Patent Application
20200018753
This invention relates generally to methods for diagnosis of increased risk for preterm delivery during pregnancy. The invention also relates to compositions and methods for applications of specific prostaglandin D2 receptor (DP1 and DP2) antagonists as novel tocolytic therapeutic agents. The invention relates to applications wherein women with a diagnosis of preterm labor or at significant risk for preterm delivery, based on medical history and/or other factors, might be tested. The invention also relates to applications wherein pregnant women generally may be tested at 28 weeks of pregnancy to assess the likelihood of preterm delivery.
Preterm birth is a leading cause of neonatal morbidity and mortality, accounting for 35% of infant healthcare spending and 10% of all childcare spending in the United States. In addition, medical costs from birth to one year of age are 10-fold greater for babies born premature and/or at low-birth weight when compared to those for babies born full-term. The rate of preterm birth has steadily increased by 20% since 1990, with 12.5% of all U.S. births being preterm births. Thus, there is a clear and significant need to understand the biological and molecular mechanisms leading to preterm birth, as well as to improve a physician's ability to predict which women are at risk for preterm delivery.
There are approximately 500,000 preterm in births per year in the United States, of which approximately 30-70% are associated with an underlying infectious process. Infection is a major risk factor for pre-term birth after the most consistently identified risks factors, which include: a history of preterm birth, current multi-fetal pregnancy, and some uterine and/or cervical abnormalities. The ability to identify high-risk pregnancies at an early stage will lead to a significant reduction in the incidence of preterm birth and most likely reduce the incidence of fetal demise, decrease the rate of cerebral palsy, and other neuro-developmental delays. By improving our ability to identify those at risk of preterm birth, we also provide for the development of new and more effective treatments for preterm labor.
There is little overlap between the current biomarkers of preterm birth, suggesting that these various biomarkers predict different pathways that lead to preterm birth, and suggesting that the creation of a highly predictive biomarker would be beneficial. Currently, there are a handful of biomarkers which attempt to predict preterm labor in advance in order to prevent preterm delivery. These biomarkers include fetal fibronectin, salivary estriol, decidual proteins, and endocrinelparacrine. Fetal fibronectin and salivary estriol have been examined in some detail, with fetal fibronectin now being made available commercially. The fetal fibronectin test is based on the detection of fetal fibronectin, a fetal-specific glycosylated form of fibronectin. The theory is that increased amounts of fetal fibronectin signal the disturbance of the junction of the fetal membranes and the placental deciduas and therefore predicts delivery. In symptomatic pregnant women, the test has only limited effectiveness (58% sensitivity) to predict preterm delivery before the completion of 37 weeks gestation. The main benefit of the present test is the ability to exclude the possibility of preterm delivery (85% specificity) and to avoid unnecessary interventions. Parturition is comprised of 5 events: (1) membrane rupture; (2) cervical dilation; (3) myometrial contaction; (4) placental separation; and (5) uterine involution. While previously available tests might focus on detection of only a subset of these five events (and thus potentially lead to a heightened likelihood for “false positives” in the detection of a purported increased likelihood of preterm birth conditions), the present invention relates to a novel method for heightened sensitivity (e.g., detection of a unified factor relating to all 5 significant steps in parturition), coupled with a more practical effectiveness to diagnosis and possibly treat or prevent a broader range of potential preterm birth conditions. See, U.S. Pat. Nos. 4,919,889; 5,096,830; 5,223,440; 5,281,522; 5,431,171; 5,468,619; 5,516,702; 5,597,700; 5,641,636; 5,650,394; 5,698,404; 5,783,396; 5,877,029; 5,891,722; 5,968,758; 6,126,597; 6,126,616; 6,140,099; 6,149,590; 6,267,722; 6,312,393; 6,394,952; 6,544,193; 6,556,977; 6,610,480; 6,678,669; 6,867,051; 6,875,567; 6,936,476; 7,041,063; 7,191,068; 7,228,295; 7,270,970; 7,403,805; 7,488,585; 7,524,636; 7,528,133; 7,654,957; 7,709,272; 7,756,559; 7,809,417; 7,811,279; 7,863,007; 7,943,294; 8,060,195; 8,068,990; 8,114,610; 8,133,859; 8,160,692; 8,366,640; 8,372,581; 8,501,688; 8,517,960; 8,552,364; U.S. Pat. Pub. Nos. 20010023419; 20010025140; 20010053876; 20020031513; 20020046054; 20020049389; 20030004906; 20030099651; 20030105731; 20030113319; 20030139687; 20040014063; 20040039297; 20040039298; 20040078219; 20040100376; 20040197930; 20040241752; 20040266025; 20050101841; 20050131287; 20050163771; 20050260683; 20050277912; 20060008923; 20060014302; 20060024722; 20060024723; 20060024724; 20060024725; 20060024757; 20060040337; 20060166242; 20060166295; 20060204532; 20060240495; 20060240498; 20070016074; 20070128589; 20070142718; 20070161125; 20080009552; 20080090759; 20080254479; 20080299594; 20090036761; 20090055099; 20090058072; 20090068692; 20090081206; 20090171234; 20090227036; 20090299212; 20100008911; 20100017143; 20100029006; 20100093621; 20100145180; 20100311190; 20100318025; 20110028807; 20110040161; 20110065139; 20110090048; 20110159533; 20110166070; 20110184254; 20110237972; 20110312927; 20110312928; 20120009609; 20120040356; 20120046261; 20120052595; 20120082598; 20120157422; 20120196285; 20120202740; 20120238469; 20120238894; 20120270747; 20130029957; 20130053670; 20130071865 20130171672; 20130177485; and 20130225922, each of which is expressly incorporated herein by reference.
Measurement of changes in Lipocalin-type prostaglandin D2 synthase (L-PGDS) levels in human cervicovaginal fluid has been suggested as a possible indicator of membrane rupture during pregnancy. Shiki Y, Shimoya K, Tokugawa Y, Kimura T, Koyama M, Azuma C, Murata Y, Eguchi N, Oda H, Urade Y 2004 Changes of lipocalin-type prostaglandin D synthase level during pregnancy. J Obstet Gynaecol Res 30:65-70 (37), each of which is expressly incorporated herein by reference.
Rachel J. A. Helliwell, Jeffrey A. Keelan, Keith W. Marvin, Linda Adams, Maxwell C. Chang, Ashmit Anand, Timothy A. Sato, Simon O'Carroll, Tinnakom Chaiworapongsa, Roberto J. Romero, and Murray D. Mitchell, “Gestational Age-Dependent Up-Regulation of Prostaglandin D Synthase (PGDS) and Production of PGDS-Derived Antiinflammatory Prostaglandins in Human Placenta”, The Journal of Clinical Endocrinology & Metabolism, vol. 91 no. 2 pp. 597-606 (Feb. 1, 2006), each of which is expressly incorporated by reference, discuss that The two isoforms of PGDS (L-PGDS and H-PGDS) immunolocalized to distinct regions of human term and preteen gestational tissues. In placental villous tissues, specific labeling for L-PGDS was identified in the syncytiotrophoblast layer of both preterm and term placenta, with prominent staining of the apical membrane. At term, cytoplasmic syncytial staining was readily apparent in the syncytium, giving rise to a characteristic beaded appearance. Similarly, strong H-PGDS immunolabeling was observed in the syncytial layer of the preterit” placenta. However, cellular localization of H-PGDS appeared to change with gestational age, and by term, immunoreactive H-PGDS was mainly localized to cells lining the villous capillaries, with little or no labeling observed in the syncytium. Labeling was completely absent in the corresponding negative controls in which primary antibody was preincubated with a 10-fold excess of blocking peptide or was omitted completely. In gestational membranes collected from both preterm and term pregnancies, strong immunolabeling for L-PGDS was observed in the cells of all tissues, including amnion epithelial, reticular, chorionic trophoblast, and dccidual cells. In contrast, only very weak labeling of the gestational membranes was observed for H-PGDS. No labeling was observed in the corresponding negative controls. No significant staining of infiltrating leukocytes was apparent, although on some slides occasional cells positive for L- or H-PGDS were visualized among maternal blood cells. There was no statistical difference in the net amount of H-PGDS in any tissue as a result of the onset of labor at term or the presence of intrauterine infection preterm. Immunoreactive L-PGDS was undetectable in gestational tissues. L-PGDS mRNA was detectable in amnion, choriodecidual, and villous placental samples There was no significant effect of preterm intrauterine infection or term labor on the level of expression of L-PGDS mRNA. L-PGDS mRNA levels were significantly higher in the choriodecidua and placenta than in the amnion. There was no significant difference in the level of expression between the choriodecidua and placenta. The expression of H-PGDS mRNA was detectable in all three tissues, with no significant effect of preterm intrauterine infection or labor at term. Analyzing the combined data for each tissue revealed that the relative expression of H-PGDS mRNA was lowest in the amnion and choriodecidua, with a significantly higher level of expression in the villous placenta.
See also, Olson D M, Ammann C, “Role of the prostaglandins in labour and prostaglandin receptor inhibitors in the prevention of preterm labour”, Front Biosci.; 12:1329-43 (Jan. 1, 2007)., and Pirianov G, Waddington S N, Lindstrom T M, Terzidou V, Mehmet H, Bennett P R, “The cyclopentenone 15-deoxy-delta12,14-prostaglandin J2 delays lipopolysaccharide-induced preterm delivery and reduces mortality in the newborn mouse”, Endocrinology 150(2):699-706 (Epub 2008 October 9, February 20909), each of which is expressly incorporated herein by reference.
U.S. Pat. No. 7,399,596 and US 20070020609, expressly incorporated herein by reference, discusses using L-PGDS levels to predict pregnancy-induced hypertension. See also, U.S. Pat. Nos. 7,902,373, 7,794,953, 7,582,643, 7,314,727, 7,109,044, 6,790,635, 6,605,705, 6,410,583, 20110021599, 20100323911, 20100251394, 20080233597, 20080227113, 20070196864, 20070003992, 20040038314, 20030190678, each of which is expressly incorporated herein by reference.
Specific DP1/DP2 agonists and antagonists are available, e.g., BW245C (5-(6-carboxyhexyl)-1-(3-cyclohexy1-3-hydroxypropyl-hydantoin), AS702224, TS-022, 15R-methyl-PGD2, 13-14-dihydro-15-keto-PGD2, AM156, AM206, L-745870, 15R-PGD(2), MK-0524, BWA868C, BW24-SC, BAY-u3405, 15-deoxy-Delta12,14-prostaglandin J2 (15d-PGJ2), 11-deoxy-11-methylene PGD2, G06983 (PKCα,Δ,ϵ,ζ, G06976 (PKCα), GF10923X (PKCα,Δ,ϵ), LY333531 (PKC β), SB203580 (p38MAPK), SB203580, CD200, FGF18, GPRCSD, GPR49, LRRC15, Serpin A, CDT6, BMP2, LHX2, THBS1, MYCN, NR4A2, MEST, TM4SF1, CRLF1, TNFRSF12A, SELENBP1, GPR161, HEPH, FZD7, and CLIC4, CCL18, Col11A1, Col11Al, CD4, Cdla, FCER1A, HLA-C, HLA-DPA1, IGF I, GPR105, PDGFRL, ADRA2A, CCL19, CORN, 16-phenoxy-17,18,19,20-tetranor PGD2 N-cyclopropylamide, 16-phenoxy-1718,19,20-tctranor PGD1 N-cyclopropylmethylamide, 16-phenoxy-17,18,19,20-PGD1N-(1,3-dihydroxypropan-2-yl))amide; 17-phenyl-18,19,20-trinor PGD2 N-cyclopropylamide, 17-phenyl-18,19,20-trinor PGD1N-cyclopropylmethylamide, 17-phenyl-18,19,20-trinor PGD2N-(1,3-dihydroxypropan-2-yl))amide; 16-(3-chloropheny1)-17,18,19,20-tetranor PGD2 N-cyclopropylamide, 16-(3-chloropheny0-17,18,19,20-tetranor PGD1N-cyclopropylmethylamide, 6-(3-chloropheny1)-17,18,19,20-tetranor PGD1 N-(1,3-dihydroxypropan-2-yl))amide, (Z)-isopropyl 7-((R)-2-((R)-3-hydroxy-5-phenylpentyl)-5-oxocyclopent-2-enyphept-5-enoate, (Z)-isopropyl 7-((R)-2-((R,E)-3-hydroxy-4-(3-(trifluoromethyl)phenoxy) but-l-eny1)-5-oxo-cyclopent-2-enyl)hept-5-enoate, (Z)-N-ethyl-7-((R)-2-((R,E)-3-hydroxy-4-(3-(trifluoromethyl)phenoxy)but-l-eny1)-5-oxocyclopent-2-enyl)hept-5-enamide, (Z)-N-ethyl-7-((R)-2-((S,E)-3-hydroxy-5-phenylpent-1-eny1)-5-oxocyclopen-t-2-enyl) hept-5-enamide, (Z)-7-((R)-2-((R,E)-3-hydroxy-4-(3-(tritluoromethyl)phenoxy)but-l-eny1)-5-oxocyclopent-2-enyl)hept-5-enoic acid, (Z)-7-((R)-2-((R,E)-3-hydroxy-4-(3-(trifluoromethyl) phenoxy)but-l-eny1)-5-oxocyclopent-2-enyl)-N-methylhept-5-enamide, (Z)-7-((R)-2-((R,E)-4-(3-chlorophenoxy)-3-hydroxybut-1-eny1)-5-oxocyclope-nt-2-enyl)hept-5-enoic acid, (Z)-isopropyl 7-((R)-2-((R,E)-4-(3-chlorophenoxy)-3-hydroxybut-l-eny1)-5-oxocyclopent-2-enyl) hept-5-enoate, (Z)-7-((R)-2-((R,E)-4-(3-chlorophenoxy)-3-hydroxybut-l-eny1)-5-oxocyclopent-2-enyl) -N-methylhept-5-enamide or a pharmaceutically acceptable salt, hydrate, solvate, prodrug or metabolite thereof. These agents may be used alone or in combination, and may be administered concurrently or sequentially. See also, 2011/0144160, 2011/0130453, 2011/0112134, 2011/0098352, 2011/0098302, 2011/0071175, 2011/0060026, 2011/0034558, 2011/0028717, 2011/0021599, 2011/0021573, 2011/0002866, 2010/0330077, each of which is expressly incorporated herein by reference.
This invention describes, for example, methods to apply and correlate the clinical measurement of L-PGDS concentrations of cervical vaginal secretions from pregnant women with the attendant likelihood of term or preteen delivery.
The invention also describes methods and compositions, specifically prostaglandin D2 (DP1 and DP2) receptor antagonists or inhibitors, as potential tocolytic agents.
It is therefore an object to provide a method for assessing the likelihood of preterm delivery for a pregnant woman, the method comprising collecting a cervical vaginal secretion sample from a pregnant woman; and measuring lipocalin-type prostaglandin D2 synthase (L-PGDS) concentration levels from the sample, wherein L-PGDS concentration levels correlate to and indicate an above or below average likelihood relative to the relevant standard population for preterm delivery.
According to one embodiment, an L-PGDS concentration level higher than 1.8 μg/ml indicates an increased likelihood for preterm delivery.
The method may advantageously be applied to a pregnant woman has intact amniotic membranes.
The cervical vaginal secretion may be collected with a sponge.
Based on the L-PGDS level, at least one prostaglandin D2 receptor antagonist may be administered as a tocolytic agent. The at least one prostaglandin D2 receptor antagonist may comprises a DPI receptor antagonist and/or a DP2 receptor antagonist.
The L-PGDS concentration level may be determined using an antibody sandwich Enzyme-Linked ImmunoSorbent Assay (ELISA). The L-PGDS concentration levels may be normalized to protein levels in the cervical vaginal secretions for analysis.
Another object provides a method for treatment of preterm labor, comprising administration of a therapeutic amount of at least one prostaglandin DPI or prostaglandin DP2 receptor antagonist to a pregnant woman such that preterm delivery is avoided or mitigated.
The administration may be selectively in dependence on a measured L-PGDS level, for example in a cervical vaginal secretion.
The at least one prostaglandin DP1 or prostaglandin DP2 receptor antagonist comprises one or more agents selected from the group consisting of AM156 ({2′-[(cyclopropanecarbonyl-ethyl-amino)-methyl]-6-methoxy-4′-trifluoro-methyl-biphenyl-3-y11-acetic acid, sodium salt), and AM206 {2-Rbenzoyloxycarbonyl-ethyl-amino)-methy11-4-trifluoromethyl-phenyll-pyridin-3-yl)-acetic acid, sodium salt), MK-0524 ([(3R)-4-(4-Chloro-benzyl)-7-fluoro-5-(tnethylsulfony1)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-yThacetie Acid), AM-853 (2-(4-(4-(tert-butylcarbamoyl)-2-(2-chloro-4-cyclopropylphenyl sulfonamido)phenoxy)-5-chloro-2-fluorophenyl)acetic acid), BW868C (3-benzyl-5-(6-carbohexyl)-1-(2-cyclohexyl-2-hydroxyethylamino)-hydantoin), S-5751 ((Z)-7-[(1R,2R,3S,5S)-2-(5-hydroxy benzo[b]thiophen-3-ylearbonylamino)-10-norpinan-3-ylihept-5-enoic acid), and BAY-u3405 (Ramatroban, 3(R)-[[(4-fluorophenyl) sulphonyl]amino]-1 ,2,3,4-tetrahydro-9H-carbazole-9-propanoic acid).
Alternately, an L-PGDS inhibitor may be used, for example, AT-56 (4-dibenzo[a,d]cyclohepten-5-ylidene-1-[4-(2H-tetrazol-5-yl)-butyl]-piperidine).
Resveratrol has been shown to reduce formation of Prostaglandin D2 presumably by inhibiting cyclooxygenase (COX), especially COX-2. See, Lena Wendeburg, Antonio Carlos Pinheiro de Oliveira, Harsharan S Bhatia, Eduardo Candelario-Jalil and Bernd L Fiebich, “Resveratrol inhibits prostaglandin foiniation in IL-1β-stimulated SK-N-SH neuronal cells”, J. Neuroinflammation 2009, 6:26, expressly incorporated herein by reference. Therefore, resveratrol or another non-teratogenic COX-2 inhibitor may advantageously be employed.
The at least one prostaglandin DP1 or prostaglandin DP2 receptor antagonist is administered orally, by inhalation, through a mucous membrane, transdermally, topically, rectally, or parenterally.
The at least one prostaglandin DP1 or prostaglandin DP2 receptor antagonist may be administered to achieve concurrent therapeutic effect with another tocolytic agent which is not a prostaglandin D2 receptor antagonist. For example, the other tocolytic agent may comprise a β2 agonist, such as terbutaline, Ritodrine, Fenoterol, or Salbutamol. The other tocolytic agent may also comprise a non-steroidal anti-inflammatory drug cyclooxygenase inhibitor, such as Indomethicin or sulindac. The tocolytic may be a calcium channel blocker, such as nifedipine, a myosin light chain inhibitor such as magnesium sulfate, or an axytocin inhibitor such as Atosiban.
A further object provides a pharmaceutical formulation comprising a therapeutic amount of a non-teratogenic comprising at least one prostaglandin DP1 or prostaglandin DP2 receptor antagonist and/or an L-PGDS inhibitor, in an effective amount to delay preterm delivery in a pregnant human when administered in repeated doses. The at least one prostaglandin DP1 or prostaglandin DP2 receptor antagonist may comprise, for example, one or more agents selected from the group consisting of AM156 ({2′-[(cyclopropanecarbonyl-ethyl-amino)-methy1]-6-methoxy-4′-trifluoro-methyl-biphenyl-3-yll-acetic acid, sodium salt), and AM206 (5-{2-[(benzoyloxycarbonyl-ethyl-amino)-methyl]-4-trifluoromethyl-phenyll-pyridin-3-yl)-acetic acid, sodium salt), MK-0524 ([(3R)-4-(4-Chloro-benzyl)-7-fluoro-5-(methylsulfonyl)-1,2,3,4-tetrahydrocyclopenta[b]indol-3-y11-acetic Acid), AM-853 (2-(4-(4-(tert-butylcarbamoyl)-2-(2-chloro-4-cyclopropylphenyl sulfonamido)phenoxy)-5-chloro-2-fluorophenyl)acetic acid), BW868C (3-benzyl-5-(6-carbohexyl)-1-(2-cyclohexyl-2-hydroxyethylamino)-hydantoin), S-5751 ((Z)-7-[(1R,2R,3S,5S)-2-(5-hydroxy benzo[b]thiophen-3-yIcarbonylamino)-10-norpinan-3-yl]hept-5-enoic acid), and BAY-u3405 (Ramatroban, 3(R)-[[(4-fluorophenyl) sulphonyl]amino]-1,2,3,4-tetrahydro-9H-carbazole-9-propanoic acid), and the L-PGDS inhibitor may comprise AT-56 (4-dibenzo[a,d]eyelohepten-5-ylidene-144-(2H-tetrazol-5-yl)-butyl]-piperidine).
All publications, patents, and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference.
The novel features of the invention are set forth with particularity in the appended claims. A better understanding of the features and advantages of the present invention will be obtained by reference to the following detailed description that sets forth illustrative embodiments, in which the principles of the invention are utilized, and the accompanying drawings of which:
Cervical vaginal secretions (CVS) arc readily collected and constitute a′ minimally invasive procedure. During a sterile speculum examination of a pregnant woman, a Week-gel sponge is applied into the cervical os for approximately one minute. The sponge is then removed and placed in a buffer solution containing physiological saline, Trizma buffer, protease inhibitors (EDTA, phenylmethylsulfonyl fluoride, pepstatin) and antibacterial agents (such as sodium azide). This sample collection process is very similar to that used for other routine clinical assays (e.g., fetal fibronectin) that use cervical secretions for the starting sample material. Preferably, the buffer solution has a low protein concentration, e.g., is produced with a small amount or no bovine serum albumin (BSA). This BSA is not required to stabilize the L-PGDS, and may interfere with its measurement.
L-PGDS concentration levels from the cervical vaginal secretion samples are then determined by an antibody sandwich ELISA. As an initial step, antibody against L-PGDS raised in rabbits is purified by hydroxyapatite chromatography, followed by an immunoaffinity purification on an Affi-Gel 15 Gel column.
The purified antibody is then diluted to 10 μg/ml in 0.2M sodium carbonate buffer, pH 9.0, and 50 μl added to each well of an Nunc-Immuno plate with Maxi Sorp surface overnight at 4° C. The wells are then washed with 0.1% BSA/0.05% Tween 20 in PBS 4 times and blocked with 1% BSA in PBS for 1 hour. Diluted plasmas or L-PGDS standards (1.2 to 75 ng/ml) are incubated in Block for 90 minutes, then washed as above. A secondary anti-L-PGDS antibody, raised in chickens, at 4 ng/ml in wash buffer, is incubated in the wells for 1 hour and then washed, as above. Horseradish peroxidase-tagged goat anti-chicken antibody at 1:10,000 in wash buffer is added for 1 hour and then the plate is washed, as above, developed in a 0.01 3,3′, 5,5′ tetramethylbenzidine solution, stopped with 50 μl of IN sulfuric acid, and read at 450 nm in a microplate reader. L-PGDS concentration is determined by the standard curve obtained by plotting the absorbance versus the corresponding concentration of an L-PGDS standard.
This assay is also adapted to a BioPlex suspension array for similar quantitation. The assay is also amenable for development to run the samples on a 2-D PAGE and determine the particular L-PGDS isoforms present/absent from the CVS samples that may be indicative of an increased propensity toward preterm birth. It is understood and believed that an antibody to the specific isoform may be created to fine-tune the detection method.
In assessing test samples, L-PGDS concentrations determined by ELISA are normalized to protein levels in the CVS. Protein content of the samples is determined by BCA method, which has been commonly used to quantitate protein levels in cell lysates.
The new markers of the present invention are highly sensitive and specific (e.g., 90% or higher). In an preliminary study, at a level of 90% specificity, the sample size yields a 95% confidence level, indicating high precision, with a margin of error of about 5%. The collection of test data from pregnant women is ongoing and studies producing significant results have been established as demonstrated in the following Table 1.
Statistical analysis of this data indicates that high L-PGDS levels (i.e., L-PGDS levels above 1.8μ/ml) predict an increased likelihood of preterm delivery in human subjects. Graphical illustration of this analysis is depicted in
The pattern of elevated L-PGDS in preterm is generally consistent with the predicted rise. Because the L-PGDS level has a normal change over gestation, CVS samples are preferably taken and L-PGDS assays performed every two weeks.
A key advantage of this assay over the current fetal fibronectin (fFN) test is that the present assay would predict preterm birth over the entire gestational period up to parturition. A negative fFN, although very accurate, is only valid for a 2-week window. Interestingly, in the study reported below, there were several samples that were fFN negative, but ultimately went preterm. The present assay showed elevated L-PGDS, and was predictive for preterm birth.
The difference in the median values between the two groups is greater than would be expected by chance; there is a statistically significant difference (P=<0.001)
Animal studies conducted to determine the correlations between L-PGDS concentrations and birth viability and/or risk of preterm birth were conducted with L-PGDS knockout mice and transgenic L-PGDS overexpressor mice. Results are demonstrated in
Table 3 shows data demonstrating that in LPS-induced preterm birth, there is a three-fold increase in viable mouse pups/pregnancy in the L-PGDS knockouts and a 15-fold decrease in viable pups/pregnancy in the L-PGDS transgenic overexpressors when compared to the C57BL/6 controls. This implies that L-PGDS is casually related to preterm birth, and not merely correlated with it.
In a study of pregnant C57BL/6 mice implanted with Alzet osmotic pumps containing both DP1 and DP2 antagonists (BWA868C and 11-deoxy-11-methylene prostaglandin D2 at 1.0 μg/μl or PBS vehicle) were injected with lipopolysaccharide (LPS) (20 μg) at day 14 of pregnancy to induce preterm birth. In the LPS-only control group, 80% suffered adverse outcomes (premature birth or fetal death), and 20% had normal outcomes. In the experimental group, LPS plus DP1 and DP2 antagonist, 50% suffered adverse outcomes and 50% had normal outcomes. This demonstrates that administration of prostaglandin D2 receptor antagonists may be administered to pregnant mammals to interrupt preterm labor, and more particularly, to avoid preterm labor induced by infectious agents, especially those which produce lipopolysaccharides. Because the preterm labor pathway has common elements, it is likely that the prostaglandin D2 receptor antagonists will also act to block preterm labor from other causes.
Chemical compounds which influence the levels of L-PGDS in pregnant women might be administered as tocolytic therapeutic agents. Preferred embodiments of such effective tocolytic agents include prostaglandin D2 (DP1 or DP2) receptor antagonists, such as BWA868C, see, e.g., U.S. Pat. Nos. 6,395,499; 6,878,522; 6,884,593; 7,144,913; 7,217,725; 7,517,889; 7,642,249; 8,067,445; 8,383,654; 8,426,449; 8,497,381; 8,071,807; 8,193,183; 8,242,145; 8,338,484; 8,362,044; 8,378,107; 8,501,959; 8,524748; U.S. Pat Pub. Nos. 20020022218; 20030027854; 20040162323; 20040180934; 20040185509; 20040197834; 20050215609; 20070054951; 20070244131; 20070249686; 20070265278; 20070265291; 20080194600; 20080261922; 20090036469; 20090176804; 20090197959; 20100004331; 20100075990; 20100081673; 20100113503; 20100130574; 20100298368; 20110021573; 20110039852; 20110098302; 20110098352; 20110112134; 20110130453; 20110144160; 20110152338; 20110190227; 20110245303; 20110301168; 20110312974; 20110318308; 20110319445; 20120004233; 20120016029; 20120022119; 20120058123; 20120059055; 20130005728; 20130005741; 20130065902; 20130079375; 20130109685; 20130158036; each of which is expressly incorporated herein by reference. Methods of treatment and preventing preterm delivery include the administration of a therapeutic amount of prostaglandin D2 receptor antagonists to a pregnant woman in need thereof. Proper routes of administration, dosages and frequencies of administration of these tocolytic agent prostaglandin D2 receptor antagonists may be readily determined by one of skill in the art, e.g., medical practitioners.
More particularly, the present invention provides for methods of treating preterm labor via the administration of pharmaceutical compositions comprising active tocolytic therapeutic agents with a pharmaceutically acceptable carrier. Pharmaceutical composition of the present invention are intended to encompass a product comprising the active ingredient(s), e.g., prostaglandin D2 receptor antagonists, and the inert ingredient(s) (pharmaceutically acceptable excipients) that constitute the carrier, as well as any product which may result, directly or indirectly, from combination, complex formation or aggregation of any two or more of the ingredients, or from dissociation of one or more of the ingredients, or form other types of reactions or interactions of one or more of the ingredients.
For example, available prostaglandin D2 (DP1/DPGTR, DP2/CRTH2) receptor antagonists include AM156 (12′-[(cyclopropanecarbonyl-ethyl-amino)-methyl]-6-methoxy-4′-trifluoro-methyl-biphenyl-3-yl}-acetic acid, sodium salt), and AM206 (5-{2-[(benzoyloxycarbonyl-ethyl-amino)-methyl]-4-trifluoromethyl-phenyl-pyridin-3-yl)-acetic acid, sodium salt), MK-0524 ([(3R)-4-(4-Chloro-benzyl)-7-fluoro-5-(methylsulfonyl)-1,2,3,4-tetrahydrocyclopenta[b]indo1-3-yl]-acetic Acid), AM-853 (2-(4-(4-(tert-butylcarbamoyl)-2-(2-chloro-4-cyclopropylphenyl sulfonamido)phenoxy)-5-chloro-2-fluorophenyl)acetic acid), BW868C (3-benzyl-5-(6-carbohexyl)-1-(2-cyclohexyl-2-hydroxyethylamino)-hydantoin), S5751 ((Z)-7-[(1R,2R,3S,5S)-2-(5-hydroxy benzo[b]thiophen-3-ylcarbonylamino)-10-norpinan-3-yl]hept-5-enoic acid), BAY-u3405 (Ramatroban, 3(R)-[[(4-fluorophenyl) sulphonyl]aminol-1,2,3,4- tetrahydro-9H-carbazole-9-propanoic acid). Such agents, or others that are known or become known, may be used alone, in combination or subcombination. In some cases, the effects may be enhanced by selectively acting on DP1 or DP2, and therefore appropriate agents may be selected. Likewise, as may be appropriate, a mixed agonist/antagonist comprising a single or multiple agents, may be administered or concurrently administered.
For the treatment of preterm labor, the tocolytic therapeutic agents may be administered orally, by inhalation spray, topically, transdermally, parenterally or rectally in dosage unit formulations containing conventional non-toxic pharmaceutically acceptable carriers, adjuvants and vehicles. The term parenteral as used herein includes subcutaneous injections, intravenous, intramuscular, intrasternal injection or infusion techniques. Tocolytic therapeutic agents, e.g., prostaglandin D2 receptor antagonists, may be co-administered with other therapeutic agents and are suitable for simultaneous and/or sequential combination therapies. Methods of the present invention further encompass co-administration to a pregnant woman of a non-toxic therapeutically effective amount of a tocolytic therapeutic agent, such as a prostaglandin D2 receptor antagonist, optionally with other active therapeutic agents, e.g., other prostaglandin D2 receptor antagonists, either simultaneously or sequentially as part of a combination therapy treatment regimen. Similarly, a selective L-PGDS inhibitor, such as AT-56 (4-dibenzo[a,d]cyclohepten-5-ylidene-144-(2H-tetrazol-5-yl)-butyl]-piperidine) may be used alone or in combination with a DP1 and/or a DP2 antagonist. The therapeutic amounts of active therapeutic agents as administered in combination therapy may be those as commonly used for each active therapeutic agent when administered alone, or the amounts may result in lower dosage(s) for one or more of the active therapeutic agents.
As shown in
On the other hand in a ratiometric assay which seeks to determine a ratio between isoforms, or between a single isoform and the total, during purification, it may be preferred to avoid selective enrichment, and thus immunopurification may employ a non-specific antibody or a balanced mix of antibodies to ensure that the CVS sample concentrations are not altered during processing. Although the present invention has been described in relation to particular embodiments thereof, many other variations and modifications and other uses will become apparent to those skilled in the art. It is preferred, therefore, that the present invention be limited not by the specific disclosure herein, but only by the appended claims.
While preferred embodiments of the present invention have been shown and described herein, it will be obvious to those skilled in the art that such embodiments are provided by way of example only. Numerous variations, changes, and substitutions will now occur to those skilled in the art without departing from the invention. It should be understood that various alternatives to the embodiments of the invention described herein may be employed in practicing the invention. It is intended that the following claims define the scope of the invention and that methods and structures within the scope of these claims and their equivalents be covered thereby.
This application is a continuation of U.S. patent application Ser. No. 15/708,582, filed Sep. 19, 2017, now U.S. Pat. No. ______; which is a continuation of U.S. patent application Ser. No. 14/438,110, filed Apr. 23, 2015, now U.S. Pat. No. 9,797,903; which is a 371 national phase of International Application No. PCT/US2013/066490, filed Oct. 24, 2013; which claims the benefit of U.S. Provisional Patent Application 61/717,724, filed Oct. 24, 2012; the contents of which are incorporated herein by reference in their entirety.
| Number | Date | Country | |
|---|---|---|---|
| 61717724 | Oct 2012 | US |
| Number | Date | Country | |
|---|---|---|---|
| Parent | 15708582 | Sep 2017 | US |
| Child | 16523832 | US | |
| Parent | 14438110 | Apr 2015 | US |
| Child | 15708582 | US |
