of the invention.
  • Another compound of Formula II is P 1 , P 4 -di(adenosine-5') tetraphosphate [A2P4].
  • the method of the present invention can also include administering a compound of Formula III (adenosine 5 1 triphosphate [ATP] or l,N 6 -ethenoadenosine 5'- triphosphate or adenosine 1-oxine 5'-triphosphate), or Formula IV (cytidine 5'- triphosphate [CTP] or 3,N 4 -ethenocytidine 5'-triphosphate).
  • a compound of Formula III adenosine 5 1 triphosphate [ATP] or l,N 6 -ethenoadenosine 5'- triphosphate or adenosine 1-oxine 5'-triphosphate
  • Formula IV cytidine 5'- triphosphate [CTP] or 3,N 4 -ethenocytidine 5'-triphosphate
  • B is uracil or adenine or l,N 6 -ethenoadenine, attached as in Formulae I and III.
  • Ri, Xi, X 2 , and X3 are defined as in Formula I.
  • R3 and R4 are H while R2 is nothing and there is a double bond between N- 1 and C-6 (adenine), or
  • R3, .and R4 are H while R 2 is O and there is a double bond between N-l and C-6 (adeninel -oxide), or
  • Ri, Xi, X2, and X3 are defined as in Formula I.
  • R5 and R 6 are H while R7 is nothing and there is a double bond between N- 3 and C-4 (cytosine), or,
  • an optional first step is to cytologically analyze the sputum to confirm that the sputum originated in the deep lung. This is typically done by looking for the presence of alveolar macrophages which localize in the deep lung and are found in sputum originating from the deep lung. Labeled reagents are then applied to the sputum to determine the presence of tumor-associated substances.
  • One such labeled reagent is a monoclonal antibody (Mab) used to detect cell surface proteins associated with the development of lung cancer.
  • Another such labeled reagent is an oligonucleotide probe which selectively hybridizes with mutated gene sequences amplified through PCR, such mutated gene sequences correlating with the development of lung cancer.
  • the method of the present invention may be used to detect lung cancer at an early stage, while it is typically still surgically rescectable and potentially curable.
  • the method comprises using nebulized uridine 5'-triphosphate (UTP) (or related compounds) which is inhaled into the airways by a subject at risk for lung cancer in order to induce expectoration of sputum.
  • UDP nebulized uridine 5'-triphosphate
  • the method of the present invention further comprises analyzing exfoliated lung epithelial cells contained in the sputum specimen by means of immunocytochemical staining with monoclonal antibodies (Mabs) reactive to specific cancer-associated antigens.
  • Mabs monoclonal antibodies
  • UTP is well suited as a sputum induction agent because of its ability to increase mucociliary clearance (MCC).
  • MCC mucociliary clearance
  • UTP increases MCC in three ways: (1) by increasing the ciliary beat frequency of cilia on the surface of luminal epithelia cells, (2) by increasing the secretions of mucirts by goblet cells, .and (3) by increasing the chloride ion secretion and simultaneously increasing the secretion of water into the periciliary liquid layer by luminal epithelial cells, which would tend to lower the viscosity of the mucus.
  • UTP can still increase clearance of mucus in these patients by increasing mucin production and hydration of the mucus, therefore making the sputum easier to expectorate.
  • the compounds of the present invention are illustrated in Formulae I - IV.
  • uridine 5'-triphosphate UTP
  • uridine 5'-0-(3-thiotriphosphate) UTP ⁇ S
  • 5-bromo-uridine 5'-triphosphate 5-BrUTP
  • UTP may be made in the manner described in Kenner, et al., /. Chem. Soc.
  • Compounds illustrative of the compounds of Formula II include P 1 ,? 4 - di(uridine-5') tetraphosphate (U2P4) or P 1 , P 4 -di(adenosine-5') tetraphosphate (A2P4).
  • U2P4 can be prepared by methods similar to that described in C. Vallejo, et al., Biochem. Biophys. Ada 438, 304-09 (1976).
  • Compounds illustrative of the compounds of Formula III above include (a) adenosine 5'-triphosphate (ATP) and (b) l,N 6 -ethenoadenosine 5'-triphosphate.
  • Compounds illustrative of the compounds of Formula IV above include (a) cytidine 5 '-triphosphate and (b) 3,N 4 -ethenocytidine 5 '-triphosphate. These compounds can be made in accordance with known procedures, or variations thereof which will be apparent to those skilled in the art. For example, phosphorylation of nucleosides by standard methods such as D. Hoard and D. Ott, /. Am. Chem. Soc. 87, 1785-1788 (1965); M.
  • UTP, ATP, CTP, A 2 P 4 , 3,N 4 -ethenocytidine triphosphate, 1,N 6 - ethenoadenine 5'-triphosphate, adenosine 1-oxide 5'-triphosphate, ATP ⁇ S, ATP ⁇ S, ATP ⁇ S, AMPPCH2P, AMPPNHP, N 4 - ethenocytidine and W- ethenoadenosine are commercially available, for example, from Sigma Chemical Company, PO Box 14508, St. Louis, MO 63178.
  • the active compounds of Formulae I - IV may be administered by themselves or in the form of their pharmaceutically acceptable salts, e.g., an alkali metal salt such as sodium or potassium, an alkaline earth salt, or an ammonium and tetraalkyl ammonium salts, NX4+ (wherein X is O-4 alkyl).
  • pharmaceutically acceptable salts are salts that retain the desired biological activity of the parent compound and do not impart undesired toxicological effects.
  • the sputum induction agent of the present invention may be administered to the airways by a variety of suitable means, but is preferably administered by means of nebulizing a liquid/ liquid suspension via any of several commercially available nebulizer (e.g., jet, ultrasound, etc.).
  • nebulizer e.g., jet, ultrasound, etc.
  • One such nebulizer is the Pari LC Jet PlusTM .
  • the dosage of active compound to hydrate mucous secretions and stimulate ciliary beat frequency in the airways will vary depending on the state of the subject, but generally an effective amount is the amount sufficient to achieve concentrations of active compound on the airway passages of the subject of from about 10 - to about 10 moles/liter (e.g., for UTP, from about 0.00005 mg/mL to about 50 mg/mL), and more preferably from about 10 - 6 to about 10 ⁇ moles/liter (e.g., for UTP, from about 0.0005 mg/mL to about 50 mg/mL).
  • the sputum may be homogenized, concentrated, and placed in Saccomannois preservative solution (SPS, 2% polyethylene glycol in 50% ethanol) using standard methods as described in Saccomanno, et al., Ada Cytol. 2, 305-10 (1963).
  • SPS Saccomannois preservative solution
  • An optional next step is to cytologically analyze the sputum to confirm that the sputum originated in the deep lung. This is typically done by looking for the presence of alveolar macrophages which usually localize in the deep lung and which are usually found in sputum originating in the deep lung.
  • immunocytochemical staining is performed on aliquots of the preserved samples to determine the presence of tumor-associated biomarker antigens on the surface of exfoliated lung epithelial cells contained in the sputum.
  • a description of the immunostaining procedure, the two monoclonal antibodies used, and the computer-assisted cytometry employed is contained in M. Tockman, et al., Diagnostic Cytopathology 9, 615-622 (1993).
  • sputum samples may be incubated with murine nomonclonal antibodies 624H12 (American Type Culture Collection Accession Number HB10479, Rockville, MD) and 703D4 (ATCC Ace. No.
  • Mab 703DR a murine I G2b monoclonal antibody, targets an antigen associated with small cell lung cancer; this antigen is a 31 kD protein which is homologous to the ot2 ⁇ splice variant of the RNA-binding domain of the pre- mRNA-binding protein, called hnRNP.
  • the immunostained tumor-associated antigens may be detected by any suitable screening technique, such as immunoassay, immunoprecipitation assay, or immunohistochemistry assays.
  • a preferred embodiment entails image analysis of the immunostained sputum samples by means of a computer-assisted imaging program. The intensity of light transmitted through the immunostained sputum specimens is evaluated by a computer program at two frequencies of light. Positive and negative controls .are stained and anaylzed with each run. As a result of immunostaining, the cytoplasm of each cell acquires a brown (diaminobenzidine, i.e., DAB) color, while the nucleus has received a blue
  • DAB diaminobenzidine
  • the DAB stain has a maximum transmission at 600 nanometers (nm), and a transmission minimum at 570 nm. At 600 nm, the cytoplasm of a positively staining cell appears relatively translucent, while at 510 nm the cytoplasm appears opaque. Each image should be calibrated and corrected for shading.
  • An alternative method of sputum analysis involves PCR amplification and detection of mutated gene sequences, the presence of which correlates with the development of lung cancer.
  • PCR analysis is described in L. Mao, et al., Cancer Res. 54, 1634-37 (1994).
  • Sputum DNA is amplified by PCR with primers for K-ras and p53 that contain EcoRI sites to facilitate cloning.
  • products are cleaved with EcoRI and ligated to Lambda Zap II (Stratagene, La Jolla, CA).
  • XLI-blue cells infected with bacteriophage are plated on L-Agar at a density of 500-3000 plaques/ plate, transferred to nylon membranes, and hybridized with oligonucleotides specific for wild type or mutant K-ras and p53.
  • the oligonucleotides used for hybridizations are labeled with [ ⁇ - 32 P] ATP and hybridized according to the method of D. Sidransky, et al., Science 252, 706-709 (1991). Oligonucleotides used for detection include:
  • WT rus 5'-GGAGCTGGTGGCGTAGGCAA-3'
  • Val i2 mutant 5'-GGAGCTGTTGGCGTAGGCAA-3'
  • WT p53 5'-ATGGGCGCCATGAACCGG-3' His 273 mutant: 5'- i GAGGTGCATGTTTGTG-3'
  • Gly 281 mutant 5'-TGGGAGAGGCCGGCGCA-3'
  • TMV trachael mucus velocity
  • UTP and U2P4 produced significant dose-related effects on tracheal mucus velocity.
  • the doses ranged from 4 to 400 ⁇ mole. Both compounds had their maximal effects at a dose of 400 ⁇ mole (4 ml of lO ⁇ M).
  • This single-center, Phase I Unit study was a randomized, double-blind, evaluation of escalating, single doses of aerosolized UTP in 48 healthy male volunteers.
  • Four successive groups of 12 volunteers were enrolled at each dose level and were randomized in a 2:1 fashion to receive UTP or placebo.
  • Placebo was normal saline and the four dose levels of UTP evaluated in this study were: 0.5 mg/mL, 5 mg/mL, 25 mg/mL and 45 mg/mL.
  • 4 mL of placebo or 4mL of the appropriate UTP solutions were placed in a nebulizer for aerosolization.
  • Each subject was randomly assigned to receive a single dose of either UTP or placebo.
  • Each dose consisted of 4mL of placebo or the appropriate solution of UTP (0.5, 5, 25 or 45 mg/mL) and was administered using a jet nebulizer (Pari LC PLUSTM) powered by a portable compressor set at a flow rate of 14 L/min. Inhalation of placebo or UTP took approximately 8-15 minutes.
  • - Efficacy Results Sputum was collected for the purpose of cytological examination to determine whether the sample contained alveolar macrophages. The presence of alveolar macrophages in a sputum sample indicates that the sample is a quality specimen arising from deep within the lungs (not simply salivary secretions).
  • sputum was collected for cytological examination at the following times: baseline (pre-dose), immediately post-dosing, post-dosing to 4 hours, 4 hours to 8 hours post-dosing, 8 hours to 12 hours post-dosing and upon rising the following day.
  • FIG. 5 illustrates the timecouse for the effect of placebo and UTP on the percentage of sputum samples determined to be positive for alveolar macrophages (AM). Although the majority of the subjects (pre-dose period) could produce a sputum sample at baseline (44/48 subjects spontaneously and 47/48 subjects produced a sample following a simple deep breath and cough maneuver); only -30% of these samples were considered to be a quality sputum sample as evidenced by cytological examination (presence of AMs).
  • placebo normal saline
  • the effect of UTP to improve the percentage of samples positive for alveolar macrophages was also evident at the second time point (end of dosing to 4 hours post-dosing); 37% of samples were positive following administration of placebo versus 57% following UTP.
  • the doses of placebo and UTP were administered as 4 mL of the appropriate solution placed into a nebulizer for aerosolization.
  • the purpose of this study was to determine if UTP could enhance the ability of smokers to expectorate sputum over placebo.
  • Chronic smokers are at significant risk to develop lung cancer.
  • one agent frequently used for inducing sputum induction is saline; therefore, the placebo chosen for this study was normal saline.
  • Each subject was randomly assigned to receive a single dose of either UTP (one of four doses) or placebo.
  • Each dose consisted of 4mL of the appropriate solution (0.5, 25 or 45 mg/mL) and was administered using a jet nebulizer (Pari LC PLUSTM) powered by a portable compressor set at a flow rate of 14 L/min. Inhalation of placebo or UTP took approximately 8-15 minutes.
  • the amount of sputum expectorated was collected at baseline (pre-dose), immediately post-dosing, post-dosing to 4 hours, 4 hours to 8 hours post-dosing, 8 hours to 12 hours post-dosing and upon arising the following day.
  • Each subject was randomly assigned to receive multiple daily doses (three times a day for three consecutive days) of placebo and UTP with at least 1 week between the two dosing periods.
  • Each dose consisted of 4mL of the 45 mg/mL solution or placebo (normal saline) administered using a jet nebulizer (Pari LC PLUSTM) powered by a portable compressor set at 14 L/min. Inhalation of placebo or UTP took approximately 8-15 minutes.
  • inhalation of placebo did not increase the amount of sputum expectorated immediately post-dosing during any of the three days of dosing (comparison of sputum weights at pre-dosing to immediately post-dosing).
  • inhalation of UTP consistently increased the amount of sputum expectorated immediately post-dosing on all three days of dosing (comparison of sputum weights at pre-dosing to immediately post-dosing); the magnitude of the increase in the amount of sputum expectorated (pre to post dosing of UTP) was consistent on each of the three days for the UTP dosing period.
  • FIG 8 illustrates the cytology data for the sputum expectorated by smokers receiving UTP versus placebo.
  • smokers receiving UTP were more likely to produce a sputum sample containing alveolar macrophages (AM) on any of the 3 days than smokers receiving placebo (the difference between placebo and UTP was most pronounced on Day 2).
  • placebo the difference between placebo and UTP was most pronounced on Day 2.
  • This single center study conducted at a major academic center in the US, was a randomized, double-blind, cross-over evaluation of escalating, single inhaled doses of UTP in patients with chronic bronchitis.
  • Patients enrolled in this study had to meet the American Thoracic Society definition of chronic bronchitis (excessive mucous production over 3 months of the year, for at least 2 successive years). Patients were included that had mild to moderate airflow obstruction (forced expiratory volume over 1 second >65% of predicted at study entry). A total of 26 (14 females and 12 males) patients were enrolled in this study and the majority were currently smoking.
  • each subject was randomly assigned to receive single inhaled dose of placebo (normal saline) or one of five doses of UTP on two separate days.
  • each dose consisted of 4mL of 2.5, 5, 15, 25, and 45 mg/mL solution or placebo (normal saline) administered using a jet nebulizer (Pari LC PLUS) powered by a portable compressor set at 14 L/min. Inhalation of placebo or UTP took approximately 8-15 minutes.
  • Figure 9 illustrates the effect of placebo versus UTP on the amount of sputum expectorated (weight in grams) at two time points: baseline (spontaneous expectoration) versus immediately to 5 minute post-dosing.
  • UTP all doses combined
  • the ability of UTP to enhance the amount of sputum expectorated over placebo and baseline was also quite evident at the later timepoint of 6 minute - 30 minute post-dosing, as shown in Figure 10.
  • Figure 11 shows that at the 31 minute to discharge time point there was essentially no difference between the effect of UTP and placebo on the amount of sputum expectorated indicating that the effect of UTP is manifest over a short timeframe, consistent with the previous studies.
  • UTP represents a safe, rapid and non-invasive approach to obtaining a quality sputum sample for the purposes of screening for abnormal cells.

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Abstract

Procédé servant à détecter le cancer des poumons de façon précoce. Ce procédé consiste à utilise uridine 5'-triphosphate (UTP) ou des composés apparentés provoquant l'expectoration dans le but d'obtenir un spécimen de crachat provenant des poumons d'un individu, puis à utiliser des réactifs marqués, afin de détecter des substances dans les crachats, dont la présence est associée à l'évolution d'un cancer des poumons. Des procédés de détection de substances associées au cancer dans les crachats comprennent l'immuno-coloration par des anticorps monoclonaux afin de détecter des antigènes cellulaires associés au cancer, ou l'analyse par PCR afin d'amplifier et de détecter des mutations de gènes associés au cancer. On administre l'agent d'UTP à un individu au moyen d'une solution liquide, d'une suspension liquide sous forme nébulisée ou au moyen d'un agent d'inhalation sous forme de poudre sèche.
PCT/US1997/018137 1996-10-08 1997-10-08 Procede de detection precoce du cancer des poumons par provocation de l'expectoration et analyse des crachats afin de detecter des substances associees au cancer WO1998015835A1 (fr)

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Cited By (9)

* Cited by examiner, † Cited by third party
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US6319908B1 (en) 1996-07-03 2001-11-20 Inspire Pharmaceuticals, Inc. Method for large-scale production of di(uridine 5′-tetraphosphate) and salts thereof
US6703376B2 (en) 1996-07-23 2004-03-09 Inspire Pharmaceuticals, Inc. Use of uridine triphosphates and related compounds for the prevention and treatment of pneumonia in immobilized patients
US6713458B1 (en) 1997-07-25 2004-03-30 Inspire Pharmaceuticals, Inc. Therapeutic uses of di(uridine 5′)-tetraphosphate and salts thereof
US6872710B2 (en) 1997-07-25 2005-03-29 Inspire Pharmaceuticals, Inc. Di(uridine 5′)-tetraphosphate and salts thereof
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US6319908B1 (en) 1996-07-03 2001-11-20 Inspire Pharmaceuticals, Inc. Method for large-scale production of di(uridine 5′-tetraphosphate) and salts thereof
US6703376B2 (en) 1996-07-23 2004-03-09 Inspire Pharmaceuticals, Inc. Use of uridine triphosphates and related compounds for the prevention and treatment of pneumonia in immobilized patients
US7091334B2 (en) 1997-07-25 2006-08-15 Inspire Pharmaceuticals, Inc. Method for large-scale production of di(uridine 5′)-tetraphosphate and salts thereof
US6765090B2 (en) 1997-07-25 2004-07-20 Inspire Pharmaceuticals, Inc. Method for large-scale production of di(uridine 5')-tetraphosphate and salts thereof
US6872710B2 (en) 1997-07-25 2005-03-29 Inspire Pharmaceuticals, Inc. Di(uridine 5′)-tetraphosphate and salts thereof
US6713458B1 (en) 1997-07-25 2004-03-30 Inspire Pharmaceuticals, Inc. Therapeutic uses of di(uridine 5′)-tetraphosphate and salts thereof
US7132410B2 (en) 1997-07-25 2006-11-07 Inspire Pharmaceuticals, Inc. Di(uridine 5′-)tetraphosphate and salts thereof
US7432252B1 (en) 1997-07-25 2008-10-07 Inspire Pharmaceuticals, Inc. Method of promoting cervical and vaginal secretions
US7939510B2 (en) 1997-07-25 2011-05-10 Inspire Pharmaceuticals, Inc. Di(uridine 5′-)tetraphosphate and salts thereof
US7018985B1 (en) 2000-08-21 2006-03-28 Inspire Pharmaceuticals, Inc. Composition and method for inhibiting platelet aggregation
US7618949B2 (en) 2000-08-21 2009-11-17 Inspire Pharmaceuticals, Inc. Drug-eluting stents coated with P2Y12 receptor antagonist compound
US7109181B2 (en) 2001-06-25 2006-09-19 Inspire Pharmaceuticals, Inc. Joint lubrication with P2Y purinergic receptor agonists
US7256183B2 (en) 2001-11-06 2007-08-14 Inspire Pharmaceuticals, Inc. Method for treating or preventing inflammatory diseases

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