EP0651802A1 - Polypeptide mit serotoninerger (5ht6) rezeptor-aktivität, für sie kodierende nukleinsäuren und ihre verwendung - Google Patents

Polypeptide mit serotoninerger (5ht6) rezeptor-aktivität, für sie kodierende nukleinsäuren und ihre verwendung

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Publication number
EP0651802A1
EP0651802A1 EP93914777A EP93914777A EP0651802A1 EP 0651802 A1 EP0651802 A1 EP 0651802A1 EP 93914777 A EP93914777 A EP 93914777A EP 93914777 A EP93914777 A EP 93914777A EP 0651802 A1 EP0651802 A1 EP 0651802A1
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EP
European Patent Office
Prior art keywords
polypeptide
polypeptides
leu
sequence
ile
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Withdrawn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
EP93914777A
Other languages
English (en)
French (fr)
Inventor
Nourdine Amlaiky
Ursula Boschert
René HEN
Jean-Luc Plassat
Sylvie Ramboz
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.)
Institut National de la Sante et de la Recherche Medicale INSERM
Original Assignee
Institut National de la Sante et de la Recherche Medicale INSERM
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.)
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Publication date
Application filed by Institut National de la Sante et de la Recherche Medicale INSERM filed Critical Institut National de la Sante et de la Recherche Medicale INSERM
Publication of EP0651802A1 publication Critical patent/EP0651802A1/de
Withdrawn legal-status Critical Current

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides

Definitions

  • the present invention relates to new polypeptides and the genetic material allowing their expression. More particularly, it relates to new polypeptides having a serotonergic receptor activity.
  • Serotonin is a neuromodulator capable of inducing and modulating a wide variety of behaviors such as sleep, appetite, locomotion, sexual activity or even vascular contraction. It is recognized that the activity of 0 serotonin is mediated by its interaction with receptors, designated serotonergic receptors or 5-HT receptors (for 5-hydroxytryptamine). Molecular biology studies as well as pharmacological studies have revealed that there are a large number of 5-HT receptor subtypes. The 5-HT receptors which have been described to date belong either to the family of receptors linked to 5 ion channels (5-HT3 receptors), or to the family of receptors which interact with G proteins and which possess seven transmembrane domains.
  • 5-HT receptors which have been described to date belong either to the family of receptors linked to 5 ion channels (5-HT3 receptors), or to the family of receptors which interact with G proteins and which possess seven transmembrane domains.
  • the 5-HT receptors interacting with G proteins can be subdivided into two distinct groups: the 5HT1 receptors, comprising the mammalian subtypes 5HT1A, 0 5HT1B and 5HT1D as well as three 5HT Drosophila receptors; and 5HT2 receptors including the 5HT2 and 5HT1C subtypes.
  • 5HT4 receptors are probably not the only 5HT receptors in existence, since pharmacological studies have revealed other subtypes such as the 5HT4 receptors as well as certain receptors related to the 5HT1 5 subtype ("5HT1 like" receptors ). In addition, additional molecular biology studies have also revealed heterogeneities within the 5HT1B / 1D subtypes.
  • the present invention results from the discovery of new polypeptides having a serotonergic receptor activity. Although belonging to 0 the family of receptors which interact with G proteins, these new polypeptides differ from the serotonergic receptors already described (5HT1, 5HT2, 5HT3 and 5HT4) from the structural point of view as from the pharmacological point of view. More particularly, the invention results from the isolation and characterization of these new polypeptides, designated 5HT6, as well as genetic material allowing their expression or their identification.
  • a first object of the invention therefore resides in polypeptides comprising all or part of the peptide sequence SEQ ID No. 1 or a derivative thereof.
  • the term derivative designates any molecule obtained by modification of genetic and / or chemical nature of the peptide sequence SEQ ID n ° 1.
  • modification of genetic and / or chemical nature one can hear any mutation, substitution , deletion, addition and / or modification of one or more residues.
  • Such derivatives can be generated for different purposes, such as in particular that of increasing the affinity of the peptide for its ligand (s), that of improving its production levels, that of increasing its resistance to proteases, that of increasing and / or modifying its activity, or that of giving it new pharmacokinetic and / or biological properties.
  • derivatives resulting from an addition there may be mentioned, for example, chimeric polypeptides comprising an additional heterologous part linked at one end.
  • the term derivative also includes polypeptides homologous to polypeptide SEQ ID No. 1, derived from other cellular sources and in particular from cells of human origin, or from other organisms, and having an activity of the same type. Such homologous polypeptides can be obtained by hybridization experiments as described in the examples (see SEQ ID No. 4).
  • the polypeptides of the invention are polypeptides having the capacity to bind serotonin. Even more preferably, they are polypeptides having a serotonergic receptor activity. Still according to a preferred mode, the polypeptides of the invention are capable of being recognized by antibodies recognizing the complete peptide sequence SEQ ID No. 1.
  • a particular embodiment of the invention is represented by the 5HT6 polypeptide comprising the entire peptide sequence SEQ ID No. 1. As indicated in the examples, this polypeptide can be expressed in different cell types to form a functional serotonergic receptor.
  • polypeptides of the invention can be obtained by expression in a cellular host of a nucleotide sequence as described below, by synthesis chemical, based on sequences SEQ ID No. 1 or 4 using techniques known to those skilled in the art, or by a combination of these techniques.
  • polypeptides of the invention as defined above are designated by 5HT6 polypeptides.
  • the present invention also relates to any nucleotide sequence coding for a 5HT6 polypeptide. More preferably, it is a sequence chosen from:
  • the different nucleotide sequences of the invention can be of artificial origin or not. They can be genomic sequences, cDNA, RNA, hybrid sequences or synthetic or semi-synthetic sequences. These sequences can be obtained for example by screening DNA libraries (cDNA library, genomic DNA library) by means of probes prepared on the basis of the sequence SEQ ID No. 1. Such libraries can be prepared for starting from cells of different origins by conventional molecular biology techniques known to those skilled in the art.
  • the nucleotide sequences of the invention can also be prepared by chemical synthesis, in particular according to the phosphoramidite method, or also by mixed methods including chemical or enzymatic modification of sequences obtained by screening of libraries.
  • the nucleotide sequences of the invention can be used for the production of the 5HT6 polypeptides as defined above.
  • the part coding for said polypeptide is generally placed under the control of signals allowing its expression in a cellular host.
  • the choice of these signals can vary depending on the cell host used.
  • the nucleotide sequences of the invention can be part of a vector, which can be autonomous or integrative replication. More particularly, autonomously replicating vectors can be prepared using autonomously replicating sequences in the chosen host. As regards integrative vectors, these can be prepared for example by using sequences homologous to certain regions of the host genome, allowing, by homologous recombination, the integration of the vector.
  • the cellular hosts which can be used for the production of the 5HT6 polypeptides of the invention by the recombinant route are both eukaryotic and prokaryotic hosts.
  • suitable eukaryotic hosts there may be mentioned animal cells, yeasts, or fungi.
  • yeasts mention may be made of yeasts of the genus Saccharomyces, Kluyveromyces, Pichia, Schwanniomyces, or Hansenula.
  • yeasts mention may be made of yeasts of the genus Saccharomyces, Kluyveromyces, Pichia, Schwanniomyces, or Hansenula.
  • animal cells mention may be made of COS, CHO, C127, NIH-3T3 cells, etc.
  • the mushrooms there may be mentioned more particularly Aspergillus ssp. or Trichoderma ssp.
  • prokaryotic hosts it is preferred to use the following bacteria Kcoli, Bacillus, or Streptomyces.
  • nucleotide sequences of the present invention can also be used in the pharmaceutical field, either for the production of antisense sequences which can be used in the context of gene therapy, or else for the production of probes allowing detection, by hybridization experiments, the expression of serotonergic receptors in biological samples and the detection of genetic anomalies (polymorphism, mutations) or outliers.
  • Antisense oligonucleotides are small oliogonucleotides, complementary to the strand coding for a given gene, and therefore capable of specifically hybridizing with transcribed mRNA, inhibiting its translation into protein.
  • the subject of the invention is therefore the antisense oligonucleotides capable of at least partially inhibiting the production of 5HT6 polypeptides as defined above.
  • Such oligonucleotides can consist of all or part of the nucleotide sequences defined above. They are generally sequences or fragments of sequences complementary to sequences coding for peptides of the invention.
  • Such oligonucleotides can be obtained from the sequence SEQ ID No. 1 or 4, by fragmentation, etc., or by chemical synthesis.
  • the invention also allows the production of nucleotide probes, synthetic or not, capable of hybridizing with the nucleotide sequences defined above which code for 5HT6 polypeptides of the invention, or with the corresponding mRNAs.
  • probes can be used in vitro as a diagnostic tool, for the detection of the expression of a 5HT6 serotonergic receptor, or even for the detection of genetic anomalies (poor splicing, polymorphism, point mutations, etc.). Given the multiple activities of serotonin, the probes of the invention can thus make it possible to identify neurological, cardiovascular or psychiatric conditions as being linked to the 5HT6 receptors.
  • probes can also be used for the detection and isolation of homologous nucleic acid sequences coding for 5HT6 polypeptides as defined above, from other cellular sources and preferably from cells of human origin, as well as illustrated in the examples.
  • the probes of the invention generally contain at least 10 bases, and they can contain up to the entire sequence SEQ ID No. 1 or 4 or their complementary strand. Preferably, these probes are, prior to their use, marked. For this, different techniques known to those skilled in the art can be used (radioactive, enzymatic labeling, etc.). The hybridization conditions under which these probes can be used are indicated in the general cloning techniques below as well as in the examples.
  • Another subject of the invention relates to recombinant cells capable of expressing on their surface a 5HT6 polypeptide as defined above. These cells can be obtained by introducing a nucleotide sequence as defined above coding for a polypeptide of the invention, then culturing said cells under conditions of expression of said sequence.
  • the recombinant cells according to the invention can be both eukaryotic and prokaryotic cells.
  • eukaryotic cells which are suitable, mention may be made of animal cells, yeasts, or fungi.
  • yeasts mention may be made of yeasts of the genus Saccharomyce, Kluyveromyces, Pichia, Schwanniomyce, or Hansenula.
  • animal cells mention may be made of COS, CHO, C127, NIH-3T3 cells, etc.
  • the mushrooms there may be mentioned more particularly Aspergillus ssp. or Trichoderma ssp.
  • prokaryotic cells it is preferred to use the following bacteria E.coli, Bacillus, or Streptomyces.
  • the cells thus obtained can be used to measure the capacity of different molecules to behave as a ligand or as a modulator of the activity of the polypeptides of the invention. More particularly, they can thus be used in a process for highlighting and isolating ligands or modulators of the activity of the polypeptides of the invention, and, more preferably, of agonists and antagonists of serotonin.
  • Another object of the invention therefore relates to a process for detecting and / or isolating ligands of the 5HT6 polypeptides of the invention, according to which the following steps are carried out:
  • a molecule or a mixture containing different molecules, possibly unidentified is brought into contact with a recombinant cell as described above expressing on its surface a polypeptide of the invention under conditions allowing the interaction between said polypeptide of the invention and said molecule in the event that it has an affinity for said polypeptide, and,
  • this method of the invention is suitable for the detection and / or isolation of agonists and antagonists of serotonin for the 5HT6 polypeptides.
  • Another object of the invention relates to a method for detecting and / or isolating modulators of the 5HT6 polypeptides of the invention, according to which the following steps are carried out:
  • a molecule or mixture containing different molecules, possibly unidentified is brought into contact with a recombinant cell as described above, expressing on its surface a polypeptide of the invention, in the presence of 5HT, under conditions allowing interaction between said polypeptide of the invention and 5HT, and,
  • Another object of the invention relates to the use of a ligand or of a modulator identified and / or obtained according to the process described above as a medicament.
  • ligands or modulators can indeed make it possible to treat certain neurological, cardiovascular or psychiatric affections linked to the 5HT6 receptors.
  • the invention also relates to any medicament comprising as active principle at least one molecule acting on a 5HT6 polypeptide of the invention.
  • the molecule is a ligand or a modulator identified and / or isolated according to the method described above.
  • SEQ ID No. 1 Nucleotide and peptide sequences of the murine 5HT6 receptor.
  • the 1558 bp cDNA was sequenced on both strands from the EcoRI site to the Xhol site. The first 92 nucleotides are not shown.
  • Figure 2 Percentages of peptide sequence homology between the 5HT6 receptor presented SEQ ID No. 1 and other receptors of the family of receptors coupled to G proteins. The homologies were calculated on the conserved sequences: the transmembrane domain and its connection loops.
  • Figure 3 Saturation curve of [ 125 I] -LSD to the membranes of Cos-7 cells expressing the 5HT6 receptor. The membranes were incubated with ligand concentrations ranging from 50 pM to 1.25 nM, with or without 10 ⁇ M of 5HT. The specific link is shown. The insert represents the Scatchard analysis of the results.
  • Figure 4 Demonstration of homologous sequences by PCR on total RNA (1 ⁇ g) of different tissues.
  • Table 1 Pharmacological profile of the 5HT6 receptor. The results correspond to competitive experiments for the binding of [ 125 I] -LSD to the membranes of Cos-7 cells expressing the 5HT6 receptor transiently.
  • the numbers in parentheses correspond to the number of independent experiments carried out, each point being carried out in triplicate.
  • the filling of the prominent 5 ′ ends is carried out by the Klenow fragment of DNA Polymerase I of E. coli (Biolabs) according to the supplier's specifications.
  • the destruction of the protruding 3 ′ ends is carried out in the presence of the DNA polymerase of phage T4 (Biolabs) used according to the manufacturer's recommendations.
  • the destruction of the protruding 5 ′ ends is carried out by gentle treatment with nuclease SI. Mutagenesis directed in vitro by synthetic oligodeoxynucleotides is carried out according to the method developed by Taylor et al. [Nucleic Acids Res. 13_ (1985) 8749-8764] using the kit distributed by Amersham.
  • PCR Polymerase-catalyzed Chain Reaction, Saiki RK et al., Science 230 (1985) 1350-1354; Mullis KB and Faloona FA, Meth. ⁇ nzym. JL55_ (1987) 335-350] is carried out using a "DNA thermal cycler" (Perkin ⁇ lmer Cetus) according to the manufacturer's specifications. Verification of the nucleotide sequences is carried out by the method developed by Sanger et al. [Proc. Natl. Acad. Sci. USA, 74 (1977) 5463-5467] using the kit distributed by Amersham.
  • the normal stringency conditions are generally as follows: hybridization: 3 x SCC in the presence of 5 x Denhart's at 65 ° C; washing: 0.5 x SSC at 65 ° C.
  • This probe was used to screen a rat brain cDNA library constructed in the phage UniZap (Stratagene), under conditions of low stringency (formamide 30, 5 ⁇ SSC, 42 ° C). Among the positive phages obtained, one of them, weakly hybridizing to the probe, was isolated. This phage, called ⁇ SR and carried by the plasmid pSR, contained an insert of approximately 1.6 kb which was then introduced into the plasmid Bluescript. The sequence of this fragment was determined on the 2 strands using the dideoxynucleotide technique using synthetic oligonucleotides. The sequence thus obtained is presented in SEQ ID No. 1. It shows that the isolated cDNA carries an open reading phase of 367 amino acids.
  • hydrophobicity analysis shows that this protein carries seven hydrophobic domains, a peculiarity encountered in members of the family of receptors coupled to G proteins.
  • the N-terminal end also contains 2 N-glycosylation sites , and the suspected cytoplasmic domain contains the consensus sites of phosphorylation by protein kinases C and A.
  • the sequence of the 5HT6 receptor isolated above was compared with the sequences of the receptors coupled to the following G proteins: S31, 5HT1BB, 5HTlD ⁇ , 5HT1A, 5HT-dro2A, 5HT-drol. 5HT1C and 5HT2. These experiments revealed some homology in the potential transmembrane domain and in the connection loops, but not in the terminal regions or in the third cytoplasmic loop. Figure 2 gives the% homology at the level of the conserved regions.
  • the homolgia in the conserved regions, with the known receptors is weak, the best result being obtained with the serotonergic receptors 5HT1BB and 5HTlD ⁇ (54% homology), and with the receptor S31 which is not yet characterized.
  • Example 1 The cDNA fragment isolated in Example 1 was inserted into a eukaryotic expression vector, which was used to transfect Cos-7 cells. The membranes of the transfected cells obtained were then prepared and tested for their capacity to bind certain labeled serotonergic ligands.
  • the 1.6 kb cDNA coding for the 5HT6 receptor was isolated from the plasmid pSR in the form of an EcoRI-XhoI fragment, then inserted at the corresponding sites of the vector p513.
  • the vector p513 is derived from the vector pSG5 [Green et al., Nucl. Acids Res. l ⁇ . (1988) 369] by adding a multisite of cloning.
  • the recombinant vector thus obtained, designated p513SR was then used (20 ⁇ g per 10 cm plate) to transfect the Cos-7 cells in the presence of calcium phosphate.
  • the recombinant cells are harvested and the membranes are prepared according to the technique described by Amlaiky and Caron [J. Biol. Chem. 260 (1985) 1983]. Saturation binding and competition experiments were then carried out on these membranes in the presence of the following radiolabelled ligands: [ 125 I] -LSD; [ 12 5l] -cyanopindolol; [ 3 H] -8-OH-DPAT and [ 3 H] - spiperone. For this, the membrane samples (10-20 ⁇ g of protein) were incubated for 10 minutes at 37 ° C. in the presence of the ligand in a final volume of 250 ⁇ l of 50 mM Tris-HCl buffer (pH 7.4).
  • the reaction is then stopped by vacuum filtration on Whatman GF / C glass fiber filters, and rinsing 4 times with 4 ml of 50 mM Tris-HCl buffer (pH 7.4).
  • the non-specific binding was determined in the presence of 10 ⁇ M of 5HT. Radioactivity was measured with a ⁇ counter.
  • [ 3 H] -8-OH-DPAT and [ 3 H] -s ⁇ iperone do not bind the prepared membranes
  • the [ 125 I] -LSD did not bind the Cos-7 cells transfected with the plasmid p513.
  • the cDNA cloned in Example 1 was also expressed in NIH-3T3 cells, which do not express any serotonergic receptor endogenously.
  • the recombinant expression vector described in 3. above was used. It was introduced (20 ⁇ g per 10 cm plate) into NIH-3T3 cells by transfection in the presence of calcium phosphate, at the same time as the vector pRSVneo [Gorman et al., Science 221 (1983) 551], carrying the G418 resistance gene (1 ⁇ g per 10 cm plate).
  • the transforming clones were selected in the presence of 0.5 mg of G418.
  • the isolated clones were then amplified and the total RNAs of these clones were prepared and analyzed in Northern Blot for the expression of 5HT6 mRNA.
  • a clone was thus selected, SR4, expressing high levels of 5HT6 mRNA.
  • the cell membranes of this clone were then prepared and tested under the conditions described above for their capacity to bind certain ligands. labeled serotonergic, testifying to the presence of functional 5HT6 receptors on their surface.
  • the nucleotide sequence SEQ ID No. 1 was then used to demonstrate homologous sequences from other tissues. For this, two techniques were used:
  • tissues used for the search for homologous sequences are the following of murine origin: brain, cerebellum, kidney, liver, spinal cord, spleen, lung, intestine and heart.
  • the probe (i) corresponds to position 1174 on SEQ LD n ° 1 and the probe ( ⁇ ) at position 1394.
  • RNAs were prepared from the various tissues studied, using the technique described by Cathala et al. (DNA 2 (4) (1983)). 1 ⁇ g of these RNAs was subjected to reverse transcription in the presence of 200 units of MMLV reverse transcriptase and 300 ng of probe (i), for 1 hour at 37 ° C. Half of the product of this reaction was then amplified (20 cycles) in the presence of 5 units of the Taq polymerase (Cetus) and 500 ng of the probes (i) and (ii).
  • the in situ hybridization experiments were carried out on cryostatic sections of the adult rat brain (approximately 8 weeks) according to the technique described by Col et al. [EMBO J. 2 (1983) 617].
  • the probe used for these experiments is a single-stranded RNA obtained by transcription in the presence of T7 polymerase,
  • a human genomic DNA library was prepared from the placenta, by partial digestion with the enzyme Mbol, separation on salt gradients, and under cloning in the vector Lamda GEM 12 linearized by BamHI (host bacterium: TAP 90).
  • the library thus obtained was then screened using the 1.6 kb EcoRI-XhoI fragment described in example 3 as a probe, labeled according to the random priming technique.
  • the DNA fragments which hybridize with this probe were isolated, subcloned in a Bluescript plasmid, amplified, then sequenced in both directions according to the dideoxynucleotide technique.
  • the amplification was carried out by the PCR technique: 20 cycles in the presence of Thermus aquaticus polymerase (2.5 units; Cetus) and of oligonucleotides 1 (SEQ ID No. 5) and 2 (SEQ ID No. 6).
  • the fragment obtained was digested with the enzymes BamHI and Xhol, then subcloned into an expression vector P513.
  • sequence obtained is presented on the sequence SEQ ID No. 4. It is understood that the same experiments can be repeated using other tissues and in particular tissues of human origin, and other probes. Furthermore, the homologous sequences demonstrated during these experiments can obviously be then isolated and / or amplified by conventional techniques of molecular biology.
  • AAG ACA TTA TAC CAC AAG AGA CAA GCA AGT AGG ATT GCA AAG GAG GAG 672 Lys Thr Leu Tyr His Lys Arg Gin Ala Ser Arg Ile Ala Lys Glu Glu 210 215 220

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EP93914777A 1992-07-01 1993-06-29 Polypeptide mit serotoninerger (5ht6) rezeptor-aktivität, für sie kodierende nukleinsäuren und ihre verwendung Withdrawn EP0651802A1 (de)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
FR9208082 1992-07-01
FR9208082A FR2693201B1 (fr) 1992-07-01 1992-07-01 Nouveaux polypeptides ayant une activité de récepteur sérotoninergique, acides nucléiques codant pour ces polyptides et utilisations.
PCT/FR1993/000651 WO1994001556A1 (fr) 1992-07-01 1993-06-29 Polypeptides ayant une activite de recepteur serotoninergique (5ht6), acides nucleiques codant pour ces polypeptides et utilisations

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EP0651802A1 true EP0651802A1 (de) 1995-05-10

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EP93914777A Withdrawn EP0651802A1 (de) 1992-07-01 1993-06-29 Polypeptide mit serotoninerger (5ht6) rezeptor-aktivität, für sie kodierende nukleinsäuren und ihre verwendung

Country Status (5)

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EP (1) EP0651802A1 (de)
JP (1) JPH07508655A (de)
CA (1) CA2139431A1 (de)
FR (1) FR2693201B1 (de)
WO (1) WO1994001556A1 (de)

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Publication number Priority date Publication date Assignee Title
US6300087B1 (en) 1992-11-03 2001-10-09 Synaptic Pharmaceutical Corporation DNA encoding a human serotonin receptor (5-HT4B) and uses thereof
NZ258320A (en) 1992-11-03 1996-12-20 Synaptic Pharma Corp Dna encoding a human receptor 5-ht4b receptor and diagnostic use
CA2274055A1 (en) * 1996-12-19 1998-06-25 Smithkline Beecham P.L.C. N-piperazin-1-ylphenyl-benzamide derivatives

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5155218A (en) * 1990-05-08 1992-10-13 Neurogenetic Corporation Dna encoding human 5-ht1d receptors
DE4041464A1 (de) * 1990-12-22 1992-06-25 Basf Ag 5-ht(pfeil abwaerts)1(pfeil abwaerts)-rezeptor

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See references of WO9401556A1 *

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CA2139431A1 (fr) 1994-01-20
WO1994001556A1 (fr) 1994-01-20
FR2693201B1 (fr) 1994-08-19
JPH07508655A (ja) 1995-09-28
FR2693201A1 (fr) 1994-01-07

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