JP2009501729A - Methods for the treatment and diagnosis of preterm birth complications - Google Patents

Abstract

Methods and compositions for treating and / or preventing preterm complications are provided. This method involves the administration of 2-methoxyestradiol or an analog thereof to a subject. Methods for diagnosing or predicting preterm complications are also provided. This method involves the detection of 2-methoxyestradiol or its precursors or metabolites.

Description

Background The mortality rate of extreme preterm infants is steadily decreasing, but the survival of such preterm infants is associated with considerable morbidity. The basis of care for preterm infants is to try to mimic the in utero environment with as many items as possible, to name a few: temperature regulation, nutrition, acid / base balance and oxygen supply.

  Full-term infants are exposed to elevated maternal 2ME2 levels of anti-angiogenic effects, which are numerous other pro-angiogenic events, both those produced by the mother and passed through the placenta, and those produced by the placenta itself Balanced by factors and anti-angiogenic factors. Once a fetus is delivered, it is no longer subject to either maternal or placental pro-angiogenic and anti-angiogenic factors, and the ability to produce and regulate its own factors is likely immature. Some common preterm complications such as intraventricular hemorrhage (IVH), necrotizing enterocolitis (NEC), retinopathy of prematurity (ROP), and angioma formation are angiogenesis and vascular leakage in these processes This suggests that it is caused by a lack of balance with the opposing factors.

  This imbalance of pro-angiogenic and anti-angiogenic factors appears to be occurring during the development of premature infant immature vasculature, giving balance greater significance than adults. Understanding the balance of normal angiogenesis in the uterus provides the potential for a final therapeutic intervention that complements angiogenesis-suppressing and promoting factors to mimic the normal balance of gestational age.

Overview In one aspect, this document relates to imbalances in pro-angiogenic and inhibitory factors such as intraventricular hemorrhage (IVH), necrotizing enterocolitis (NEC), retinopathy of prematurity (ROP), and hemangioma formation It features novel prevention and treatment methods and compositions that allow safer and more effective treatment of preterm infants against various disorders. In one embodiment, the method includes administering to a premature infant an effective amount of a 2-methoxyestradiol-containing composition.

  In another embodiment, the present invention develops complications due to preterm birth by assaying 2-methoxyestradiol levels in preterm infants or their mothers alone or in addition to the levels of other biomarkers It features a method for identifying preterm infants at high risk.

  Also described herein are formulations of 2-methoxyestradiol and other therapeutic agents, as well as kits, for carrying out novel diagnostic and therapeutic methods.

  These and other aspects of the invention, as well as their features and characteristics, will be apparent from the description, drawings, and claims that follow.

Detailed description
A. Definitions For convenience, certain terms employed in the specification, examples, and appended claims are collected here. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

  The articles “a” and “an” are used herein to refer to the grammatical object of one or more (ie, at least one) articles. By way of example, “an element” means one element or more than one element.

  The terms “comprise” and “comprising” are used in a comprehensive and open sense, meaning that additional elements may be included.

  The term “including” is used to mean “including but not limited to”. “Including” and “including but not limited to” are used interchangeably.

  The term “control” as used herein with respect to the level of 2-methoxyestradiol (or its precursor or metabolite) is an early term that does not suffer from at least one full-term infant or at least one preterm complication. It means the level of 2-methoxyestradiol (or its precursor or metabolite) in the baby. A control may reflect the mean level of 2-methoxyestradiol (or its precursors or metabolites) from multiple term infants or preterm infants not suffering from preterm complications. The control may be a pre-quantified level of 2-methoxyestradiol (or its precursor or metabolite) at the onset or early of a given time point. In one embodiment, the control is a graph or chart representing the control level of 2-methoxyestradiol (or its precursor or metabolite) at various gestational ages (early, full term, etc.). In one exemplary embodiment, the control represents the control level of 2-methoxyestradiol (or its precursor or metabolite) throughout all stages of pregnancy (e.g., every third trimester, monthly, weekly, or daily). It is a graph or a chart.

  As used herein, the term “premature complication” refers to any disease or disorder in an infant born before 40 weeks of gestation that is associated with an abnormal level of 2-methoxyestradiol in a subject. A disease or disorder "associated with an abnormal level of 2-methoxyestradiol" has a disease or disorder directly or indirectly caused by an abnormal level of 2-methoxyestradiol in a subject, or an independent root cause Including diseases or disorders associated with abnormal levels of 2-methoxyestradiol. Preterm birth complications can also mean a disease or disorder associated with being born 40 weeks before gestational age, in which the subject has higher levels of 2-methoxyestradiol than the control subject. Alternatively, preterm complications may refer to a disease or disorder associated with being born 40 weeks before gestational age, where the subject has lower levels of 2-methoxyestradiol than the control subject. Preterm birth complications can also mean diseases or disorders associated with abnormal angiogenesis, such as the mother, fetus, placenta, uterus. Examples of preterm complications include one or more of the following: intraventricular hemorrhage (IVH), necrotizing enterocolitis (NEC), retinopathy of prematurity (ROP), and hemangioma.

  As used herein, “diagnosis” or “diagnosing” refers to the diagnosis, prognosis, monitoring, characterization, selection or screening of patients, including participants in clinical trials, and high risk of certain disorders It includes identifying patients, patients with such disorders, or patients who are likely to respond to a particular treatment, or evaluating or monitoring a patient's response to a particular treatment.

  The term “gestational age” refers to the gestational age of the newborn, calculated from the number of weeks elapsed from the first day of the mother's last menstrual period to the date of delivery.

  The term “2-methoxyestradiol analog” is a derivative or analog of 2-methoxyestradiol that exhibits a therapeutic effect similar to that of 2-methoxyestradiol when administered to a subject suffering from a “premature complication” Means the body. In one exemplary embodiment, the 2-methoxyestradiol analog exhibits a therapeutic effect when administered to a subject suffering from preterm complications. In one embodiment, the 2-methoxyestradiol analog exhibits greater stability than 2-methoxyestradiol itself when administered to a subject. In one exemplary embodiment, the 2-methoxyestradiol analog is a compound of formula I. Examples of 2-methoxyestradiol analogs suitable for use in connection with the methods described herein include, for example, U.S. Patent No. 6,528,676, and U.S. Patent Application Nos. 2003/0236408, 2002/0147183, It has also been described in various US patents and published applications, including 2003/0187076 and 2003/0236439. In one exemplary embodiment, analogs of 2-methoxyestradiol include colchicine and combretastatin A-4.

  As used herein with respect to 2-methoxyestradiol, or 2-methoxyestradiol analog, the term “precursor” refers to 2-methoxyestradiol, or 2-methoxyestradiol analog, as at least one metabolite that occurs in the metabolic process. Means a compound that produces In one exemplary embodiment, precursors of 2-methoxyestradiol include, for example, estradiol, 2-hydroxyestradiol, and the like.

  The term “metabolite” as used herein with respect to 2-methoxyestradiol, or a 2-methoxyestradiol analog, refers to a compound produced by metabolism of 2-methoxyestradiol, or a 2-methoxyestradiol analog. Metabolites will include both anabolic and catabolic products. In one exemplary embodiment, metabolites of 2-methoxyestradiol include, for example, 2-methoxyestrone.

  The term `` biological sample '' refers to a tissue, tissue sample, or cell sample (e.g., chorionic villi sample or tissue biopsy, e.g., aspiration biopsy, brush biopsy, abrasion biopsy, needle biopsy, Punch biopsy, resected biopsy, direct biopsy, incision biopsy, or endoscopic biopsy), tumor, tumor sample, or biological fluid (e.g., blood, serum, plasma, amniotic fluid, urine) , Lymph, or cerebrospinal fluid), or a sample of biological material obtained from or present in a subject. In one exemplary embodiment, the subject can be a human subject.

  The term “approximately control level” as used herein with respect to the level of 2-methoxyestradiol and / or analogs in a subject is 50%, 30%, 25%, 20% with the control level of 2-methoxyestradiol. Means 2-methoxyestradiol levels that differ by less than 15%, 10%, 5%, 2%, or 1%.

  The term “cis” is art-recognized and refers to an arrangement of atoms or groups such that the two atoms or groups around the double bond are on the same side of the double bond. The cis configuration is often displayed as (Z) configuration.

  The term “trans” is art-recognized and refers to an arrangement of atoms or groups such that the two atoms or groups around the double bond are on opposite sides of the double bond. The transformer arrangement is often indicated as (E) arrangement.

  The term “covalent bond” is recognized in the art and electrons are electrostatically attracted to both nuclei of two atoms, and the net effect of increasing electron density between nuclei offsets the repulsion between nuclei. Means a bond between two atoms. The term covalent bond includes coordinate bonds when the bond is with a metal ion.

  The term “therapeutic agent” is art-recognized and refers to any chemical moiety that is a biological, physiological, or pharmacologically active substance that acts locally or systemically in a subject. Examples of therapeutic agents, also called “drugs”, are described in well-known references such as the Merck Index, the Physicians Desk Reference, and The Pharmacological Basis of Therapeutics, which include medical preparations; vitamins; mineral supplements; Substances used for the treatment, prevention, diagnosis, healing or alleviation of illnesses; substances that affect the structure or function of the body; or prodrugs that become biologically active or more active after being placed in a physiological environment However, it is not limited to them.

  The term “therapeutic effect” is art-recognized and refers to a local or systemic effect caused by a pharmacologically active substance in an animal, particularly a mammal, especially a human. The term “pharmacologically active substance” refers to any substance intended to be used in the diagnosis, cure, alleviation, treatment or prevention of animal or human disease or in the promotion of desirable body or mental development and / or condition. Including. The phrase “therapeutically effective amount” means the amount of such a substance that produces some desirable local or systemic effect with a reasonable profit / loss ratio applicable to any treatment. The therapeutically effective amount of such substances will vary depending on the subject being treated and the condition of the disease, the weight and age of the subject, the severity of the disease state, the mode of administration, etc. It can be easily determined if there is any. For example, certain compositions described herein may be administered in an amount sufficient to produce an effect with a reasonable profit / loss ratio applicable to such treatment.

  The term “synthetic” is art-recognized and refers to production by chemical or enzymatic synthesis in vitro.

  The term “meso compound” is art-recognized and refers to a compound that has at least two chiral centers but is achiral by a plane of symmetry or a point.

  The term “chiral” is art-recognized and refers to a molecule that has the property that it cannot be superimposed on a mirror image partner, while the term “achiral” is a molecule that can be superimposed on a mirror image partner. Means. A “prochiral molecule” is a molecule that has the potential to be converted to a chiral molecule in a particular process.

  The term “stereoisomer” is art-recognized and refers to compounds that have the same chemical structure but differ in the arrangement of atoms or groups in space, in particular, “enantiomers” overlap one another. It refers to the two stereoisomers of a compound, which cannot be mirror images. “Diastereomer”, on the other hand, refers to a stereoisomer with multiple asymmetric centers and whose molecules are not mirror images of one another.

  Furthermore, a “stereoselective process” is one in which a particular stereoisomer of a reaction product is produced in preference to other possible stereoisomers of that product. An “enantioselective process” is one that preferentially produces one of the two possible enantiomers of the reaction product.

  The term “positional isomer” is art-recognized and refers to compounds that have the same molecular formula but differ in the attachment of atoms. Thus, a “regioselective process” is one in which a specific regioisomer is produced in preference to other isomers, for example, the reaction yields a statistically significant yield of a specific regioisomer. .

  The term “epimer” is art-recognized and refers to molecules that have the same chemical structure and contain multiple stereocenters but differ in configuration at only one of these stereocenters.

  The term “ED50” is recognized in the art. In certain embodiments, ED50 means the dose of a drug that produces 50% of its maximum response or effect, or the dose that produces a predetermined response in 50% of a subject or formulation. The term “LD50” is recognized in the art. In certain embodiments, LD50 refers to the dose of a drug that is lethal in 50% of test subjects. The term “therapeutic index” is a term recognized in the art and refers to the therapeutic index of a drug defined as LD50 / ED50.

  The term “prodrug” is art-recognized and is intended to include compounds that are converted under physiological conditions to a drug, such as, for example, a 2-methoxyestradiol compound described herein. . A common method for preparing prodrugs is to select and attach moieties that are hydrolyzed under physiological conditions to provide the desired compound. In other embodiments, the prodrug is converted by the enzymatic activity of the host animal.

  The term “structure-activity relationship” or “(SAR)” is art-recognized and can be used to alter the molecular structure of a drug or other compound, such as its receptor, enzyme, nucleic acid, or other target. It means a mode in which the interaction with is changed.

  The term “aliphatic” is art-recognized and refers to a straight chain, branched, cyclic alkane, alkene, or alkyne. In certain embodiments, aliphatic groups can be straight or branched and have 1 to about 20 carbon atoms.

  The term “alkyl” is art-recognized and includes straight chain alkyl groups, branched chain alkyl groups, cycloalkyl (alicyclic) groups, alkyl substituted cycloalkyl groups, and cycloalkyl substituted alkyl groups. Contains saturated aliphatic groups. In certain embodiments, a straight chain or branched chain alkyl has no more than about 30 in its backbone (e.g., C1-C30 for straight chain, C3-C30 for branched chain), or no more than about 20 Has carbon atoms. Similarly, a cycloalkyl has about 3 to about 10 carbon atoms in its ring structure, or about 5, 6 or 7 carbons in the ring structure. The term “alkyl” is also defined to include halo-substituted alkyl.

  The term “aralkyl” is art-recognized and refers to an alkyl group substituted with an aryl group (eg, an aromatic or heteroaromatic group).

  The terms “alkenyl” and “alkynyl” are art-recognized and are similar in length and possible substitution to the alkyl described above, but each containing at least one double or triple bond. Means a group.

  Unless otherwise specified, “lower alkyl” is as defined above, but has 1 to about 10 carbons, or 1 to about 6 carbon atoms in its backbone structure. An alkyl group is meant. Similarly, “lower alkenyl” and “lower alkynyl” have similar chain lengths.

  The term “heteroatom” is art-recognized and refers to an atom of any element other than carbon or hydrogen. Exemplary heteroatoms include boron, nitrogen, oxygen, phosphorus, sulfur and selenium.

  The term “aryl” is art-recognized and is a 5-, 6-, and 7-membered monocyclic aromatic group that may contain from 0 to 4 heteroatoms, such as benzene, pyrrole, furan, thiophene. Imidazole, oxazole, thiazole, triazole, pyrazole, pyridine, pyrazine, pyridazine and pyrimidine. An aryl group having a heteroatom in the ring structure may be referred to as “heteroaryl”. An aromatic ring may be substituted at one or more positions of the ring as described above, for example, halogen, azide, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxyl, alkoxyl, amino, nitro, sulfhydryl, imino, amide, It means phosphonate, phosphinate, carbonyl, carboxyl, silyl, ether, alkylthio, sulfonyl, sulfonamide, ketone, aldehyde, ester, heterocyclyl, aromatic or heteroaromatic moiety, -CF3, -CN and the like. The term “aryl” means that multiple carbons are common to two adjacent rings (the ring is a “fused” ring), at least one of the rings is aromatic, for example, the other ring is cycloalkyl, Also included are polycyclic ring systems having multiple rings, which may be cycloalkenyl, cycloalkynyl, aryl and / or heterocyclyl.

  The terms ortho, meta and para are recognized in the art and refer to 1,2-, 1,3- and 1,4-disubstituted benzenes, respectively. For example, the names 1,2-dimethylbenzene and ortho-dimethylbenzene are synonymous.

  The term “heterocyclyl” or “heterocyclic group” is art-recognized and refers to a 3 to about 10 membered ring structure, or a 3 to 7 membered ring, wherein the ring structure contains 1 to 4 heteroatoms To do. The heterocycle may be polycyclic. Heterocyclyl groups include, for example, thiophene, thianthrene, furan, pyran, isobenzofuran, chromene, xanthene, phenoxanthene, pyrrole, imidazole, pyrazole, isothiazole, isoxazole, pyridine, pyrazine, pyrimidine, pyridazine, indolizine, isoindole, Indole, indazole, purine, quinolidine, isoquinoline, quinoline, phthalazine, naphthyridine, quinoxaline, quinazoline, cinnoline, pteridine, carbazole, carboline, phenanthridine, acridine, pyrimidine, phenanthroline, phenazine, phenalsazine, phenothiazine, furazane, phenoxazine, pyrrolidine , Oxolane, thiolane, oxazole, piperidine, piperazine, morpholine, lactone Lactams such as azetidinones and pyrrolidinones, sultams, sultones, and the like. Heterocycles are substituted at one or more positions as described above, for example, halogen, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxyl, amino, nitro, sulfhydryl, imino, amide, phosphonate, phosphinate, carbonyl, It may be substituted with carboxyl, silyl, ether, alkylthio, sulfonyl, ketone, aldehyde, ester, heterocyclyl, aromatic or heteroaromatic moiety, -CF3, -CN, and the like.

  The term “polycyclyl” or “polycyclic group” is art-recognized and includes a plurality of rings in which multiple carbons are common to two adjacent rings, eg, the ring is a “fused ring”. (Eg, cycloalkyl, cycloalkenyl, cycloalkynyl, aryl and / or heterocyclyl). Rings that are joined through non-adjacent atoms are termed “bridged” rings. Each of the polycyclic rings is substituted as described above, for example, halogen, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxyl, amino, nitro, sulfhydryl, imino, amide, phosphonate, phosphinate, carbonyl, carboxyl, silyl, It may be substituted with ether, alkylthio, sulfonyl, ketone, aldehyde, ester, heterocyclyl, aromatic or heteroaromatic moiety, -CF3, -CN, and the like.

  The term “carbocycle” is art-recognized and refers to an aromatic or non-aromatic ring in which each atom of the ring is carbon.

  The term “nitro” is art-recognized and refers to —NO 2; the term “halogen” is art-recognized and refers to —F, —Cl, —Br or —I. The term “sulfhydryl” is art-recognized and refers to —SH; the term “hydroxyl” refers to —OH; and the term “sulfonyl” is art-recognized; It means -SO2-. “Halide” refers to the corresponding anion of halogen and “pseudohalide” has the definition shown on page 560 of “Advanced Inorganic Chemistry” by Cotton and Wilkinson.

式中、R50、R51およびR52はそれぞれ独立に水素、アルキル、アルケニル、-(CH2)m-R61を意味するか、またはR50およびR51はそれらが結合しているN原子と一緒になって環構造に4から8個の原子を有する複素環を完成し；R61はアリール、シクロアルキル、シクロアルケニル、複素環または多環を意味し；かつmはゼロまたは1から8の範囲の整数である。特定の態様において、R50またはR51の一つだけがカルボニルであってもよく、例えば、R50、R51および窒素は一緒になってイミドを形成することはない。他の態様において、R50およびR51(および任意にR52)はそれぞれ独立に水素、アルキル、アルケニル、または-(CH2)m-R61を意味する。したがって、「アルキルアミン」なる用語は、それに結合している置換または無置換アルキルを有する、すなわち、R50およびR51の少なくとも一つがアルキル基である、前述の定義のアミン基を含む。 The terms “amine” and “amino” are art-recognized and refer to both unsubstituted and substituted amines, eg, moieties that can be represented by the general formula:

Wherein R50, R51 and R52 each independently represent hydrogen, alkyl, alkenyl,-(CH2) m-R61, or R50 and R51 together with the N atom to which they are attached, a ring structure Completes a heterocycle having from 4 to 8 atoms; R61 means aryl, cycloalkyl, cycloalkenyl, heterocycle or polycycle; and m is zero or an integer ranging from 1 to 8. In certain embodiments, only one of R50 or R51 may be carbonyl, for example R50, R51 and nitrogen do not combine to form an imide. In other embodiments, R50 and R51 (and optionally R52) each independently represent hydrogen, alkyl, alkenyl, or-(CH2) m-R61. Thus, the term “alkylamine” includes an amine group as defined above having a substituted or unsubstituted alkyl attached thereto, ie, at least one of R50 and R51 is an alkyl group.

式中、R50は前述の定義のとおりであり、R54は水素、アルキル、アルケニルまたは-(CH2)m-R61を意味し、ここでmおよびR61は前述の定義のとおりである。 The term “acylamino” is art-recognized and refers to a moiety that may be represented by the general formula:

In the formula, R50 is as defined above, R54 means hydrogen, alkyl, alkenyl or — (CH2) m—R61, wherein m and R61 are as defined above.

式中、R50およびR51は前述の定義のとおりである。本明細書に記載のアミドの特定の態様は、不安定でありうるイミドを含まない。 The term “amide” is art recognized as an amino-substituted carbonyl and includes a moiety that may be represented by the general formula:

In the formula, R50 and R51 are as defined above. Certain embodiments of the amides described herein do not include imides that can be unstable.

  The term “alkylthio” refers to an alkyl group as defined above having a sulfur group attached thereto. In certain embodiments, the “alkylthio” moiety is represented by one of —S-alkyl, —S-alkenyl, —S-alkynyl, and —S— (CH 2) m —R 61, wherein m and R 61 are as defined above. Defined. Representative alkylthio groups include methylthio, ethylthio, and the like.

式中、X50は結合であるか、または酸素もしくは硫黄を表し、R55およびR56は水素、アルキル、アルケニル、-(CH2)m-R61または薬学的に許容される塩を表し、R56は水素、アルキル、アルケニルまたは-(CH2)m-R61を表し、ここでmおよびR61は上で定義している。X50が酸素であり、R55またはR56が水素ではない場合、この式は「エステル」を表す。X50が酸素であり、R55が前述の定義のとおりである場合、この部分は本明細書においてカルボキシル基と呼び、特にR55が水素である場合、式は「カルボン酸」を表す。X50が酸素であり、R56が水素である場合、この式は「ギ酸エステル」を表す。一般に、上の式の酸素原子が硫黄で置き換えられた場合、式は「チオールカルボニル」基を表す。X50が硫黄であり、R55またはR56が水素ではない場合、この式は「チオールエステル」を表す。X50が硫黄であり、R55が水素である場合、式は「チオールカルボン酸」を表す。X50が硫黄であり、R56が水素である場合、この式は「チオールギ酸エステル」を表す。一方、X50が結合であり、R55が水素ではない場合、上の式は「ケトン」基を表す。X50が結合であり、R55が水素である場合、上の式は「アルデヒド」基を表す。 The term “carbonyl” is art recognized and includes a moiety that may be represented by the general formula:

Wherein X50 is a bond or represents oxygen or sulfur, R55 and R56 represent hydrogen, alkyl, alkenyl,-(CH2) m-R61 or a pharmaceutically acceptable salt, R56 represents hydrogen, alkyl , Alkenyl or — (CH 2) m —R 61, where m and R 61 are defined above. Where X50 is an oxygen and R55 or R56 is not hydrogen, the formula represents an “ester”. When X50 is oxygen and R55 is as defined above, this moiety is referred to herein as a carboxyl group, particularly where R55 is hydrogen, the formula represents a “carboxylic acid”. Where X50 is oxygen and R56 is hydrogen, this formula represents a “formate”. In general, where the oxygen atom of the above formula is replaced by sulfur, the formula represents a “thiolcarbonyl” group. Where X50 is sulfur and R55 or R56 is not hydrogen, this formula represents a “thiol ester”. Where X50 is sulfur and R55 is hydrogen, the formula represents a “thiol carboxylic acid”. Where X50 is sulfur and R56 is hydrogen, the formula represents a “thiolformate”. On the other hand, when X50 is a bond and R55 is not hydrogen, the above formula represents a “ketone” group. Where X50 is a bond, and R55 is hydrogen, the above formula represents an “aldehyde” group.

  The terms “alkoxyl” or “alkoxy” are art-recognized and refer to an alkyl group, as defined above, having an oxygen group attached thereto. Exemplary alkoxyl groups include methoxy, ethoxy, propyloxy, tert-butoxy and the like. “Ether” is two hydrocarbons covalently linked by oxygen. Thus, an alkyl such as that represented by one of -O-alkyl, -O-alkenyl, -O-alkynyl, -O-(CH2) m-R61 (m and R61 are defined above) Alkyl substituents for which is an ether are alkoxyl or similar to alkoxyl.

式中、R57は電子対、水素、アルキル、シクロアルキル、またはアリールである。 The term “sulfonate” is art-recognized and refers to a moiety that may be represented by the general formula:

Where R57 is an electron pair, hydrogen, alkyl, cycloalkyl, or aryl.

式中、R57は前述の定義のとおりである。 The term “sulfate” is art-recognized and includes a moiety that may be represented by the general formula:

In the formula, R57 is as defined above.

式中、R50およびR56は前述の定義のとおりである。 The term “sulfonamide” is art-recognized and includes a moiety that may be represented by the general formula:

In the formula, R50 and R56 are as defined above.

式中、R50およびR51は前述の定義のとおりである。 The term “sulfamoyl” is art-recognized and refers to a moiety that may be represented by the general formula:

In the formula, R50 and R51 are as defined above.

式中、R58は下記の一つである：水素、アルキル、アルケニル、アルキニル、シクロアルキル、ヘテロシクリル、アリールまたはヘテロアリール。 The term “sulfonyl” is art-recognized and refers to a moiety that may be represented by the general formula:

Wherein R58 is one of the following: hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl or heteroaryl.

式中、R58は上で定義している。 The term “sulfoxide” is art-recognized and refers to a moiety that may be represented by the general formula:

Where R58 is defined above.

式中、Q50はSまたはOを表し、R59は水素、低級アルキルまたはアリールを表す。例えば、アルキルを置換するために用いる場合、ホスホリルアルキルのホスホリル基は下記の一般式で表しうる：

式中、Q50およびR59はそれぞれ独立に上で定義しており、Q51はO、SまたはNを表す。Q50がSである場合、ホスホリル部分は「ホスホロチオエート」である。 The term “phosphoryl” is art-recognized and may generally be represented by the following formula:

In the formula, Q50 represents S or O, and R59 represents hydrogen, lower alkyl or aryl. For example, when used to substitute alkyl, the phosphoryl group of phosphorylalkyl may be represented by the general formula:

Wherein Q50 and R59 are each independently defined above and Q51 represents O, S or N. When Q50 is S, the phosphoryl moiety is “phosphorothioate”.

式中、Q51、R50、R51およびR59は前述の定義のとおりである。 The term “phosphoramidite” is art-recognized and may be represented by the general formula:

In the formula, Q51, R50, R51 and R59 are as defined above.

式中、Q51、R50、R51およびR59は前述の定義のとおりであり、R60は低級アルキルまたはアリールを表す。 The term “phosphonamidite” is art recognized and may be represented by the general formula:

In the formula, Q51, R50, R51 and R59 are as defined above, and R60 represents lower alkyl or aryl.

  Similar substitutions may be made on alkenyl and alkynyl groups to produce, for example, aminoalkenyl, aminoalkynyl, amidoalkenyl, amidoalkynyl, iminoalkenyl, iminoalkynyl, thioalkenyl, thioalkynyl, carbonyl-substituted alkenyl or alkynyl.

  When each expression, eg, alkyl, m, n, etc., occurs more than once in any structure, the definition is intended to be independent of its definition elsewhere in the same structure.

  The term “selenoalkyl” is art-recognized and refers to an alkyl group having a substituted seleno group attached thereto. Exemplary “selenoethers” that can be substituted on alkyl are —Se-alkyl, —Se-alkenyl, —Se-alkynyl, and —Se— (CH 2) m —R 61, where m and R 61 are defined above. ).

  The terms trifuryl, tosyl, mesyl, and nonafryl are recognized in the art and refer to trifluoromethanesulfonyl, p-toluenesulfonyl, methanesulfonyl, and nonafluorobutanesulfonyl groups, respectively. The terms triflate, tosylate, mesylate, and nonaflate are recognized in the art and are trifluoromethanesulfonate, p-toluenesulfonate, methanesulfonate, and nonafluorobutanesulfonate functional groups, respectively, Means a molecule containing a group.

  The abbreviations Me, Et, Ph, Tf, Nf, Ts, and Ms represent methyl, ethyl, phenyl, trifluoromethanesulfonyl, nonafluorobutanesulfonyl, p-toluenesulfonyl, and methanesulfonyl, respectively. A more comprehensive list of abbreviations used by those skilled in organic chemistry is given in the first issue of each volume of the Journal of Organic Chemistry; this list is typically shown in the table titled Standard List of Abbreviations Yes.

  Certain compounds included in the compositions described herein may exist in particular geometric or stereoisomeric forms. In addition, the polymers described herein may be optically active. All such compounds, including cis and trans isomers, R- and S-enantiomers, diastereomers, (D) -isomers, (L) -isomers, racemic mixtures thereof, and other mixtures Is contemplated herein. Additional asymmetric carbon atoms may be present in substituents such as alkyl groups. All such isomers, as well as mixtures thereof, are intended to be included in the compositions described herein.

  For example, if a particular enantiomer of a compound described herein is desired, it may be prepared by asymmetric synthesis or by induction with a chiral auxiliary, in which case The resulting diastereomeric mixture is separated and the auxiliary group is cleaved to give the pure desired enantiomer. Or, if the molecule contains a basic functional group such as amino, or an acidic functional group such as carboxyl, a diastereomeric salt formed after forming a diastereomeric salt with an appropriate optically active acid or base Is resolved by fractional crystallization or chromatographic means well known in the art followed by recovery of the pure enantiomer.

  “Substituted” or “substituted” is consistent with the atom to which such substitution is substituted and the allowed valency of the substituent, and is stable by substitution, eg, rearrangement, cyclization, desorption. It is understood to include implicit conditions that result in compounds that do not spontaneously undergo conversion, such as by separation or other reactions.

  The term “substituted” is also intended to include all permissible substituents of organic compounds. In a broad aspect, acceptable substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and non-aromatic substituents of organic compounds. Exemplary substituents include, for example, those previously described herein. The permissible substituents can be one or more and the same or different for appropriate organic compounds. In certain embodiments, a heteroatom such as nitrogen may have a hydrogen substituent and / or any acceptable substituent of the organic compounds described herein that satisfy the valence of the heteroatom. The permissible substituents of organic compounds are not limited in any way by this description.

  Chemical elements can be identified according to the Periodic Table of the Elements, CAS version, Handbook of Chemistry and Physics, 67th Ed., 1986-87, inside cover. In addition, the term “hydrocarbon” is intended to include all acceptable compounds having at least one hydrogen and one carbon atom. In a broad aspect, acceptable hydrocarbons include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and non-aromatic organic compounds that can be substituted or unsubstituted. It is.

  The term “protecting group” is art-recognized and refers to a temporary substituent that protects a potentially reactive functional group from undesired chemical transformations. Examples of such protecting groups include esters of carboxylic acids, silyl ethers of alcohols, and acetals and ketals of aldehydes and ketones, respectively. The field of protecting group chemistry is reviewed by Greene and Wuts in Protective Groups in Organic Synthesis (2nd ed., Wiley: New York, 1991).

  The term “hydroxyl protecting group” is art-recognized and refers to a group intended to protect a hydroxyl group against undesired reactions during synthetic procedures, such as benzyl or in the art. Other known suitable ester or ether groups are included.

  The term “carboxyl protecting group” is art-recognized and protects a carboxylic acid group, such as an amino acid or peptide C-terminus or an acidic or hydroxyl azepine ring substituent, against undesired reactions during synthetic procedures. Means and includes groups intended to Examples of the protecting group for carboxyl group include, for example, benzyl ester, cyclohexyl ester, 4-nitrobenzyl ester, t-butyl ester, 4-pyridylmethyl ester and the like.

  The term “amino protecting group” is art-recognized and prevents the amino group from participating in reactions performed on other functional groups, but can be removed from the amine if desired. Means a group. Such groups are discussed by Greene and Wuts, Ch. 7, cited above, and Barton, Protective Groups in Organic Chemistry ch. 2 (McOmie, ed., Plenum Press, New York, 1973). Examples of suitable groups include, by way of example, formyl, dansyl, acetyl, benzoyl, trifluoroacetyl, succinyl, methoxysuccinyl, benzyl and 3,4-dimethoxybenzyl, o-nitrobenzyl, and triphenylmethyl. Acyl protecting groups such as substituted benzyl; formula —COOR (where R is methyl, ethyl, propyl, isopropyl, 2,2,2-trichloroethyl, 1-methyl-1-phenylethyl, isobutyl, t-butyl, t-amyl; , Vinyl, allyl, phenyl, benzyl, p-nitrobenzyl, o-nitrobenzyl, and 2,4-dichlorobenzyl); formyl, acetyl, chloroacetyl, dichloroacetyl, trichloroacetyl, tri Acyl groups and substituted acyls such as fluoroacetyl, benzoyl, and p-methoxybenzoyl; Methanesulfonyl, p- toluenesulfonyl, p- bromobenzenesulfonyl, p- nitrophenyl ethyl, and p- toluenesulfonyl - include other groups such as aminocarbonyl. Preferred amino protecting groups are benzyl (-CH2C6H5), acyl [C (O) R1] or SiR13 (R1 is C1-C4 alkyl, halomethyl, or 2-halo-substituted- (C2-C4 alkoxy), such as carbonylbenzyloxy ( Aromatic urethane groups such as Cbz); and aliphatic urethane protecting groups such as t-butyloxycarbonyl (Boc) or 9-fluorenylmethoxycarbonyl (FMOC).

  When each expression, eg, lower alkyl, m, n, p, etc., occurs more than one time in any structure, the definition is intended to be independent of its definition elsewhere in the same structure.

  The term “electron withdrawing group” is art-recognized and refers to the tendency of a substituent to attract valence electrons from an adjacent atom, ie, the substituent is electronegative with respect to the adjacent atom. Quantification of the level of electron withdrawing ability is obtained by Hammett sigma (σ) constant. This well-known constant is described in many references, for example, March, Advanced Organic Chemistry 251-59 (McGraw Hill Book Company: New York, 1977). Hammett constant values are generally negative for electron donating groups (σ (P) =-0.66 for NH2), positive for electron withdrawing groups (σ (P) = 0.78 for nitro groups), and σ (P) is Indicates para substitution. Exemplary electron withdrawing groups include nitro, acyl, formyl, sulfonyl, trifluoromethyl, cyano, chloride, and the like. Exemplary electron donating groups include amino, methoxy and the like.

  The term `` modulation '' when used in reference to a functional or biological activity or process (e.g., enzyme activity or receptor binding), up-regulate (e.g., activate or stimulate), down-regulate. (Eg, inhibit or suppress) or otherwise the ability to change the quality of such a property, activity or process. In certain cases, such regulation may depend on the appearance of certain events, such as activation of signal transduction pathways, and / or may appear only in certain cell types.

  The term “treating” is art-recognized and refers to curing as well as ameliorating at least one symptom of any condition or disease. In one exemplary embodiment, treating a complication of preterm birth includes, for example, inhibiting the disease or disorder, for example, stopping its onset, or ameliorating or reducing at least one symptom of the disease or disorder. For example, causing regression of complications.

  The term “prophylactic” or “therapeutic” treatment of preterm complications refers to the administration of one or more of the compositions of the invention to a host. When administered prior to clinical manifestation of an undesired condition (e.g., host animal disease or other undesired condition), the treatment is prophylactic, i.e., against the occurrence of an undesired condition for the host Where administered, treatment is therapeutic when administered after the onset of an undesired condition (i.e. intended to reduce, ameliorate or maintain an existing undesired condition or its side effects) .

  “Patient”, “subject” or “host” to be treated by the methods of the present invention can mean either a human or non-human animal.

  The term “mammal” is known in the art and exemplary mammals include humans, primates, cows, pigs, dogs, cats, and rodents (eg, mice and rats).

  The term “bioavailable” in the context of a compound is art-recognized and is absorbed, taken up, or otherwise physiologically available to the subject or patient to whom the compound is administered. Means the form of the compound or part of the dosage.

  The term “pharmaceutically acceptable salt” is art-recognized and includes, for example, relatively non-toxic inorganic and organic acid additions of compounds, including those included in the compositions described herein. Means salt.

  The term “pharmaceutically acceptable carrier” is art-recognized and allows any composition of the invention or component thereof to be transferred from one organ or body part to another organ or body part. It means a pharmaceutically acceptable material, composition or vehicle, such as a liquid or solid filler, diluent, excipient, solvent or encapsulating material involved in carrying or transporting to a part. Each carrier must be “acceptable” in the sense of being compatible with the composition of the invention and its components and not injurious to the patient. Some examples of materials that can serve as pharmaceutically acceptable carriers include: (1) sugars such as lactose, glucose and sucrose; (2) starches such as corn starch and potato starch; (3) Cellulose and its derivatives such as sodium carboxymethylcellulose, ethylcellulose and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) talc; (8) excipients such as cocoa butter and suppository wax (9) oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; (10) glycols such as propylene glycol; (11) polyols such as glycerin, sorbitol, mannitol and polyethylene glycol; 12) Esters such as ethyl oleate and ethyl laurate; (13) Agar; (14) buffering agents such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer's solution; (19) ethyl alcohol; (20) phosphate buffer solution; and (21) other non-toxic compatible materials used in pharmaceutical formulations.

  The terms “systemic administration”, “administered systemically”, “peripheral administration”, and “administered peripherally” are art-recognized and the composition, therapeutic agent or other material of the present invention is By its administration other than directly into the central nervous system, eg, subcutaneous administration, which enters the patient's system and is thus susceptible to metabolism and other similar processes.

  The terms “parenteral administration” and “administered parenterally” are art-recognized and refer to modes of administration other than enteral and topical administration, usually by injection, intravenous, intramuscular, Intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transorgan, subcutaneous, epidermal, intraarticular, subcapsular, subarachnoid, intrathecal, and intrasternal injection and infusion Including, but not limited to.

  Contemplated equivalents of the compositions described herein are subject to one or more simple changes in substituents or components that do not deleteriously affect the characteristics of the composition of interest. Except for compositions having the same general properties. In general, the components of the compositions described herein can be prepared using readily available starting materials, reagents, and conventional synthetic methods. In these reactions, variants that are known per se but are not mentioned here can also be used.

B. Treatment, Prevention and Diagnostic Methods for Preterm Complications Provided herein are methods and compositions for treating, preventing and / or diagnosing preterm complications. In one embodiment, methods and compositions for treating or preventing preterm complications using 2-methoxyestradiol or analogs thereof are provided. In another embodiment, a method is provided for diagnosing preterm complications by quantifying the level of 2-methoxyestradiol (or a precursor or metabolite thereof) in a subject. In one exemplary embodiment, the methods and compositions described herein can be used to reduce premature complications by administering 2-methoxyestradiol or an analog of 2-methoxyestradiol to a subject in need thereof. It may be used to treat or prevent. In another exemplary embodiment, the methods and compositions described herein can be used to diagnose premature birth complications, or the likelihood of developing preterm birth complications, in a subject, such as 2-methoxyestradiol (or its). It may be used for diagnosis including detecting the level of precursors or metabolites).

  In certain embodiments, the methods described herein can include first identifying a subject that is susceptible to or suffering from preterm complications. Subjects who are susceptible to or suffering from preterm complications may be identified by recognizing symptoms of preterm complications presented by the patient. Alternatively, subjects prone to or suffering from preterm complications are identified by quantifying the level of 2-methoxyestradiol (or its precursor or metabolite) in the subject and comparing it to a control. May be. Any deviation from the control level of 2-methoxyestradiol (or its precursor or metabolite) may be indicative of a subject suffering from preterm complications. In one exemplary embodiment, a subject is periodically (or at regular intervals) 2-methoxyestradiol (or at regular intervals) to diagnose preterm complications or to monitor the stage or onset of preterm complications. It may be screened for levels of its precursors or metabolites). In one embodiment, screening for levels of 2-methoxyestradiol (or its precursors or metabolites) is performed regularly, e.g., approximately once a month, once every three weeks, once every two weeks, 10 days. It may be performed once, once a week, or approximately once every 144, 120, 96, 72, 48, 24, or 12 hours. In one exemplary embodiment, screening for levels of 2-methoxyestradiol (or precursors or metabolites thereof) may be used to identify subjects that may be candidates for treatment with 2-methoxyestradiol or analogs thereof. Good. In another embodiment, screening for levels of 2-methoxyestradiol (or precursors or metabolites thereof) may be used to calculate a dose effective for administration to a subject.

  A subject identified as having a low or high level of 2-methoxyestradiol relative to controls may be prone to or suffering from preterm complications. In one embodiment, a subject having a low level of 2-methoxyestradiol compared to a control may be susceptible to or suffer from one or more of the following preterm complications: intraventricular Bleeding (IVH), necrotizing enterocolitis (NEC), retinopathy of prematurity (ROP), and hemangioma formation.

  The degree or severity of preterm complications may be assessed based on the degree of bias in the subject's level of 2-methoxyestradiol compared to controls. For example, subjects with a greater bias in 2-methoxyestradiol levels compared to controls may indicate that they are more likely to suffer from or suffer from more severe cases of preterm complications.

  Screening for levels of 2-methoxyestradiol (or its precursors or metabolites) may be used to monitor the course of treatment with 2-methoxyestradiol or its analogs. When treating a subject with an analog of 2-methoxyestradiol, the level of 2-methoxyestradiol (or its precursor or metabolite) and / or the level of 2-methoxyestradiol analog (or its precursor or metabolite) Quantifying can be useful for monitoring. In one exemplary embodiment, it may be useful to quantify the level of 2-methoxyestradiol and the level of 2-methoxyestradiol analog being administered to a subject. In another embodiment, it may be useful to quantify the level of precursor of 2-methoxyestradiol and the level of precursor of 2-methoxyestradiol analog. In yet another embodiment, it may be useful to quantify the level of 2-methoxyestradiol metabolite and the level of 2-methoxyestradiol analog metabolite.

  The level of 2-methoxyestradiol in a subject may be directly quantified by measuring the level of 2-methoxyestradiol itself. Alternatively, the level of 2-methoxyestradiol may be quantified indirectly by measuring the level of a precursor or metabolite of 2-methoxyestradiol. Similarly, the level of an analog of 2-methoxyestradiol may be quantified by directly measuring the level of the analog in the subject, or indirectly by measuring the level of an analog precursor or metabolite. May be quantitatively determined. In yet another embodiment, the level of 2-methoxyestradiol or its analog precursor or metabolite may be measured directly for diagnosis or monitoring. In this case, the precursor or analyte is not used as a surrogate for the level of 2-methoxyestradiol or its analog, but rather is used directly for diagnosis or monitoring.

  The level of 2-methoxyestradiol or an analog thereof may be quantified in a biological sample of a subject or a control subject. In one exemplary embodiment, the level of 2-methoxyestradiol may be quantified in a urine, blood or plasma sample from the subject.

  The level of 2-methoxyestradiol or an analog thereof in a subject's biological sample may be directly compared to a control. In another embodiment, the level of 2-methoxyestradiol or an analog thereof in a biological sample of the subject may be used to calculate the physiological concentration of 2-methoxyestradiol or an analog found in the subject . The physiological concentration of 2-methoxyestradiol or an analog thereof in the subject may then optionally be compared to the control.

  The level of 2-methoxyestradiol (or a precursor or metabolite thereof) or an analogue of 2-methoxyestradiol (or a precursor or metabolite thereof) in a sample, for example, cord blood, amniotic fluid, plasma, breast milk, etc. Quantification may be performed using any method known in the technical field. For example, the level of 2-methoxyestradiol and / or its analogs may be measured using gas chromatography-mass spectrometry (GC-MS), which can be used to measure 2ME2 in cord blood, amniotic fluid, plasma, breast milk, etc. Or it may be used to measure the level of its analogs. GC-MS is a standard test method for detecting and quantifying low molecular weight steroids such as 2-methoxyestradiol and / or its analogs. In another exemplary embodiment, the level of 2-methoxyestradiol and / or its analog (and its precursor or metabolite) in the sample is measured by thin layer chromatography (TLC), high performance liquid chromatography (HPLC), etc. It may be quantified using other chromatographic methods. Alternatively, the level of 2-methoxyestradiol and / or its analog (and its precursor or metabolite) may be quantified using immunoassay methods such as ELISA, RIA.

  Furthermore, a method for detecting precursors of 2-methoxyestradiol, such as estrogen and estradiol (see BT Zhu and AH Conney, Cancer Research (1998) 58: 2269 for a review of pathways to 2-methoxyestradiol). Good. For example, serum estrogen assays, such as the commercially available hemagglutination inhibition kit (Hi-Estrotec) originally designed to measure total urinary estrogen, with a minimum sensitivity of 17-beta-estradiol 100 pg and less than 2.5 hours May be used to modify hemagglutination inhibition assays, such as the rapid, sensitive and simple non-radioactive serum estrogen assay developed by Kaseki et al. Another example of an estrogen assay that can be used in the methods of the present invention is the Applied Biosystems trademark HitHunter Enzyme Fragment Complementation (EFC) assay. This assay is based on an engineered β-galactosidase enzyme consisting of two fragment-enzyme acceptors (EA) and an enzyme donor (ED). When the fragments are separated, there is no β-galactosidase activity, but when the fragments are brought together, they bind (complement) to form an active enzyme. EFC assays use ED-analyte conjugates where the analyte can be recognized by a specific binding protein such as an antibody or receptor. In the absence of a specific binding protein, the ED-analyte conjugate can complement EA to form active β-galactosidase, producing a positive luminescent signal. When a specific binding protein binds to the ED-analyte conjugate, complementation with EA is prevented and no signal is generated. When free analyte is provided (in the sample), this will compete with the ED-analyte conjugate for binding to a specific binding protein; the free analyte will complement the EA. Will release the ED-analyte conjugate and produce a signal depending on the amount of free analyte in the sample.

  The level of 2-methoxyestradiol or a precursor or analog thereof in a subject may be compared quantitatively or qualitatively with a control. For example, a qualitative (or unitless) comparison may be performed by assessing whether the level of 2-methoxyestradiol or its analog in a subject is higher, lower, or about the same as a control. Good. Optionally, a qualitative comparison is used to estimate the degree of difference in the level of 2-methoxyestradiol or its analog in the subject, for example, a 2-fold change, 50% change, etc. compared to the control. May be. A quantitative comparison may be performed by assessing the amount of 2-methoxyestradiol or an analog thereof in a subject compared to the amount in a control, where the amount is in some form of unit. (Eg, protein mg, amount of spots / bands in the gel, spot intensity of fluorescence imaging device or autoradiogram exposure, amount of spots on the chromatography plate, etc.).

  In addition to or instead of monitoring the level of 2-methoxyestradiol (or its precursor or metabolite) in a subject, it may be desirable to monitor the symptoms of preterm complications. For example, in a subject suffering from or susceptible to retinopathy of prematurity, performing an eye examination of the subject before, during and / or after treatment with 2-methoxyestradiol or an analogue thereof of the present invention May be desirable. After dropping a series of extended eye drops into each eye, the retina is examined with an ophthalmoscope. For example, performing cranial ultrasound of a subject in a subject suffering from or susceptible to intracerebroventricular hemorrhage before, during and / or after treatment with 2-methoxyestradiol or an analog thereof of the present invention May be desirable. This test uses sound waves to create an image of the internal structure. Cranial ultrasound can be seen inside the infant's brain through the fountain, the space between the infant's head bones. Ultrasound can grade the amount of bleeding. Abdominal x-ray and / or blood test of a subject suffering from or susceptible to necrotizing enterocolitis before, during and / or after treatment with 2-methoxyestradiol or an analog thereof of the present invention It may be desirable to do this. X-rays of the abdomen of subjects with necrotizing enterocolitis may show signs of foam in the intestine and signs of air or gas in the hepatic vena cava. Air may also be found outside the abdominal intestines. A needle may be inserted into the abdominal cavity. Collecting intestinal fluid from the abdomen is often a sign that there is a hole in the intestine. Physical examination, CT scan and / or MRI of the subject before, during and / or after treatment with 2-methoxyestradiol or an analogue thereof in a subject suffering from or susceptible to hemangiomas It may be desirable to perform a scan.

  In one embodiment, the method described herein includes administering 2-methoxyestradiol to the subject. Alternatively, it may be desirable to administer to a subject a mixture of 2-methoxyestradiol and one or more analogs of 2-methoxyestradiol, or a mixture of analogs of 2-methoxyestradiol. In other embodiments, 2-methoxyestradiol and / or analogs thereof are administered to a subject in combination with another therapeutic agent effective to treat or ameliorate at least one symptom of preterm complications. May be desirable. In one exemplary embodiment, 2-methoxyestradiol is another treatment effective to treat or ameliorate preterm complications, such as, for example, a lung surfactant to help the lungs function normally. Administer with drugs. Exemplary pulmonary surfactants include, but are not limited to, corphosseryl (registered trademark Exosurf), registered trademark Survanta, and registered trademark Surfaxin. In embodiments directed to combination therapy, the drugs may be formulated and administered together as a mixture or may be formulated and administered separately.

  In one exemplary embodiment, a subject susceptible to or suffering from preterm complications associated with low levels of 2-methoxyestradiol is treated with a level of 2-methoxyestradiol (or a level of 2-methoxyestradiol + 2 A sufficient amount of 2-methoxyestradiol or an analog thereof is administered to increase the level of -methoxyestradiol analog) to approximately the same level as the control subject's 2-methoxyestradiol level. In a further embodiment, the subject has an amount sufficient to maintain the subject's 2-methoxyestradiol level (or 2-methoxyestradiol level + 2-methoxyestradiol analog level) at about the same level as the control level. 2-methoxyestradiol or an analog thereof may be administered periodically. For example, in certain embodiments, it may be desirable to administer 2-methoxyestradiol and / or analogs thereof to a subject monthly, weekly, or daily. In one exemplary embodiment, the level of 2-methoxyestradiol (or the level of 2-methoxyestradiol + the level of 2-methoxyestradiol analog) is maintained in the subject at about the same level as the control level throughout pregnancy. To maintain the subject's appropriate level of 2-methoxyestradiol (or the level of 2-methoxyestradiol + the level of 2-methoxyestradiol analog), the level of 2-methoxyestradiol (or the level of 2-methoxyestradiol + 2- It may be desirable to monitor the subject regularly for levels of methoxyestradiol analog) and adjust the amount of therapeutic agent administered to the subject accordingly.

式中、それぞれの出現について独立に：
RはH、C1-6アルキル、アリール、アラルキル、またはカルボニルであり；
R1およびR2は-OR、-SR、または-N(R)2であり；
R3はH、ハロゲン化物、C1-6アルキル、アリール、アラルキル、またはカルボニルであり；
R4、R5、およびR6はH、ハロゲン化物、C1-6アルキル、アリール、アラルキル、またはカルボニルであり；
mは1〜5までの整数であり；かつ
nおよびpは1〜6までの整数である。 In one embodiment, 2-methoxyestradiol analogs suitable for use in accordance with the methods and compositions described herein include compounds of formula I:

In the formula, for each occurrence independently:
R is H, C1-6 alkyl, aryl, aralkyl, or carbonyl;
R1 and R2 are -OR, -SR, or -N (R) 2;
R3 is H, halide, C1-6 alkyl, aryl, aralkyl, or carbonyl;
R4, R5, and R6 are H, halide, C1-6 alkyl, aryl, aralkyl, or carbonyl;
m is an integer from 1 to 5; and
n and p are integers from 1 to 6.

  In further embodiments, 2-methoxyestradiol analogs suitable for use in accordance with the methods and compositions described herein include compounds of formula I and the attendant definitions, wherein R is methyl.

  In further embodiments, 2-methoxyestradiol analogs suitable for use in accordance with the methods and compositions described herein include compounds of formula I and the attendant definitions, wherein R1 is OH.

  In further embodiments, 2-methoxyestradiol analogs suitable for use in accordance with the methods and compositions described herein include compounds of formula I and the attendant definitions, wherein R2 is OH.

  In further embodiments, 2-methoxyestradiol analogs suitable for use in accordance with the methods and compositions described herein include compounds of formula I and the attendant definitions, wherein R3 is methyl.

  In further embodiments, 2-methoxyestradiol analogs suitable for use in accordance with the methods and compositions described herein include compounds of formula I and the attendant definitions, wherein R4 is H.

  In further embodiments, 2-methoxyestradiol analogs suitable for use in accordance with the methods and compositions described herein include compounds of formula I and the attendant definitions, wherein R5 is H.

  In further embodiments, 2-methoxyestradiol analogs suitable for use in accordance with the methods and compositions described herein include compounds of formula I and the attendant definitions, wherein R6 is H.

  In further embodiments, 2-methoxyestradiol analogs suitable for use according to the methods and compositions described herein include compounds of formula I and the attendant definitions, wherein R is methyl and R1 is OH. .

  In further embodiments, 2-methoxyestradiol analogs suitable for use in accordance with the methods and compositions described herein include compounds of formula I and the attendant definitions, wherein R is methyl and R2 is OH. .

  In further embodiments, 2-methoxyestradiol analogs suitable for use in accordance with the methods and compositions described herein include compounds of formula I and the attendant definitions, wherein R is methyl and R3 is methyl. .

  In further embodiments, 2-methoxyestradiol analogs suitable for use in accordance with the methods and compositions described herein include Formula I and the attendant wherein R is methyl, R1 is OH, and R2 is OH. Of the definition of

  In further embodiments, 2-methoxyestradiol analogs suitable for use in accordance with the methods and compositions described herein are those wherein R is methyl, R1 is OH, R2 is OH, and R3 is methyl. Including compounds of formula I and the attendant definitions.

  In further embodiments, 2-methoxyestradiol analogs suitable for use in accordance with the methods and compositions described herein include compounds of formula I and the attendant definitions, wherein R4, R5, and R6 are H.

  In further embodiments, 2-methoxyestradiol analogs suitable for use in accordance with the methods and compositions described herein are those of formula I and the attendant wherein R is methyl and R4, R5, and R6 are H. Includes defined compounds.

  In a further embodiment, a 2-methoxyestradiol analog suitable for use in accordance with the methods and compositions described herein is wherein R is methyl, R1 is OH, and R4, R5, and R6 are H. Including certain compounds of formula I and the attendant definitions.

  In further embodiments, 2-methoxyestradiol analogs suitable for use in accordance with the methods and compositions described herein are those wherein R is methyl, R1 is OH, R2 is OH, and R4, R5 And the compounds of formula I and the attendant definitions, wherein R 6 is H.

  In a further embodiment, a 2-methoxyestradiol analog suitable for use in accordance with the methods and compositions described herein is R is methyl, R1 is OH, R2 is OH, and R3 is methyl. And compounds of formula I and the attendant definitions, wherein R4, R5, and R6 are H.

  Also included in the compositions and methods described herein are pharmaceutically acceptable addition salts and complexes of Formula I. Where a compound can have one or more chiral centers, unless stated otherwise, the compositions described herein include each unique racemic compound as well as each unique non-racemic compound.

および

などの、ケト-エノール互変異性体などの互変異性型で存在しうる場合、それぞれの互変異性型は、平衡状態またはR'による適当な置換により一つの型にロックされた状態のいずれで存在しようと、本明細書に記載の化合物と関連して含まれることが意図される。任意の一つの出現における任意の置換基の意味は、任意の他の出現におけるその意味または任意の他の置換基の意味とは独立である。 Where a compound has an unsaturated carbon-carbon double bond, both cis (Z) and trans (E) isomers are intended to be included in connection with the compounds described herein. Compound is

and

Such as keto-enol tautomers, each tautomeric form is either in equilibrium or locked into one form by appropriate substitution with R ′. Is intended to be included in connection with the compounds described herein. The meaning of any substituent at any one occurrence is independent of its meaning at any other occurrence or the meaning of any other substituent.

  Prodrugs of 2-methoxyestradiol and compounds of Formula I are also included in the methods and compositions described herein.

  Compounds of formula I may be prepared by any conventional method useful for the preparation of similar compounds. The starting materials for this step are known or can be prepared by known steps from commercially available materials. A compound used in the methods described herein can be converted to another compound used in the methods described herein using conventional methods. The product of the reaction is isolated by conventional means such as extraction, crystallization, distillation, chromatography and the like.

  Known compounds used in accordance with the present invention and precursors to the novel compounds of the present invention can be purchased, for example, from Sigma Chemical Co., St. Louis, Steraloids and Research Plus. Other compounds of the present invention can be synthesized from publicly available precursors according to known methods.

  The chemical synthesis of estradiol has been described (Eder, V. et al., Ber 109, 2948 (1976); Oppolzer, DA and Roberts, DA Helv. Chim. Acta. 63, 1703, (1980)). The synthetic route used to prepare some of the 2-methoxyestradiol analogs of the present invention is based on modified published literature methods for estradiol derivatives and dimethylhydrazone (Trembley et al., Bioorganic & Med Chem. 1995 3, 505-523; Fevig et al., J. Org. Chem., 1987 52, 247-251; Gonzalez et al., Steroids 1982, 40, 171-187; Trembley et al., Synthetic Communications 1995, 25, 2483-2495; Newkome et al., J. Org. Chem. 1966, 31, 677-681; Corey et al Tetrahedron Lett 1976, 3-6; Corey et al., Tetrahedron Lett, 1976, 3667- 3668) and German Patent No. 2757157 (1977).

  In exemplary embodiments, 2-methoxyestradiol analogs suitable for use with the compositions and methods described herein are US Pat. No. 6,528,676, and US Patent Applications 2003/0236408, 2002/0147183. 2003/0187076, 2002/0082433, and 2003/0236439. 2-Methoxyestradiol or analogs thereof may be purchased from commercially available suppliers or, for example, U.S. Patent No. 6,528,676 and U.S. Patent Application Nos. 2003/0236408, 2002/0147183, 2003. It may be prepared as described in / 0187076, 2002/0082433, and 2003/0236439. Examples of 2-methoxyestradiol analogs suitable for use in accordance with the methods and compositions described herein include one or more of the following: 17-substituted 2-methoxyestradiol derivatives: estra-1,3, 5 (10) -triene-3,17α-diol, 2-methoxyestradi-1,3,5 (10) -trien-3-ol, 17β-aminoestradi-1,3,5, (10) -triene- 3,17β-diol, 2-methoxy-17-oxime-3-hydroxyestradi-1,3,5 (10) -trien-17-one, 2-methoxy-3,17β-bis (acetyloxy) estradi-1 , 3,5, (10) -triene, 2-methoxy-17β-propaneestradi-1,3,5 (10) -trien-3-ol, 2-methoxy-17β-methylestradi-1,3,5 ( 10) -Trien-3-ol, 2-methoxy-17 (20) -Z-propylideneestradi-1,3,5, (10) -trien-3-ol, 17 (20) -methyleneestradi-1, 3,5 (10) -trien-3-ol, 2-methoxy-17β- (Nn- (1) propylamino ) Estra-1,3,5 (10) -trien-3-ol, 2-methoxy-19-norpregna-1,3,5 (10) 17 (20) -tetraen-3-ol, 2-methoxy-17β -Ethyl estra-1,3,5 (10) -trien-3-ol, 2-methoxy-17- (4-tosylhydrazone) estra-1,3,5 (10) -trien-3-ol, 2- Methoxy-17 (20) -Z-butylideneestradi-1,3,5 (10) -trien-3-ol, 2-methoxy-17β-butylestradi-1,3,5 (10) -triene-3- All: Disubstituted estradiol derivatives: 2-acetylestradi-1,3,5 (10) -triene-3,17β-diol, 2-formylestradi-1,3,5 (10) -triene-3,17β-diol 2- (hydroxymethyl) estradi-1,3,5 (10) -triene-3,17β-diol, 2-ethylestradi-1,3,5 (10) -triene-3,17β-diol, 2- Methyl estra-1,3,5 (10) -triene-3,17β-diol, 2-nitroestradi-1,3,5 (10) -triene-3,17β-diol, 2- (N, N-dimethyl Amino) Estra -1,3,5 (10) -triene-3,17β-diol, 2-aminoestradi-1,3,5 (10) -trien-3-ol, 2-formamide estra-1,3,5 (10 ) -Trien-3-ol, 2- (N-methylamino) estradi-1,3,5 (10) -trien-3-ol-HCl, 2- (N, N-dimethylamino) estradi-1,3 , 5 (10) -Trien-3-ol, 2- (N, N-dimethylamino) estradi-1,3,5 (10) -trien-3-ol-HCl, 2-aminoestradi-1,3, 5 (10) -triene-3,17β-diol, 2- (N, N-dimethylamino) -17 (20) -methyleneestradi-1,3,5 (10) -trien-3-ol-HCL, 2 -(1'-propynyl) estradi-1,3,5 (10) -triene-3,17β-diol, 2-azidoestradi 1,3,5 (10) -triene-3,17β-diol, 2-ethoxyestradi -1,3,5 (10) -triene 3,17β-diol, estra-1,3,5 (10) -trien-3-ol; 16-substituted 2-methoxyestradiol derivative: 2-methoxy-16α-methylestradi -1,3,5 (10) -triene-3,17β-diol 2-methoxy-16β-methylestradi-1,3,5 (10) -triene-3,17β-diol, 2-methoxy-16-ethylestradi-1,3,5, (10) -triene-3, 17β-diol, 16α- (hydroxymethyl) estradia-1,3,5 (10) -triene-3,17β-diol, 16β- (hydroxymethyl) estradia-1,3,5 (10) -triene-3, 17β-diol, 2-methoxy-16α-propaneestradi-1,3,5 (10) -triene-3,17β-diol, 2-methoxy-16β-propaneestradi-1,3,5 (10) -triene- 3,17β-diol, 2-methoxy-16β-butaneestradi-1,3,5 (10) -triene-3,17β-diol, 2-methoxy-16α-butaneestradi-1,3,5 (10)- Triene-3,17β-diol, 2-methoxy-16β-iso-butaneestradi-1,3,5 (10) -triene-3,17β-diol, 2-methoxy-16α- (N, N-dimethylaminomethyl ) Estra-1,3,5 (10) -triene-3,17β-diol, 2-methoxy-16α-ethylestradia-1,3,5 (10) -triene-3,17β-dioe Dehydrogenated / substituted 2-methoxyestradiol derivatives: 2-methoxy-estradi-1,3,5 (10) 9 (11) -tetraene-3,17β-diol; other 2-methoxyestradiol derivatives: 2-ethoxy- 19-norpregnane-1,3,5 (10) 17 (20) -tetraen-3-ol, 2- (1-propynyl) -19-norpregnane-1,3,5 (10) 17 (20) -tetraene- 3-ol, 2-formyl-19-norpregnane-1,3,5 (10) 17 (20) -tetraen-3-ol, 2-formamide-19-norpregnane-1,3,5 (10) 17 (20 ) -Tetraen-3-ol, 2-methenehydroxy-19-norpregnane-1,3,5 (10) 17 (20) -tetraen-3-ol, 2-ethyl-19-norpregnane-1,3,5 (10) 17 (20) -Tetraen-3-ol, 2-methyl-19-norpregnane-1,3,5 (10) 17 (20) -tetraen-3-ol, 2- (1-propenyl) -19 -Norpregnane-1,3,5 (10) 17 (20) -tetraen-3-ol, 2-ethoxy-17 (20) -methyleneestradi-1,3,5 (10) -trien-3-ol 2- (1-propynyl) -17 (20) -methyleneestradi-1,3,5 (10) -trien-3-ol, 2-formyl-17 (20) -methyleneestradi-1,3,5 (10) -trien-3-ol, 2-formamide-17 (20) -methyleneestradi-1,3,5 (10) -trien-3-ol, 2-methylenehydroxy-17 (20) -methyleneestradi 1,3,5 (10) -trien-3-ol, 2-ethyl-17 (20) -methyleneestradi-1,3,5 (10) -trien-3-ol, 2-methyl-17 (20) -Methylene estra-1,3,5 (10) -trien-3-ol, 2- (1-propenyl) -17 (20) -methylene estra-1,3,5 (10) -trien-3-ol, 2-Ethoxyestradi-1,3,5 (10) -trien-3-ol, 2- (1-propynyl) estradi-1,3,5 (10) -trien-3-ol, 2-formylestradi-1 , 3,5 (10) -trien-3-ol, 2-formamide estra-1,3,5 (10) -trien-3-ol, 2- (methylenehydroxy) estradia-1,3,5 (10) -Trien-3-ol, 2-ethylestradi-1 , 3,5 (10) -trien-3-ol, 2-methylestradi-1,3,5 (10) -trien-3-ol, 2- (1-propenyl) estradi-1,3,5 (10 ) -Trien-3-ol, 2-ethoxy-17β-methylestradi-1,3,5 (10) -trien-3-ol, 2- (1-propynyl) -17β-methylestradi-1,3,5 (10) -Trien-3-ol, 2-formyl-17β-methylestradi-1,3,5 (10) -trien-3-ol, 2-formamido-17β-methylestradi-1,3,5 (10 ) -Trien-3-ol, 2-methylenehydroxy-17β-methylestradi-1,3,5 (10) -trien-3-ol, 2-ethyl-17β-methylestradi-1,3,5 (10) -Trien-3-ol, 2-methyl-17β-methylestradi-1,3,5 (10) -trien-3-ol, 2- (1-propenyl) -17β-methylestradi-1,3,5 ( 10) -trien-3-ol, 2-ethoxy-17β-ethylestradi-1,3,5 (10) -trien-3-ol, 2- (1-propynyl) -17β-ethylestradi-1,3, 5 (10) -trien-3-ol, 2-e Mil-17β-ethylestradi-1,3,5 (10) -trien-3-ol, 2-formamido-17β-ethylestradi-1,3,5 (10) -trien-3-ol, 2-methylenehydroxy -17β-ethyl estra-1,3,5 (10) -trien-3-ol, 2-ethyl-17β-ethyl estra-1,3,5 (10) -trien-3-ol, 2-methyl-17β -Ethyl estra-1,3,5 (10) -trien-3-ol, 2- (1-propenyl) -17β-ethyl estra-1,3,5 (10) -trien-3-ol, 2-ethoxy -17 (20) -propylene estra-1,3,5 (10) -trien-3-ol, 2- (1-propynyl) -17 (20) -propylene estra-1,3,5 (10) -triene -3-ol, 2-formyl-17 (20) -propylene estra-1,3,5 (10) -trien-3-ol, 2-formamide-17 (20) -propylene estra-1,3,5 ( 10) -Trien-3-ol, 2-methylenehydroxy-17 (20) -propylene estra-1,3,5 (10) -trien-3-ol, 2-ethyl-17 (20) -propylene estra- 1,3,5 (10) -trien-3-ol, 2-methyl-17 (20) -propylene estra-1,3,5 (10) -trien-3-ol, 2- (1-propenyl)- 17 (20) -propylene estra-1,3,5 (10) -trien-3-ol, 2-methoxy-17β-methylenehydroxyestradi-1,3,5 (10) -trien-3-ol, 2- Methoxy-17β- (carboxylic acid) -estradi-1,3,5 (10) -trien-3-ol, colchicine, combretastatin A-4, diethylstilbestrol, 2-bromoestradiol, 2-methoxyestrone 2-hydroxyestradiol, 4-hydroxyestradiol, 17-ethynylestradiol, 2-fluoroestradiol, estradiol, estrone, 2-methoxy-17-ethynylestradiol, estriol, 2-methoxyestriol, estradiol-3-O-methyl Ether, 2-methoxyestradiol-3-O- Methyl ether, 4-methoxyestradiol, 4-methoxestradiol-3-O-methylether, podophyllotoxin, dihydrocombretastatin A-4, 3-benzyl-2-methoxyestradiol, 3-benzyl-2-methoxy Estrone, 16α-alkyl-3-benzyl-2-methoxyestrone, 16β-alkyl-3-benzyl-2-methoxyestrone, 16-alkyl-16-carbomethoxy-3-benzyl-2-methoxyestrone, 16-methane- Dimethyleneamine-3-benzyl-2-methoxyestrone, 16-carbomethoxy-3-benzyl-2-methoxyestrone, 16-alkyl-3-benzyl-2-methoxyestradi-17β-diol, 16-methanol-3- Benzyl-2-methoxyestradiol, 16-alkyl-3-benzyl-2-methoxyestradiol, 16β-methyl-2-methoxyestradiol, 16α-methyl-2-methoxyest Toradiol, 16-ethyl-2-methoxyestradiol, 16α-n-propyl-2-methoxyestradiol, 16β-n-propyl-2-methoxyestradiol, 16β-n-butyl-2-methoxyestradiol, 16β-isobutyl-2 -Methoxyestradiol, 16β-methyl (dimethylamine) -2-methoxyestradiol, 16β-methanol-2-methoxyestradiol, 2-methoxy-17-deoxyestrone, 17-ethyl-2-methoxyestrone, 17-methyl-2- Methoxyestrone, 2-N, N-dimethylamino-17-deoxyestrone, 2-azido-estradiol, and 16α-methanol-2-methoxyestradiol (e.g., U.S. Patent 6,528,676 and U.S. Patent Application 2002/0147183 and No. 2002/0082433).

  In certain embodiments, 2-methoxyestradiol or analogs and compositions useful in the methods described herein are provided to the user for the purposes described herein in the methods described herein. You may supply with the kit with the instruction | indication which shows the instruction | indication for using. The instructions for use may be printed on a container containing the composition or on a separate sheet included with the composition. In particular, the instructions may provide instructions for the user to use the composition, for example, and the purpose of such use is, for example, premature birth complications such as IVH, NEC, ROP and hemangioma formation. May indicate that it is to inhibit or otherwise prevent symptoms of symptoms or symptoms associated therewith. The instructions may be for use in individuals who are likely to have (or are predisposed to) premature complications and / or individuals who have already been diagnosed with preterm complications.

C. Formulations The compositions described herein may be administered by a variety of means depending on their intended use, as is well known in the art. For example, when the compositions are administered orally, they may be formulated as tablets, capsules, granules, powders or syrups. A liquid formulation of the composition may be formulated as a breast milk substitute or “infant formula”. In certain embodiments, 2-methoxyestradiol or an analog thereof is formulated for oral administration as a small tablet or aqueous solution or suspension.

  In certain embodiments, a liquid composition of the formulation, eg, an aqueous solution or suspension, is administered enterally, eg, by gavage from the stomach tube. Forced nutritional administration is routine and well known to those skilled in the art. Such suspensions and solutions may optionally be formulated with sweetening media, such as syrups or elixirs. Suspensions or solutions are formulated at specific concentrations, and each unit dose, for example, 1 milliliter of solution, 1 small spoon, 1 large spoon, etc. is, for example, 0.25 mg to 1250 mg of 2-methoxyestradiol or analogs (or combinations thereof) For example, a metering / dosing device such as a tube, scissors or pipette. In certain embodiments, the suspension or solution is administered enterally by gavage.

  Alternatively, the preparation may be administered parenterally by injection (intravenous, intramuscular, transdermal patch, aerosol or subcutaneous), infusion preparation or suppository. These preparations may be formulated by conventional means, and if desired, the composition can be prepared by mixing the excipient with an excipient, binder, disintegrant, lubricant, corrective, solubilizer, suspending agent, emulsifier. Or you may mix with arbitrary normal additives, such as a coating agent.

  In the formulations described herein, wetting agents, emulsifiers, lubricants such as sodium lauryl sulfate and magnesium stearate, and colorants, release agents, coating agents, sweetening, flavoring and flavoring agents, preservatives, The antioxidant may be contained in the formulated drug.

  The compositions of the present invention may be suitable for oral, nasal, topical (including buccal and sublingual), rectal, vaginal, aerosol and / or parenteral administration. The formulations may conveniently be provided in unit dosage form and may be prepared by any method well known in the pharmaceutical art. The amount of composition that can be mixed with a carrier material to produce a single dose will vary depending upon the subject being treated, the severity of premature labor complications, the state of medication, and the particular mode of administration.

  The methods of preparing these formulations include the step of mixing the compositions described herein with a carrier and optionally one or more accessory ingredients. In general, the formulations are prepared by uniformly and intimately mixing the drug with liquid carriers or finely divided solid carriers or both, and then if necessary shaping the product.

  Formulations suitable for oral administration include capsules, cachets, pills, tablets, lozenges (flavoring bases, usually sucrose and acacia) each containing a predetermined amount of the composition of the invention as an active ingredient. Or with tragacanth), in the form of powders, granules, or as a solution or suspension in an aqueous or non-aqueous liquid, or as an oil-in-water or water-in-oil liquid emulsion, or as an elixir or syrup. Or as a pasty tablet (using an inert base such as gelatin and glycerin or sucrose and acacia). The compositions described herein may be administered as a bolus, electuary or paste.

  In solid dosage forms for oral administration (capsules, tablets, pills, dragees, powders, granules, etc.), the composition of the present invention may be sodium citrate or dicalcium phosphate, and / or any of the following: Mixed with one or more pharmaceutically acceptable carriers: (1) a filler or bulking agent such as starch, lactose, sucrose, glucose, mannitol, and / or silicic acid; (2) for example, carboxymethylcellulose, Binding agents such as alginate, gelatin, polyvinylpyrrolidone, sucrose, and / or acacia; (3) humectants such as glycerol; (4) agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and carbonic acid Disintegrants such as sodium; (5) dissolution retardants such as paraffin; (6) quaternary ammonium compounds, etc. Absorption retardants such as absorption enhancers or crystallographic ingredients; (7) wetting agents such as acetyl alcohol and glycerol monostearate; (8) absorbents such as kaolin and bentonite clay; (9) talc, calcium stearate, Lubricants such as magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, and mixtures thereof; and (10) colorants. In the case of capsules, tablets and pills, the composition may contain a buffer. Similar types of solid compositions may be used as fillers in gelatin soft and hard capsules using lactose or lactose and excipients such as high molecular weight polyethylene glycols.

  A tablet may be made by compression or molding, optionally with one or more accessory ingredients. Compressed tablets use binders (e.g. gelatin or hydroxypropyl methylcellulose), lubricants, inert diluents, preservatives, disintegrants (e.g. sodium starch glycolate or crosslinked sodium carboxymethylcellulose), surface active or dispersing agents. May be prepared. Molded tablets may be prepared by molding in a suitable machine a mixture of the composition of the invention moistened with an inert liquid diluent. Other solid dosage forms such as tablets and dragees, capsules, pills and granules may optionally be scored, or enteric coatings and other coatings well known in the pharmaceutical-pharmaceutical art It may be prepared with a coating and shell.

  Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs. In addition to the composition of the present invention, the liquid dosage form can be, for example, water or ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, oil (especially Cottonseed oil, peanut oil, corn oil, germ oil, olive oil, castor oil and sesame oil), fatty acid esters of glycerol, tetrahydrofuryl alcohol, polyethylene glycol and sorbitan, and mixtures thereof, such as solubilizers and emulsifiers, It may contain inert diluents commonly used in the art.

  Suspending agents include, in addition to the compositions of the present invention, for example, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar and tragacanth, and mixtures thereof. A suspending agent may be included.

  Formulations for rectal or vaginal administration may be provided as suppositories, which comprise one or more suitable non-ionic compositions including, for example, cocoa butter, polyethylene glycol, suppository waxes or salicylates. It may be prepared by mixing with an irritating excipient or carrier, and it is a solid at room temperature but is a liquid at body temperature and therefore will melt in the body cavity to release the active agent. Formulations suitable for vaginal administration also include pessaries, tampons, creams, gels, pastes, foams or spray formulations, including carriers known to be suitable in the art.

  Dosage forms for transdermal administration of the compositions of the present invention include powders, sprays, ointments, pastes, creams, lotions, gels, solutions, patches and inhalants. The active component may be mixed under sterile conditions with a pharmaceutically acceptable carrier, and any preservatives, buffers, or propellants that may be required.

  Ointments, pastes, creams and gels, in addition to the compositions of the present invention, include animal and vegetable fats, oils, waxes, paraffins, starches, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonite, silicic acid, talc and It may contain excipients such as zinc oxide or mixtures thereof.

  Powders and sprays may contain excipients such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicate and polyamide powder, or mixtures of such substances, in addition to the compositions of the present invention. Good. The propellant may further contain customary propellants such as chlorofluorohydrocarbons and volatile unsubstituted hydrocarbons such as butane and propane.

  Alternatively, the compositions and compounds described herein may be administered by aerosol. This is accomplished by preparing an aqueous aerosol, liposomal formulation or solid particles containing the compound. Non-aqueous (eg, fluorocarbon propellant) suspensions can also be used. Sonic nebulizers may be used to minimize exposure of the agent to shear forces that can cause degradation of the compounds contained in the compositions of the present invention.

  Ordinarily, an aqueous aerosol is prepared by formulating an aqueous solution or suspension of the composition of the invention with conventional pharmaceutically acceptable carriers and stabilizers. Carriers and stabilizers vary depending on the requirements of the particular composition of the invention, but typically are non-ionic surfactants (tween, pluronic or polyethylene glycol), harmless proteins such as serum albumin, sorbitan Amino acids such as esters, oleic acid, lecithin, glycine, buffers, salts, sugars or sugar alcohols are included. Aerosols are generally prepared from isotonic solutions.

  Pharmaceutical compositions that may be suitable for parenteral administration include compositions of the invention as one or more pharmaceutically acceptable sterile isotonic aqueous or non-aqueous solutions, dispersions, suspensions or milks. In suspension, or in combination with sterile powders that can be reconstituted into sterile injectable solutions or dispersions just prior to use, which contain antioxidants, buffers, bacteriostats, formulations with the intended recipient's blood It may contain isotonic solutes or suspending or thickening agents.

  Examples of suitable aqueous and non-aqueous carriers that can be used in the pharmaceutical compositions described herein include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol, and the like), and suitable mixtures thereof, olive oil, and the like. Vegetable oils, and injectable organic esters such as ethyl oleate. The proper fluidity may be maintained, for example, by the use of a coating material such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants.

  In certain embodiments, the compounds of the invention are formulated as tablets, pills, capsules or other suitable ingestible formulations (hereinafter collectively “tablets”) to provide a therapeutic dose in 10 tablets or less. May be. In another example, the therapeutic dose is provided in 50, 40, 30, 20, 15, 10, 5 or 3 tablets.

  A tablet form of 2-methoxyestradiol or an analogue thereof is an effective amount (at least 50%) when the amount of 2-methoxyestradiol or an analogue thereof (or combination thereof) provided in 20 tablets is taken together ( ED50), eg, at least 50% of individuals may be formulated to provide a dose that has shown a counting effect of reducing at least one symptom associated with preterm complications. In a further embodiment, the total amount of 2-methoxyestradiol or an analogue thereof (or combination thereof) provided in 10, 5, 2 or 1 tablet provides at least an ED50 dose to the patient (human or non-human mammal). The tablet is formulated. In other embodiments, the amount of 2-methoxyestradiol or an analog thereof (or combination thereof) provided in 20, 10, 5 or 2 tablets taken over 24 hours averages at least 2 ED50 concentrations. -Provide a dosing regimen that provides mean plasma levels of methoxyestradiol or analogs (or combinations thereof). In other embodiments, less than 100 times, 10 times, or 5 times the ED50 is provided. In other embodiments, a single dose tablet (1-20 tablets) provides about 0.25 mg to 1250 mg of 2-methoxyestradiol or an analog thereof (or combinations thereof). In one exemplary embodiment, a single dose tablet (1-20 tablets) provides about 0.5 to 5 mg, about 1 to about 3 mg, or about 2.5 mg of 2-methoxyestradiol or an analog thereof (or combinations thereof). To do.

  Similarly, 2-methoxyestradiol or an analog thereof can be formulated for parenteral administration, e.g., subcutaneous, intramuscular or intravenous injection, e.g., 2-methoxyestradiol or an analog thereof can be formulated as a sterile solution or It can be provided in suspension (hereinafter collectively referred to as “injection solution”). An injectable solution is an effective amount of 2-methoxyestradiol or an analog thereof (or a combination thereof) provided in a 200 cc bolus injection at least 50% effective amount, or less than 100 times ED50, or 10 or 5 of ED50 Formulate to provide less than double dose. The injectable solution is a ED50 dose of the total amount of 2-methoxyestradiol or its analog (or a combination thereof) provided in an injection 100, 50, 25, 10, 5, 2.5, or 1 cc It may be formulated to provide a dose less than 100 times, or less than 10 or 5 times the ED50. In other embodiments, the amount of 2-methoxyestradiol or an analog (or combination thereof) provided in a total amount of 100 cc, 50, 25, 5 or 2 cc injected at least twice during a 24 hour period is on average A dosage regimen that provides an average plasma level of 2-methoxyestradiol or an analog thereof (or combinations thereof) that is at least an ED50 concentration, or less than 100 times ED50, or less than 10 or 5 times ED50. In other embodiments, the single dose injection provides from about 0.25 mg to 1250 mg of 2-methoxyestradiol or an analog thereof (or combinations thereof). In one exemplary embodiment, a single dose of injection provides about 0.5 to 5 mg, about 1 to about 3 mg, or about 2.5 mg of 2-methoxyestradiol or an analog (or combination thereof).

EXAMPLES Although the invention has been generally described, it will be more readily understood by reference to the following examples, which are intended to illustrate certain aspects and embodiments of the invention. It is intended as an illustration only and is not intended to limit the invention.

Example 1: Role of 2-methoxyestradiol in complications due to preterm birth
2-methoxyestradiol (2ME2) is a natural metabolite of estrogen with little or no estrogenic activity. 2ME2 is a potent angiogenesis inhibitor that suppresses the expression of hypoxia-inducible factor 1 alpha (HIF-1α). HIF-1α is a pro-angiogenic transcription factor that stimulates transcription of a plurality of pro-angiogenic proteins including vascular endothelial growth factor (VEGF). VEGF is an endothelial mitogen and survival factor that induces intravascular permeability. VEGF is 50,000 times more potent than histamine as a vascular leak agent.

  The median 2ME2 in men is <10 pg / mL, and for women of reproductive age, 46 pg / mL in the follicular phase and 70 pg / mL in the luteal phase. During pregnancy, maternal serum 2ME2 rises dramatically, reaching a median of 674 pg / mL between 11 and 16 weeks. Serum levels show a maximum of 3768 pg / mL by 37-40 weeks of gestation, which is an almost 100-fold increase over pre-pregnancy levels. Increased 2ME2 levels during pregnancy are thought to protect against a rapid increase in HIF-1α in response to brief episodes of hypoxia in the placenta and fetus.

  Neonatal umbilical cord serum contains 2ME2 at a median of 1606 pg / mL, which is about 40% of the level in maternal blood flow at term (see FIG. 1). There are no data on 2ME2 levels in cord blood or serum of premature infants, amniotic fluid of early or term infants, or colostrum or breast milk of their mothers. Furthermore, no data has been published about when this level will return to “normal” in newborns, whether preterm or full-term. However, by extrapolating from the aforementioned data that the fetal 2ME2 level is 40% of the maternal level, we predicted that the 2ME2 level at birth of the 25 week gestational infant was approximately 750 pg / mL. ing. Furthermore, if the half-life of 2ME2 in premature infants is approximately equivalent to 10 hours of adult half-life, 2ME2 would fall to less than 25 pg / mL within 2.5 days after birth.

  We found that 2ME2 levels at birth are directly proportional to gestational age, so the earlier the infant is born, the lower the 2ME2 level at birth and the level increases with time in the uterus, and finally I think it will reach the level of full-term infants. Because preterm infants are not expected to produce large amounts of their own 2ME2, they are limited to those provided by maternal and placental sources, and their levels are expected to decline rapidly based on a half-life of 2ME2 of approximately 10 hours . Since 2ME2 is essential to protect against HIF-1α, hypoxia in infants with low 2ME2 levels may lead to a rapid increase in HIF-1α, followed by downstream events such as increased circulating VEGF.

Low levels of 2ME2 and the resulting high levels of VEGF in vivo are thought to contribute to many complications of preterm birth. This can cause intraventricular hemorrhage (IVH), necrotizing enterocolitis (NEC), retinopathy of prematurity (ROP), and hemangioma for two reasons. The first reason is the loss of the direct angiogenesis-inhibiting effect of 2ME2, and the second reason is that VEGF is elevated due to the lack of HIF-1α inhibition, resulting in vascular leakage and budding formation. In summary:
Full-term infants exposed to a normal 2ME2 surge in the third trimester:
↑ 2ME2 → ↓ HIF-1 <→ ↓ VEGF (and normal angiogenesis)
Preterm infants born before the 2ME2 surge in the third trimester:
↓ 2ME2 → ↑ HIF-1 <→ ↑ VEGF → ↑ Vessel leakage (IVH, NEC) and ↑ Angiosprouting (ROP, hemangioma)

  Amniotic fluid, umbilical cord blood, or maternal serum may have protein biomarkers that can predict whether an infant is at increased risk of developing IVH, NEC, and ROP before showing clinical signs . Such a series of markers may help to find out which patients are at higher risk of developing these complications. We hypothesize that the balance of pro-angiogenic and inhibitory factors is relatively inclined to excessive angiogenesis in preterm infants compared to the control group matched by gestational age in utero. . Excessive angiogenesis is thought to be due to a relative deficiency of the potent angiogenesis inhibitor 2-methoxyestradiol (2ME2) in preterm infants, which includes IVH, retinal neovascularization and ROP, NEC As well as an increased risk of life-threatening hemangiomas.

  The present inventors have shown that a steady increase in the level of 2-methoxyestradiol in the blood of pregnant women occurs in parallel in the blood of the fetus, and the hypoxia-inducing factor in response to a brief onset of hypoxia in the placenta— The hypothesis is that the fetus will be protected against a surge of 1 alpha (HIF-1 alpha).

  In order to compare various pro-angiogenic and inhibitory factors and collect outcome data on IVH, NEC, ROP and hemangioma as described in the examples below, we have premature infants and their mothers and Blood, amniotic fluid and breast milk are collected from a healthy full-term control group. A unified molecular basis for the increased incidence of morbidity associated with premature infants will be provided, leading to prophylactic treatment.

  Identifying which preterm infants will develop IVH, NEC, or ROP prior to the onset of clinical symptoms at birth will allow for treatment tailored to these infants. Replacement therapy with the correct angiogenic cocktail given orally or parenterally appears to be sufficient to prevent these complications (Figure 2). At present, 2ME2 is being tested in clinical trials (Phases 2 and 3) for advanced prostate cancer and no side effects have been reported. Of note, this inhibits abnormal (eg, oncological) blood vessel growth, but does not appear to affect normal blood vessel growth or activity. This will ultimately provide a theoretical opportunity to supplement 2ME2 if it is found that premature infants are actually relatively deficient in this angiogenesis inhibitor.

  In particular, the following questions will be addressed during the study.

  1. Is there a link between early labor and low 2ME2 levels? By comparing the 2ME2 level of pregnant women in early labor with a control group matched by gestational age that is not in early labor, you can see if there is an association between 2ME2 levels and early labor (PTL), A causal question arises whether low 2ME2 induces preterm birth.

  2. Betamethasone, administered to mothers before preterm birth to mature lung function, also reduces the incidence of IVH for unknown reasons. Does betamethasone stimulate 2ME2 production by the placenta, a possible mechanism of reduced IVH incidence?

  3. Preterm infants that will eventually develop IVH, NEC, ROP, or hemangioma are born at birth before signs of disease using a set of maternal serum, amniotic fluid, or cord blood protein biomarkers. Can it be identified?

  4. Are preterm infants who develop IVH, NEC, ROP, or hemangioma have lower 2ME2 levels at birth compared to premature infants who are age-matched to these uncomplicated ages?

  5. What is the 2ME2 half-life for infants? Does a short half-life pose a risk of inappropriately short protection against a hypoxic outbreak?

  6. Can the rate of infant decline be predicted from the 2ME2 decline rate of premature mothers?

  7. Amniotic fluid theoretically contains 2ME2 and other pro-angiogenic and inhibitory factors, which may be a source of balanced angiogenic “cocktails”.

  8. Breast milk is thought to contain 2ME2 which can provide protection for preterm infants. What is the level of 2ME2 in breast milk, and are preterm infants breast-fed protected against IVH, NEC, ROP and hemangioma compared to preterm infants not breast-fed?

Desktop Example 2: Evaluation of 2-methoxyestradiol levels at birth (umbilical cord blood, amniotic fluid, maternal placenta), and 2ME2 half-life in infant plasma and breast milk Using gas chromatography-mass spectrometry (GC-MS) Levels of 2ME2 or analogs such as cord blood, amniotic fluid, plasma, breast milk can be measured. GC-MS is a standard test method for detecting and quantifying low molecular weight steroids (Mw = 302 g / mol) such as 2-methoxyestradiol.

  We have improved our experimental procedures, including examining the broad range and dynamics of 2ME2 levels in pregnant women and newborns to fine-tune how many samples are needed from each infant Start with a pilot test. Therefore, the present inventors have found that 5 mothers (Example 2a) of full-term infants born by caesarean section (37-42 weeks of gestation), 5 mothers and their full-term infants requiring hospitalization of NICU (Example) 2b) and about 270 samples of 2ME2 from 10 mothers and their very premature babies (pregnancy <28 weeks) (Example 2c) are measured. These samples, as well as all other samples under test, are in the 2-10 mL volume.

  Because blood samples cannot be collected from normal term infants (since term infants do not need any blood for normal analysis), we have not obtained umbilical cords from normal term infants born after cesarean section. Collect only blood, maternal serum, amniotic fluid, and maternal colostrum. The 2ME2 half-life is measured in term infants (37-40 weeks of gestation), which requires hospitalization with NICU and multiple blood analyzes over time.

  Data from term infants will serve as controls to determine the extent of 2ME2 levels in newborns and breast milk and to assess the rate of decline in 2ME2 levels. Analyze data from preterm infants to see if 2ME2 in preterm infants follow the same dynamics as full-term infants. These data are based on the time when 2ME2 levels can no longer be detected in preterm and full-term infants and improve our experimental assay to determine how many samples need to be taken from the remainder of preterm infants. I think it will help to investigate.

Example 2A: Mother of full-term infant by pilot test, caesarean section
Patient Registration / Exclusion Criteria: 37 0 / 7-42 complete weeks of pregnancy at birth, cesarean delivery, healthy after delivery, preferably breastfeeding expected
Number of patients: 5 mothers
Samples to collect: All samples = 50

Example 2B: Pilot trial, mother and the sick full-term infant admitted to NICU
Patient Registration / Exclusion Criteria: 37 0 / 7-42 complete weeks of pregnancy at birth, hospitalization at NICU for any reason is expected to allow more than 4 days of hospitalization, breastfeeding schedule
Number of patients: 5 mothers and 5 infants
Samples to collect: All samples = 60

DOL＝生後日数
＾ 常に臨床的に指示された採血と連係して行う。
* 10日または退院のいずれか早い方。

DOL = days after birth ^ Always in conjunction with clinically directed blood collection.
* 10 days or discharge date, whichever comes first.

Example 2C : Pilot study, mother and its preterm infant
Patient registration / exclusion criteria: Pregnancies under 28 weeks of pregnancy (eg, 28 6/7 weeks), preferential breastfeeding due to cesarean delivery
Number of patients: 10 mothers and 10 infants
Samples to collect: All samples = 160

DOL＝生後日数
＾ 常に臨床的に指示された採血と連係して行う。
* 10日または退院時のいずれか早い方。
血液量の詳細：
目標はいつでも可能なときに1ccを送ること
要求：8週間毎に3cc/kg
0.5kgの乳児では、最大1.5cc＝4×0.37cc
1kgの乳児では、最大3cc＝4×0.75ccの試料
1.5kgの乳児では、最大4.5cc＝4×1.1ccの試料
2kgの乳児では、最大6cc＝4×1.5ccの試料

DOL = days after birth ^ Always in conjunction with clinically directed blood collection.
* 10 days or discharge time, whichever comes first
Blood volume details:
The goal is to send 1cc whenever possible: 3cc / kg every 8 weeks
For a 0.5 kg infant, maximum 1.5 cc = 4 x 0.37 cc
For a 1kg infant, a maximum of 3cc = 4 x 0.75cc sample
For a 1.5 kg infant, a maximum of 4.5 cc = 4 x 1.1 cc sample
For 2 kg infants, maximum 6 cc = 4 x 1.5 cc sample

  Samples are preferably centrifuged within 30 minutes after blood collection in a Li-heparin tube (except for breast milk) and the plasma phase is frozen at -70 ° C.

Desktop Example 3: Correlation of a series of pro-angiogenic and inhibitory factors measured in cord blood and outcome data for IVH, NEC, ROP and hemangioma formation Using multiple antibody platforms and ELISAs, the inventors Examines the relative abundance of different growth factors to assess the net “angiogenic balance” by gestational age and correlate it with IVH, NEC, ROP and infantile hemangioma formation. For this purpose, amniotic fluid and umbilical cord blood samples are collected or stored samples are used if available. Use 2-5 mL samples from infants less than 32 weeks of gestation (eg, 32 6/7 weeks) from which cord blood can be collected as discarded human material.

  Prevalence of IVH (any grade), NEC and ROP (any stage) is 5-10%, and umbilical cord blood from 15-20 infants developing their respective complications Based on the goal of collection, we estimate that approximately 300 cord blood samples need to be collected.

  We collect umbilical cord blood as waste human material from the mother / infant pair enrolled in this part of the study. Based on our hypothesis, these four complications arise mainly from the pro-angiogenic environment. Umbilical cord blood is analyzed for the following factors and correlated with outcome data on IVH, NEC, ROP and hemangioma formation.

1. 2ME2 plasma levels to determine if these angiogenesis inhibitor levels are low at birth in infants with these four complications

2. Plasma levels of multiple angiogenesis and angiogenesis-inhibiting markers We found that infants who develop IVH, NEC, ROP, and hemangioma were more likely to have other phenotypes at birth compared to controls matched at gestational age. We want to investigate whether the level of known angiogenesis inhibitors (such as thrombospondin) is low or the level of angiogenic growth factors (such as VEGF, bFGF, PlGF) is high. We use a multiplex monoclonal antibody microarray system that allows simultaneous measurements of tens to hundreds of biomarkers in small amounts.

3. Biomarker discovery If angiogenesis / anti-angiogenesis profiling finds that biomarkers that can accurately predict patient outcomes are not found, we can use two-dimensional gel electrophoresis and the DIGE system. Identify novel proteins that can serve as biomarkers for IVH, NEC, ROP or hemangioma.

Desktop Example 4: Discovery and quantification of protein biomarkers in cord blood Detection of a series of biomarkers provides greater sensitivity and specificity for diagnosis and prognosis than single markers. The present inventors investigate whether changes in protein abundance in umbilical cord plasma can be an index to predict IVH, NEC, ROP and infantile hemangioma formation before clinical manifestations in premature infants. We use the DIGE system (GE) to discover new protein biomarkers.

Equivalents While specific aspects of the invention have been discussed, the foregoing specification is illustrative and not restrictive. Upon careful reading of the specification, many variations of the present invention will become apparent to those skilled in the art. The appended claims are not intended to claim all such embodiments and variations, and the full scope of the present invention includes the claims and their equivalents, and the specification and its equivalents. Should be determined by referring to such variants.

  All publications and patents mentioned in this specification, including those listed below, are all in their entirety as if each individual publication or patent was specifically and individually indicated to be incorporated by reference. Which is incorporated herein by reference. In case of conflict, the present application, including any definitions herein, will control.

Shown are maternal serum levels of 2ME2 during pregnancy and umbilical cord blood values of term infants (black letters). Extrapolated maternal and umbilical cord blood levels (red letters, parentheses) for premature infants at 25 weeks of age. Expected changes in umbilical cord blood levels of 2ME2 (blue line) corresponding to fetal serum levels in the uterus, based on half-life measurements of adult 2ME2, and predicted decrease in 2ME2 serum levels in 25-week-old preterm infants (red line) ). Note that the red numbers and lines are extrapolated. 2 shows the recovery of physiological levels of 2ME2 in very preterm infants to that seen in the serum of the control group matched by gestational age.

Claims (36)

  A method of treating or preventing complications of preterm birth, wherein a subject in need thereof is effective of 2-methoxyestradiol (2-ME), an analog thereof, or a pharmaceutically acceptable salt or prodrug thereof Administering a quantity.   The method of claim 1, further comprising first identifying a subject susceptible to preterm birth complications.   The method of claim 1, further comprising first identifying a subject suffering from preterm complications.   2. The method of claim 1, further comprising quantifying the level of 2-methoxyestradiol and / or its analog in a biological sample from the subject.   5. The method of claim 4, wherein the level of 2-methoxyestradiol and / or its analog is quantified by quantifying the level of precursor or metabolite of 2-methoxyestradiol and / or its analog.   5. The method of claim 4, further comprising comparing the level of 2-methoxyestradiol and / or its analog to the level of 2-methoxyestradiol in a control.   2. The method of claim 1, further comprising quantifying the level of precursor of 2-methoxyestradiol in the biological sample from the subject.   8. The method of claim 7, further comprising comparing the level of 2-methoxyestradiol precursor to the level of 2-methoxyestradiol precursor in a control.   2. The method of claim 1, further comprising quantifying the level of 2-methoxyestradiol and / or its metabolite in a biological sample from the subject.   10. The method of claim 9, further comprising comparing the level of a 2-methoxyestradiol and / or analog metabolite to a level of a 2-methoxyestradiol metabolite in a control.   5. The method according to claim 4, wherein the biological sample is urine, blood, or plasma.   5. The method of claim 4, wherein the control is a pre-quantified standard from at least one subject who does not suffer from preterm complications.   If the level of 2-methoxyestradiol in the subject is lower than the level of 2-methoxyestradiol in the control, the level of 2-methoxyestradiol and / or its analog in the subject is approximately the same as the level of 2-methoxyestradiol in the control 7. The method of claim 6, further comprising administering to the subject an amount of 2-methoxyestradiol or an analog thereof effective to increase the level.   7. The method of claim 6, comprising the step of periodically quantifying the level of 2-methoxyestradiol and / or its analog in the subject relative to a control.   15. The method of claim 14, wherein the periodicity is about once every 1, 2, 3, 4, 5, 6, 7, 10, 14, 28, or 35 days.   The level of 2-methoxyestradiol or an analog administered to a subject is maintained at a level that is approximately the same as the level of 2-methoxyestradiol in a control in the subject. 15. The method of claim 14, further comprising the step of periodically adjusting to:   2. The method of claim 1, wherein the subject is a human.   2. The method of claim 1, wherein about 0.1, 0.3, 1, 2.5, or 5 mg of 2-methoxyestradiol or an analog thereof is administered to the subject.   2. The method of claim 1, wherein 2-methoxyestradiol or an analog thereof is administered orally, intranasally, parenterally or transdermally.   2. The method according to claim 1, wherein 2-methoxyestradiol or an analog thereof is administered enterally by gavage.   2. The method of claim 1, further comprising monitoring the subject for one or more symptoms of preterm complications.   2. The method of claim 1, wherein the preterm complication is selected from the group consisting of intraventricular hemorrhage, necrotizing enterocolitis, retinopathy of prematurity, and hemangioma.   2. The method of claim 1, further comprising administering to the subject at least one second therapeutic agent that is not 2-methoxyestradiol, an analog thereof, or a pharmaceutically acceptable salt or prodrug thereof.   24. The method of claim 23, wherein the second therapeutic agent is administered to the subject concurrently with 2-methoxyestradiol or an analog thereof. 2. The method of claim 1, wherein the 2-methoxyestradiol analog is a compound of formula I:

In the formula, for each occurrence independently:
R is H, alkyl, aryl, aralkyl, or carbonyl;
R1 and R2 are -OR, -SR, or -N (R) 2;
R3 is H, halide, alkyl, aryl, aralkyl, or carbonyl;
R4, R5, and R6 are H, halide, hydroxy, alkoxy, amino, thiol, alkyl, aryl, aralkyl, or carbonyl;
m is an integer from 1 to 5; and
n and p are integers from 1 to 6.   2. The method of claim 1, wherein the 2-methoxyestradiol analog is colchicine or combretastatin A-4. A method of treating or preventing preterm complications comprising the following steps:
a) quantifying the level of 2-methoxyestradiol in a biological sample from a preterm subject and comparing the level to the level of 2-methoxyestradiol in a control, wherein the control is at least one non-early subject A stage that is a pre-quantified standard from an early-birth subject having the same gestational age as the early-birth subject that does not have complications from or from the pre-birth;
b) an amount of 2-methoxyestradiol or an amount thereof sufficient to raise the level of 2-methoxyestradiol and / or its analog in the biological sample of the subject to a level approximately equal to the level of 2-methoxyestradiol in the control Administering an analog to said preterm subject;
c) periodically repeating steps a) and b) thereby maintaining the level of 2-methoxyestradiol in the subject at about the same level as the level of 2-methoxyestradiol in the control.   28. The method of claim 27, wherein the biological sample is urine, blood, or plasma.   28. The method of claim 27, further comprising the step of first identifying a subject susceptible to or suffering from preterm birth complications.   Diagnose or predict the onset of preterm complications in a subject, including quantifying the level of 2-methoxyestradiol in a biological sample from the subject and comparing the level to the level of 2-methoxyestradiol in a control The method wherein the control is pre-derived from a subject who has no complications of at least one premature birth and is about the same gestational age as the subject or from a subject who is at least one full-term infant. A method that is a quantified standard and that a low level of 2-methoxyestradiol in the subject sample relative to a control indicates that the subject is susceptible to or suffers from preterm complications.   The severity of preterm complications is determined based on the degree of difference in subjects' levels of 2-methoxyestradiol compared to controls, with greater deviation from controls indicating that preterm complications are more severe 32. The method of claim 30, wherein:   32. The method of claim 31, wherein the biological sample is urine, blood, or plasma.   35. The method of claim 32, wherein the subject is a human.   Quantifying the level of 2-methoxyestradiol metabolite in a biological sample from the subject and comparing the level to the level of 2-methoxyestradiol metabolite in a control to develop preterm complications in the subject A method of diagnosing or predicting, wherein the control is from a subject of about the same gestational age as the subject who has no at least one preterm complication or is at least one full-term infant A pre-quantitative standard from the body, with a low level of 2-methoxyestradiol metabolite in the subject sample compared to the control, the subject is susceptible to or suffers from preterm complications Show you how.   Quantifying the level of 2-methoxyestradiol precursor in a biological sample from the subject and comparing the level to the level of 2-methoxyestradiol precursor in a control to develop preterm complications in the subject A method of diagnosing or predicting, wherein the control is from a subject of about the same gestational age as the subject who has no at least one preterm complication or is at least one full-term infant A change in the level of 2-methoxyestradiol precursor in the subject sample relative to the control, which is a pre-quantitative standard from the body, the subject is likely to suffer from or suffers from preterm complications Show you how.   36. The method of claim 35, wherein a high level of 2-methoxyestradiol precursor in a subject sample relative to a control indicates that the subject is susceptible to or suffers from preterm complications.

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