EP0931263A1 - Screeningsferfahren für nebenwirkungen von antikonzeptionsmitteln oder östrogen und/oder progesteron ersatz- oder zusatzmitteln - Google Patents

Screeningsferfahren für nebenwirkungen von antikonzeptionsmitteln oder östrogen und/oder progesteron ersatz- oder zusatzmitteln

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Publication number
EP0931263A1
EP0931263A1 EP96930447A EP96930447A EP0931263A1 EP 0931263 A1 EP0931263 A1 EP 0931263A1 EP 96930447 A EP96930447 A EP 96930447A EP 96930447 A EP96930447 A EP 96930447A EP 0931263 A1 EP0931263 A1 EP 0931263A1
Authority
EP
European Patent Office
Prior art keywords
acute phase
compound
composition
increase
sex steroid
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Withdrawn
Application number
EP96930447A
Other languages
English (en)
French (fr)
Inventor
Cornelis Kluft
Josephus Jan Emeis
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Nederlandse Organisatie voor Toegepast Natuurwetenschappelijk Onderzoek TNO
Original Assignee
Nederlandse Organisatie voor Toegepast Natuurwetenschappelijk Onderzoek TNO
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Nederlandse Organisatie voor Toegepast Natuurwetenschappelijk Onderzoek TNO filed Critical Nederlandse Organisatie voor Toegepast Natuurwetenschappelijk Onderzoek TNO
Publication of EP0931263A1 publication Critical patent/EP0931263A1/de
Withdrawn legal-status Critical Current

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Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/74—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving hormones or other non-cytokine intercellular protein regulatory factors such as growth factors, including receptors to hormones and growth factors
    • G01N33/743—Steroid hormones

Definitions

  • Preparations with sex steroids have been developed for birth control, supplementation e.g. when women enter the menopause, replacement when women have had hysterectomies. Alternatively they are used for sex- change processes and growth inhibition for tall boys and girls.
  • the preparations used in the field of anticonception have in recent history been adapted with regard to both dosage and type of compounds. The reason was the recognition that besides the targeted action of the sex steroids in anticonception, additional effects occurred in the first generation pills and increased occurrence of cardiovascular disease in these groups was documented in epidemiological surveys .
  • the dose of the estrogenic component was identified as an important determinant and was reduced gradually with time. Up till recently, epidemiology supported these adaptations of the third generation in showing reductions in cardiovascular side effects. Newer formulations with lower dosages are being evaluated for future use. Up till recently, epidemiology supported these adaptations in showing reductions in cardiovascular side effects.
  • progestagen-only pills were not considered extensively because current evidence suggested that changes are small even by sensitive indicators of lipid and carbohydrate metabolism and components of hemostasis. Furthermore there are insufficient epidemiologic data to determine whether the progestagen-only pills influence the risk of cardiovascular diseases.” They further state that the progestagen component is mainly responsible for the effect of OCs (oral contraceptives) on carbohydrate metabolism and that the effects on hemostasis are conceivably estrogen mediated because they have not been demonstrated in progestagen-only preparations.
  • OCs oral contraceptives
  • Oral contraceptives most often contain both an estrogenic and a prostagenic compound.
  • the mechanisms involved in metabolic changes to date were considered to reflect estrogenic metabolic effects of the estrogen component and, depending upon the compound, intrinsic estrogenic, androgenic or anti-estrogenic properties of the progestagenic component. Firstly with this in mind the dosage of estrogen was lowered.
  • a set of modern, new prostagenic compounds including desogestrel, gestodene and norgestimate were in recent years introduced and they represent the so-called third generation Ocs.
  • the newer progestins were designed to minimize the adverse effects (e.g., acne, hirsuitism, nausea, carbohydrate and lipid metabolism changes) observed with older Ocs, while maintaining efficacy and good menstrual cycle control.
  • Desogestrel, norgestimate, and gestodene have minimal amounts of androgenicity and antiestrogenic potential as described in Ann-Pharmacother. 1995 Jul-Aug; 29(7 ⁇ 8): 736-42. This article reviews and compares the newer progestins desogestrel, norgestimate, and gestodene with regard to chemistry, pharmacokinetics, efficacy, and tolerability .
  • any of the combination OC products that contain these progestins may be prescribed for women intolerant of older agents" or to first-time users of OCs.
  • the newer progestins appear to be efficacious and offer similar cycle control, improved safety and tolerability profiles, and comparable price with the older agents.
  • SHBG sex-hormone-binding globulin
  • CBG cortisol-binding globulin
  • DHEAS dihydroepiandrosterone sulfate
  • the DHEAS concentration decreased less with the 20 micrograms EE + desogestrel formulation compared with either 30 micrograms EE + desogesterel or norgestimate-containing formulations (20% vs. 5%) .
  • Concentrations of SHBG and CBG increased significantly in all four groups (average 263 + / ⁇ 119% and +/- 26%, respectively); CBG increased less in women taking 20 micrograms EE + desogestrel (about 75%) than in the other formulations (about 100%).
  • the four modern, low-dose OCs tested had similar impacts on endogenous androgen metabolism, yielding significant decreases in testosterone, dihydrotestosterone, androstenedione, and DHEAS.
  • SHBG sex hormone binding globulin
  • CBG cortisol-binding globulin
  • DHEAS dihydroepiandrosterone sulfate
  • the subject invention is thus directed at a method for screening for negative side effects of a sex steroid compound or composition by assessing the level of a marker selected from a group of proteins or complexes of proteins hitherto never associated with a sex steroid compound or known negative side effects thereof in a subject after said subject has been dosed with said compound or composition.
  • a sex steroid composition is a composition useful for the applications described above in particular as contraceptive for reproduction and in hormone replacement or supplementing therapy.
  • a sex steroid composition comprises an oestrogenic and/or progestogenic compound or combination thereof.
  • a method according to the invention can be applied within the first one to six months of intake of the preparation in order to assess whether the subject is sensitive to the preparation.
  • APR acute phase reactants
  • the acute phase of the inflammatory response refers to a wide range of physiological changes initiated immediately after infection or physical trauma.
  • the ensuing cascade of mediators induces activation proliferation and altered behavior and changes in the biosynthetic capacities of a variety of target cells and tissues.
  • An important aspect of the acute phase response is the radically altered biosynthetic profile of the liver.
  • Hepatic acute phase reactants comprise the following proteins: complement proteins :C2. C3.
  • C4 binding protein proteinase inhibitors: ⁇ l-antitrypsin, ⁇ l-antichymotrypsin, ⁇ 2- antiplasmin, Heparin co actor II, metal binding proteins:ha toglobulin.
  • a number of other reactants also exist such as the major reactant Secretory Phospholipase A2 (sPLA2), ferritin, retinol binding protein, and coagulation related proteins i.e. tissue plasminogen activator and plasminogen.
  • sPLA2 major reactant Secretory Phospholipase A2
  • ferritin ferritin
  • retinol binding protein retinol binding protein
  • coagulation related proteins i.e. tissue plasminogen activator and plasminogen.
  • proteins have also been proposed to belong to the group of APRs but remain to be confirmed. This group comprises angitensinogen, kininogen, kininogenase , fibronectin, prothrombin, Factor VIII, heparin cofactor II.
  • Some of the hepatic reactants have also been illustrated to be produced extra hepatically e.g. SAA.
  • the acute phase reactants vary in degree in which they are or can be activated.
  • an acute phase reactant is defined as a reactant that can vary in concentration by 25% or more in the first 7 days of inflammation or following tissue damage.
  • the major acute phase reactants can be produced up to 1000 fold their basic level. Ceruloplasmin and C3 exhibit about 50% increase.
  • the response is known to be initiated and coordinated by a large number of inflammatory mediators, which include cytokines, anaphylatoxins and glucocorticoids.
  • cytokines include cytokines, anaphylatoxins and glucocorticoids.
  • Several hormones specifically regulate the transcripton of human APRs include interleukin 1 , interleukin 6, tumor necrosis factor, transforming growth factor ⁇ , interferon gamma, glucocorticoids and effector molecules comprising interleukin 2, oncostatin M, ciliary neurotrophic factor and retinoic acid.
  • interleukin 1 and tumor necrosis factor induce APR synthesis in the liver and stabilisation of mRNA for positive APRs occurs by cytokines and glucocorticoids. Mackiewicz A., Ganapathi M.K.
  • CRP Creprivation protein
  • pro-inflammatory component itself, and in increasing tissue trauma effects makes an increase in this component undesirable for persons defective or weak in control mechanisms of trauma reactions such as individuals with hereditary defects in the anticoagulation system (deficiencies in antithrombin III, protein S and Factor V Leiden).
  • an increase in CRP not only marks acute phase effects but also adds directly to the risk of venous thrombosis.
  • the increase in the vascular von Willebrand factor and in factor VIII itself are also considered undesirable in view of the previously already described association between elevated levels of these components and the risk of venous thrombosis (Br-Heart-J. 1995 Dec; 74(6): 580-3).
  • vWF seems to mediate platelet aggregation and adhesion to the vascular endothelium. It further states because the release of vWF is increased when endothelial cells are damaged, vWF has been proposed as an indicator of endothelial disturbance or dysfunction.
  • the availability of such an index of endothelial dysfunction is postulated as having clinical value, because measurement of such a marker can be a non-invasive way of assisting in diagnosis or as an indicator of disease progression.
  • the known association between vWF, thrombogenesis, and atherosclerotic vascular disease also suggests that high concentrations of vWF may be an indirect indicator of atherosclerosis and/or thrombosis.
  • APRs acute phase reactants
  • the group of APRs exhibiting more than 100 fold incremental change during an acute phase reaction forms an extremely suited group of proteins for carrying out a test according to the invention due to the high degree of sensitivity that can be achieved. This is in contrast to other proteins belonging to the group of APRs in general like ceruloplasmin, von Willebrand factor, Factor VIII or fibrinogen which exhibit much smaller increments.
  • the invention comprises a method for screening for negative side effects of a sex steroid compound or composition in a subject, by carrying out an assay on the subject or on a sample derived from the subject determining whether an increase of the compound or composition on the level of an acute phase reactant or a metabolic derivative thereof has occcurred since applying the compound or composition to the subject, said acute phase reactant being selected from the group consisting of positive APRs with the exclusion of ceruloplasmin and the coagulation/thrombosis associated factors described above.
  • a suitable group of embodiments of the above type comprises a method using an APR selected from metal binding proteins with the exclusion of ceruloplasmin. Another suitable group is that of proteinase inhibitors.
  • a preferred embodiment of the claimed invention is defined as ollows: A method for screening for negative side effects of a sex steroid compound or composition in a subject, by carrying out an assay on the subject or on a sample derived from the subject determining whether an increase of the compound or composition on the level of an acute phase reactant or a metabolic derivative thereof has occcurred since applying the compound or composition to the subject, said acute phase reactant being selected from the group consisting of APRs defined as being capable of exhibiting more than 100 fold increment within 7 days after tissue damage or inflammation, preferably more than 200 fold, whereby an increase in the level of the acute phase reactant is indicative of negative side effects.
  • the preferred markers to be used in the invention belong to the group of APRs known to be capable of exhibiting up to 1000 fold increase during an acute phase reaction Markers belonging to. Additionally preferred markers according to the invention belong to the group of APRs exhibiting a low or slightly variable basic level thus enabling good quantification of a disturbance in this level.
  • This group comprises the proteins serum amyloid A (SAA), serum amyloid P (SAP), C-reactive Protein (CRP) and Secretory Phospholipase A2 (sPLA2) as disclosed by D.M. Steel and A.S. Whitehead in Immunology Today 1994; 15: 81-88 by Kushner et al as mentioned elsewhere in this description.
  • the sample to be analysed in any of the embodiments of a method according to the invention can be a blood sample or any other sample suitable for assessing levels of plasma proteins as is common knowledge for a person skilled in the art.
  • the method according to the invention can be carried out in an in vitro system or in an in vivo system. Both animal and human test sytems can be used. CRP cannot be tested in a rat system however as it is absent in rats. Alternative APP proteins of the type described can however be used in a rat model. SAP is the rat homologue of CRP. CRP is one of the preferred markers for use in humans .
  • the detection methods to be used will depend on the marker to be analysed .
  • Numerous assays are already known for determining the amounts of such proteins.
  • the CRP can be determined using the CRP assay disclosed in the WHO Expert Committee on Biological Standardisation (37th Report. World Health Organ Tech. Rep. Ser. 1987; 760: 21-22 and New Engl . J. Med. 1994; 331: 417-424).
  • the SAA assay can be carried out as disclosed by J. Wilkins, J.R. Gallimore, G.A. Tennent et al in Clin Chem 1994; 40: 1284-1290.
  • SAP is described by R.L. Meek, S. Urieli-Shovall and E.P.
  • the screening will additionally involve determination of the level of a haemostatic factor, said haemostatic factor also belonging to the group of APRs.
  • haemostatic factor such factors are fibrinogen (Thompson et al cited above) , von Willebrand Factor (B.E. Pottinger, R.C. Read, E.M. Paleolog, P.G. Higgins and J.D. Pearson in Thromb. Res. 1 89; 53: 3 ⁇ 7-394), Tissue Plasminogen Activator (t-PA)(J.H. Jansson, B. Norberg, T.K. Nilsson in Clin.
  • Plasminogen Activator Inhibitor (PAI-I) (J.H. Jansson, B. Norberg, T.K. Nilsson in Clin. Chem 1989; 35: 1544-1545), plasminogen (I. Juhan-Vague, M.C. Alessi, P. Joly, X. Thirion, P. Vague, P.J. Declerck, A. Serradimigni and D. Collen in Arterisclerosis 1989; 9 - 362-367) and ⁇ 2 -anti ⁇ lasmin (Steel et al as cited above).
  • PAI-I Plasminogen Activator Inhibitor
  • a method for determining an increase in von Willebrand factor in combination with any of the preferred embodiments described above for the method according to the invention with the first APR falls within the preferred scope of the invention.
  • Determination of an increase of an APR from the group defined as capable of exhibiting more than 100 fold increase within the first 7 days after tissue damage or upon inflammation provides a suitable embodiment of the aforementioned screening method with 2 APRs.
  • Screening assays for haemostatic factors are described in large detail in the prior art. Specifically reference is made to the ECAT Assay Procedures, A manual of Laboratory techniques, European Concerted Action on Thrombosis and Disabilities of the Commission of the European Communities, edited by J. Jespersen, R.M. Bertina and F. Haverkate. The information regarding hamostatic factor assays of the factors described in the previous sentence is hereby incorporated by reference. Numerous alternatives exist and will be readily apparent to a person skilled in the art.
  • An alternative embodiment of the invention comprises an in vitro method for screening for negative side effects of a sex steroid compound or composition in a subject, by carrying out an assay on a sample that has been exposed to the compound or composition determining whether an increase of the compound or composition on the level of an acute phase reactant modulator or a metabolic derivative thereof has occcurred since applying the compound or composition.
  • the modulator being suitably selected from the group consisting of biological response modifiers.
  • biological modifiers comprise the modifiers mentioned earlier contributing to induction of APRs such as cytokines, interleukins etc. Specifically the enhancers of CRP are preferred. We hypothesize that estrogens appear to have a stimulatory effect on a package of acute phase proteins.
  • the negative side effects to be combatted are those of increased risk of cardiovascular disease such as thrombosis.
  • the negative side effects are those correlated to increase in one or more acute phase reactants.
  • the screening can occur using any of the methods according to the invention as disclosed above.
  • the compounds thus screened can be novel compounds or can be found amongst known compounds. Combination of the compounds can lead to a sex steroid preparation as safe as the third generation pills or better. It can also lead to alternative formulations at least as good as the known second generation pills.
  • the determining factor is that the new formulation comprising either a novel compound or a combination of novel compounds or a novel combination of known compounds exhibits less increase in an APR than a third generation pill preferably less than a second generation pill when these are compared.
  • a sex steroid preparation according to the invention will not result in an increase of an APR as can be determined using a method according to the invention.
  • Such alternative forms of providing medication are as such known for other fields of medicine and for some sex steroid preparations.
  • Examples of known intrauterine devices releasing a sex steroid preparation are the Progestasert which releases progesterone at a rate of 65 micrograms per day and the LNG-20 which releases 20 micrograms of levonorgestrel per day.
  • the Norplant system employs silastic tubing permeable to steroid molecules to provide stable circulating levels of levonorgestrel .
  • Depot-medroxyprogesterone acetate (Depo Provera) is given in the form of an injection as is norethindrone enanthate.
  • Transdermal application of oestrogen is also known.
  • the volunteers were randomized (open label) to the use of two different 0C preparations with an equal dose of ethinylestradiol (30 ug) , but different type and dose of progesteron, being either 75 ug gestodene (GTD-EE) or 150 ug desogestrel (DSG-EE).
  • C-reactive protein was measured with an enzyme-immuno-assay , using horseradish peroxidase labelled polyclonal antibodies against CRP ( Dako, Copenhagen, Denmark) as catching and tagging antibody in a manner known per se.
  • CRP standard serum (Beh ⁇ ngwerke, Marburg, Germany) was - --
  • Plasma CRP levels were measured by an enzyme immunoassay (ELISA) .
  • the microtitration plates were coated with 100 ⁇ l Rabbit-anti-human CRP (DAKO, Denmark) in coating buffer (1:8000) and incubated overnight at 4°C. The wells were washed three times with 0,1# PBS-Tween-20 .and ImM Na 2 EDTA.
  • the plasma samples diluted 1:1000 (Hamilton, MicroLab 1000) in PBS-Tween containing 3% polyethylene glycol 6000 were added in duplicate to the coated wells. The plates were incubated for two hours at room temperature, the wells were rewashed three times and 100 ⁇ l peroxidase- conjugated rabbit-anti-human CRP was added for one hour at room temperature.
  • C-reactive protein was measured with an enzyme-immuno-assay , using horseradish peroxidase labelled polyclonal antibodies against CRP ( Dako, Copenhagen, Denmark) as catching and tagging antibody.
  • CRP standard serum (Behringwerke, Marburg, Germany) was used to standardize the results (see detailed procedure according to Example I) .
  • Observations CRP-values Pretreatment 6 weeks
  • the pretreatment values in both groups are not statistical significant different.

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  • Life Sciences & Earth Sciences (AREA)
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EP96930447A 1996-09-06 1996-09-06 Screeningsferfahren für nebenwirkungen von antikonzeptionsmitteln oder östrogen und/oder progesteron ersatz- oder zusatzmitteln Withdrawn EP0931263A1 (de)

Applications Claiming Priority (1)

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PCT/NL1996/000350 WO1998010293A1 (en) 1996-09-06 1996-09-06 Method of screening for side effects of anticonceptives or estrogen and/or progesterone replacements or supplements

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EP0931263A1 true EP0931263A1 (de) 1999-07-28

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EP96930447A Withdrawn EP0931263A1 (de) 1996-09-06 1996-09-06 Screeningsferfahren für nebenwirkungen von antikonzeptionsmitteln oder östrogen und/oder progesteron ersatz- oder zusatzmitteln

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EP (1) EP0931263A1 (de)
JP (1) JP2002505736A (de)
AU (1) AU733553B2 (de)
CA (1) CA2265516C (de)
WO (1) WO1998010293A1 (de)

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Publication number Priority date Publication date Assignee Title
WO2001094951A2 (en) * 2000-06-08 2001-12-13 Board Of Regents, The University Of Texas System Inhibitors of c-reactive protein induced inflammation
KR101061562B1 (ko) 2008-08-28 2011-09-06 대한민국 비스테로이드성 항염제의 부작용을 예측하는 방법
ES2885523T3 (es) 2011-11-23 2021-12-14 Therapeuticsmd Inc Formulaciones y terapias de reposición hormonal de combinación naturales
US9301920B2 (en) 2012-06-18 2016-04-05 Therapeuticsmd, Inc. Natural combination hormone replacement formulations and therapies
US20130338122A1 (en) 2012-06-18 2013-12-19 Therapeuticsmd, Inc. Transdermal hormone replacement therapies
US20150196640A1 (en) 2012-06-18 2015-07-16 Therapeuticsmd, Inc. Progesterone formulations having a desirable pk profile
US10806740B2 (en) 2012-06-18 2020-10-20 Therapeuticsmd, Inc. Natural combination hormone replacement formulations and therapies
US10806697B2 (en) 2012-12-21 2020-10-20 Therapeuticsmd, Inc. Vaginal inserted estradiol pharmaceutical compositions and methods
US10568891B2 (en) 2012-12-21 2020-02-25 Therapeuticsmd, Inc. Vaginal inserted estradiol pharmaceutical compositions and methods
US9180091B2 (en) 2012-12-21 2015-11-10 Therapeuticsmd, Inc. Soluble estradiol capsule for vaginal insertion
US10471072B2 (en) 2012-12-21 2019-11-12 Therapeuticsmd, Inc. Vaginal inserted estradiol pharmaceutical compositions and methods
US11266661B2 (en) 2012-12-21 2022-03-08 Therapeuticsmd, Inc. Vaginal inserted estradiol pharmaceutical compositions and methods
US10537581B2 (en) 2012-12-21 2020-01-21 Therapeuticsmd, Inc. Vaginal inserted estradiol pharmaceutical compositions and methods
US11246875B2 (en) 2012-12-21 2022-02-15 Therapeuticsmd, Inc. Vaginal inserted estradiol pharmaceutical compositions and methods
CA2947767A1 (en) 2014-05-22 2015-11-26 Therapeuticsmd, Inc. Natural combination hormone replacement formulations and therapies
AU2015296609A1 (en) 2014-07-29 2016-12-22 Therapeuticsmd, Inc. Transdermal cream
CA3012985A1 (en) 2015-01-27 2016-08-04 Kardiatonos, Inc. Biomarkers of vascular disease
US10328087B2 (en) 2015-07-23 2019-06-25 Therapeuticsmd, Inc. Formulations for solubilizing hormones
AU2017239645A1 (en) 2016-04-01 2018-10-18 Therapeuticsmd, Inc. Steroid hormone pharmaceutical composition
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CA2265516C (en) 2004-02-03
AU733553B2 (en) 2001-05-17
AU6947596A (en) 1998-03-26
WO1998010293A1 (en) 1998-03-12
CA2265516A1 (en) 1998-03-12
JP2002505736A (ja) 2002-02-19

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