US20210317156A1

US20210317156A1 - New polymer linked multimers of guanosine-3', 5'-cyclic monophosphates

Info

Publication number: US20210317156A1
Application number: US16/329,031
Authority: US
Inventors: Hans-Gottfried Genieser; Frank Schwede; Andreas Rentsch; Valeria Marigo
Original assignee: Mireca Medicines GmbH
Current assignee: Graybug Vision Inc
Priority date: 2016-08-31
Filing date: 2017-08-31
Publication date: 2021-10-14
Also published as: JP2019526576A; JP2023102288A; JP7298048B2; EP3507295A1; WO2018041942A1; US20230242568A1; WO2018041942A9

Abstract

Embodiments of the invention are directed to new polymer linked multimeric guanosine-3′, 5′-cyclic monophosphate (cGMP) analogues that modulate the cGMP-signaling system, preferably having activating properties, and more preferably being activators of cGMP dependent protein kinase (PKG), and related monomeric precursors thereof. The invention is also directed to related monomeric compounds, which may also show modulating activity and/or may serve as monomeric precursors of the multimers. The invention further relates to the use of such compounds as reagents for signal transduction research and as modulators of cyclic nucleotide-regulated binding proteins and isoenzymes thereof, and as ligands for affinity chromatography, for antibody production or for diagnostic applications e.g. on chip surfaces.

Description

FIELD OF THE INVENTION

The present invention relates to novel polymer linked multimers of guanosine-3′, 5′-cyclic nucleotide monophosphates, including tethered di-, tri- and tetramers and their application in the fields of medicine and pharmacy. The invention also relates to specific monomers as precursors. The invention further relates to the use of such compounds as reagents for signal transduction research and as modulators of cyclic nucleotide-regulated binding proteins and isoenzymes thereof, and as ligands for affinity chromatography, for antibody production or for diagnostic applications e.g. on chip surfaces.

BACKGROUND OF THE INVENTION

Guanosine-3′,5′-cyclic monophosphate (cGMP) is a purine nucleobase-containing cyclic nucleotide and was discovered as endogenous molecule in 1963. It is well known to act as a second messenger, wherein its intracellular level is altered as a response to (primary) signaling molecules such as toxins, hormones or nitric oxide, which in turn induces diverse cellular processes, such as gene control, chemotaxis, proliferation, differentiation, and apoptosis. Several diseases like retinal degeneration, cardiovascular diseases, asthma or diabetes are associated with unusually high or low levels of cGMP.¹The cGMP signaling cascade therefore has been recognized as a potential pharmacological target and is investigated by numerous academic groups and pharmaceutical companies. Research in this field demands for compounds that effectively modulate different targets of the said cascade. For this purpose, a number of cGMP analogues featuring cell permeability (in contrast to cGMP), enhanced activity and increased resistance to degradation by phosphodiesterases (PDE) have been established.
However, for the use as a drug and/or research tool cGMP analogues need to comply with a complex profile of characteristics that is unique for each biological system, to achieve a maximum effect. While applied compounds are usually selected for their ability to interfere with the main target of a studied mechanism of a disease, condition or disorder, there are always several required characteristics of a compound, that cannot be predicted and demand for testing a large set of analogues. Accordingly, for the increasing number of applications there is growing need for constantly expanding the group of available cGMP analogues with derivatives that feature another combination of characteristics as well as further improved activation potential, target specificity or multi target effects. Also tailor-made modifications such as reporting groups are desired variations for instance for research or diagnostic purposes. One of the targets addressed by cGMP and its analogues is the cGMP dependent protein kinase (PKG) from which three isoforms (Iα, Iß and II) are known. Knowledge on the identity and presence of PKG substrates in different cells, tissues and organisms is restricted. Hence the physiological as well as pathological importance of the cGMP-PKG system is not well understood, which is likely to have reduced the general understanding of cGMP-related phenomena, as well as the development of therapies in diseases and conditions where such substrates are involved. Efficient and reliable PKG activators will make it possible to address this question much more sharply than what can be currently achieved.
Among the PKG isoforms compounds activating PKG Iα have been explored most widely, while far less derivatives targeting Iß and even fewer interfering with PKG II are available. In fact, only a small number of compounds were reported to feature PKG II activation.^{2 3}For biochemical assays the most potent and predominantly applied one is 8-pCPT-cGMP. A drawback of this compound, however, is associated with the substituent in the 8-position. The 8-pCPT-moiety has been reported to induce additional off target effects besides PKG activation.⁴Furthermore it is not suitable for assays involving UV light, as it is decomposed under these conditions. For some applications it is therefore desirable to have strong PKG II agonists that don't feature an 8-pCPT-group.
A desired selective biological effect is always also an issue of the activation potential. The less a compound activates a studied target, the larger amount of substance needs to be applied and thus the higher is the probability of off target effects such as extra cellular bindings.⁴In conclusion it is favorable to use agonists with improved activation potential, reasoning a constant need for such compounds.
A class of PKG agonists, which could offer superior activity, are polymer linked multimers (PLM) of cGMP analogues, including di-, tri- and tetramers. This concept comprises the idea of achieving a strong enhancement of activity through addressing multiple binding sites with a single molecule. Prior to the present invention, however, the concept had only been applied once on a dimeric compounds (tri- and tetrameric analogues have not been reported). Therein, just a single type of a polymer linked dimeric cGMP (PLD), or more precisely one homologous series, differing only in the spacer length, has been disclosed. The preparation of said homologues was described as a coupling via the 8-position by reacting 8-thio cGMP with bifunctional PEG vinyl sulfones (VS-PEG_n-VS) affording structures of general formula cGMP-8-S—(CH₂)₂—SO₂-PEG_n-SO₂—(CH₂)₂—S-8-cGMP (also see FIG. 1). Dependent on the length of the (PEG) spacer these PLDs displayed an increased activation potential for either PKG Iα (up to 30-fold) or cyclic-nucleotide-gated (CNG) channels (up to 1000-fold) compared to cGMP itself. The most potent commercially available monomeric PKG Iα agonist 8-(2-NH₂-Ph-S)-cGMP (8-APT-cGMP) in contrary exhibits only an about 15-fold higher activity than cGMP.⁶
Applicability of the PLM concept has been proposed for further targets,⁸but no concrete methods to generate PLMs with superior activating potential for PKG isoforms Iß and II were disclosed and no further coupling strategies or synthetic protocols were reported. Particularly the synthetic access, however, comprises a drawback of the reported PLDs. Applying the published protocol, the desired compound is obtained in poor yields only as a byproduct. This outcome results from a favored addition of vinyl sulfones to the 7-position of cGMP.⁷PLDs featuring a different coupling moiety, that supports a more reliable access as well as maintained or preferably increased PKG activation, are therefore needed.
In addition, compounds featuring a similarly increased PKG Iß and/or PKG II activation potential as described for Iα targeting PLDs above, would be valuable tools for numerous research groups. The effects on these isoforms, however, have never been studied in connection with polymer linked di-, tri- or tetramers. Accordingly it was desirable to provide polymer linked multimeric activators of PKG Iß and II, but unknown, whether the concept could be transferred to these isoforms and if, what modifications would be necessary.
The effect of structural manipulations at the nucleobase in context with polymer linked multimers also has not been studied before. This in turn is especially important, though, as established compounds used as biochemical tools often require customized derivatization for specific applications. Among others introduction of a reporter moiety that facilitates a certain assay read out or detection method can be desirable. Coupling of the respective compound to a fluorescent dye, for instance, is a common strategy in this context. This dye in turn allows localizing the intracellular distribution of the compound and its binding proteins in living cells by means of microscopic or spectroscopic techniques. A frequently observed drawback of strategies that involve structural manipulation of an activator, however, is the change of nature of the parent compound. Even minor modifications can result in a significant shift of target affinity and specificity or even loss of activation potential.
In addition, compounds, which simultaneously activate multiple targets such as two or all three PKG isoforms, could be beneficial for some applications and unsolved problems in signal transduction research.
Supported by the results of the only homologoues PLD series reported before (as stated above), however, the enhanced target activation of these compounds appeared to fundamentally depend on an optimum spacer length (between the cGMP units), which is unique for each addressed protein. Therefore, addressing two or more targets, with the same PLD, seemed, if feasible at all, only possible with an intermediate spacer length at which the activation potential for both targets would be significantly decreased.
Accordingly, there is a growing need for activators of the cGMP signaling cascade, tailor-made for individual biochemical applications, featuring superior single or multiple target activation, with or without additional functionalization that for instance facilitates specific assay read outs. To combine all these features within a single monomeric structure can be very difficult to accomplish. On such relatively small compounds each of the multiple modifications necessary can have a significant influence on the original target binding properties.
The situation, however, changes when it comes to multimers, wherein each desired modification can be introduced on a different cGMP unit. Thus, the respective impact on the biochemical activation profile follows different rules. PLMs offer access to a highly potent, so far rarely studied class of compounds. The challenges of transferring their potential to further targets of the cGMP signal transduction cascade (in particular PKG Iß and II) and addressing multiple targets simultaneously, while optionally featuring a reporting group, though, have not been mastered yet.

OBJECTIVE AND SUMMARY OF THE INVENTION

Therefore the achieved objective of the present invention has been to provide new polymer linked multimeric analogues of guanosine-3′,5′-cyclic monophosphate (PLMs) including di-, tri- and tetrameric derivatives, which are compared to monomeric analogues used so far, improved in terms of PKG Iß and/or PKG II activation potential. Preferably the new PLMs additionally interfere with PKG Iα, wherein it is even more preferred that the activation potential for this isoform exceeds the previously reported one for a polymer linked dimer.⁵Another objective of the invention has been, to provide PKG activators, which can be functionalized (e.g. with a reporting group), while essentially maintaining their target affinity. A further objective of the invention has been to provide the new PLMs as pharmaceutically acceptable analogues for treating or diagnosing a disease, condition or disorder associated with dysregulation of a cGMP-effected cellular target, wherein additional targets can be, including, but not limited to, a hyperpolarization-activated cyclic nucleotide-gated (HCN) channel, a phosphodiesterase (PDE) and a cGMP-gated channel (CNGC). In another aspect, the objective of the invention has been to provide the new PLMs for application as research tool to identify and validate the cGMP-system in cell cultures or tissues or as a diagnostic tool. Embodiments of the invention are directed to new polymer linked multimeric guanosine-3′, 5′-cyclic monophosphate (cGMP) analogues that modulate the cGMP-signaling system, preferably having activating properties, and more preferably being activators of cGMP dependent protein kinase (PKG), and related monomeric precursors thereof.
Another achieved objective of the present invention has been to provide related monomeric compounds, which may serve as monomeric precursors of the multimers of the present invention and/or may also show modulating activity. Thus, another objective of the invention has been to provide related monomeric compounds as pharmaceutically acceptable analogues for treating or diagnosing a disease, condition or disorder associated with dysregulation of a cGMP-effected cellular target, wherein additional tar-gets can be, including, but not limited to, a hyperpolarization-activated cyclic nucleotide-gated (HCN) channel, a phosphodiesterase (PDE) and a cGMP-gated channel (CNGC). In another aspect, the objective of the invention has been to provide the new related monomeric compounds for application as research tool to identify and validate the cGMP-system in cell cultures or tissues or as a diagnostic tool. Embodiments of the invention are directed to new related monomeric compound analogues that modulate the cGMP-signaling system, preferably having activating properties, and more preferably being activators of cGMP dependent protein kinase (PKG).

BRIEF DESCRIPTION OF THE FIGURES AND FORMULAS

Formula I General constitution of compounds of the invention (branched and linear analogues).

Formula Ib More detailed illustration of Formula I.

Formula II General constitution of compounds of the invention (linear analogues).

Formula IIb More detailed illustration of Formula II.

Formula III General constitution of G units as discrete compounds of the invention or units of compounds according to Formula I or II.

Formula IV and V General constitution of G units according to Formula III, featuring exemplary imidazolinone substitution.

FIG. 1 Previously reported polymer linked dimeric cGMP⁵

Legend: Guanosine-3′, 5′-cyclic monophosphate-[8-thioethylsulfonyl-(ethyloxy)_n-ethylsulfonylethylthio-8]-guanosine-3′, 5′-cyclic monophosphate (polydispers compound arising from synthesis with polydispers bis vinylsulfonyl-PEG_nwith M_wof 800 g/mol, 1.2 kg/mol, 3.4 kg/mol or 20 kg/mol; or monodispers compound with n=6).

FIG. 2 Example of a trimeric compound according to the invention, illustrating the used variables.

FIGS. 3 to 5 In vitro activation of PKG isoforms by polymer linked cGMP derivatives featuring different spacer lengths with and without PET-modification (FIG. 3), varied linking position (FIG. 4) and unequal cGMP (analogue) units with and without unequal linking positions (FIG. 5).

Legend: PKG isozymes Iα (0.2 nM), Iβ (0.15 nM) and II (0.5 nM) were incubated with different concentrations (10 pM to 6 μM) of compounds of the invention and cGMP as reference compound at room temperature for 60 min. The activation values of the compounds are expressed as relative PKG activation compared to cGMP with cGMP set as 1 for each kinase isozyme. The K_a-values of cGMP for half-maximal kinase activation were 28 nM for Iα, 425 nM for Iβ and 208 nM for II. Compound numbers refer to analogues displayed in Table 13.

FIG. 6 Expression of PKG isoforms in 661W cells.

Legend: RT-PCR on cDNA from mRNA extracted from 661W cell. The 661W cell line expresses the PKG isoforms Iα and II. Heart and muscle tissues were used as positive controls.

FIG. 7 Increased cell death in the 661W cell line after treatment with different polymer linked dimeric cGMP analogues.

Legend: 661W cells were exposed to compounds for 16 hours at different concentrations (1 nM to 10 μM) and percentage of dying cells was evaluated by Ethidium Homodimer assay. Untreated cells are shown as control sample (black bar). Reference compound 8-Br-PET-cGMP is shown as dashed bars. Data are presented as means±SD from at least three biological replicates. Results not including standard deviation refer to single measurements. Asterisks indicate the P value of the unpaired Student's t-test (*P≤0.05, **P≤0.01, ***P≤0.001), statistically assessing significant differences between untreated and treated cells, wherein a p value ≤0.05 was considered significant. Compound numbers refer to analogues displayed in Table 13.

SPECIFICATION OF STRUCTURES

The new polymer linked multimeric cGMP analogues of the invention are compounds having the formula (I) or (II)
wherein:
G units G¹and G²independently are compounds of formula (III) and G units G³and G⁴independently from G¹and G²and independently from each other are compounds of formula (III) or absent, wherein in case of formula (II) G⁴is always absent if G³is absent,
and wherein in formula (III)
X, Y and Z are N
R₁, R₄, R₅, R₇and R₈independently can be equal or individual for each G unit (G¹, G², G³and G⁴), while

- R₁can independently be H, halogen, azido, cyano, acyl, aracyl, nitro, alkyl, aryl, aralkyl, OH, O-alkyl, O-aryl, O-aralkyl, O-acyl, O-aracyl, SH, S-alkyl, S-aryl, S-aralkyl, S-acyl, S-aracyl, S(O)alkyl, S(O)-aryl, S(O)-aralkyl, S(O)-acyl, S(O)-aracyl, S(O)₂-alkyl, S(O)₂-aryl, S(O)₂-aralkyl, S(O)₂-acyl, S(O)₂-aracyl, SeH, Se-alkyl, Se-aryl, Se-aralkyl, NR9R10, SiR13R14R15 wherein R9, R10, R13, R14, R15 independently from each other can be H, alkyl, aryl, aralkyl;
- R₂is absent;
- R₃is OH;
- R₄can independently be absent, H, amino, alkyl, aralkyl, nitro, N-oxide, or can form together with R₃, Y and the carbon bridging Y and R₃an imidazole ring which can be unsubstituted or substituted with alkyl, aryl or aralkyl, or can form together with Y and R₅and the carbon bridging Y and R₅an imidazole ring which can be unsubstituted or substituted with alkyl, aryl or aralkyl, or can form together with Y and R₅and the carbon bridging Y and R₅an imidazolinone as depicted (structure IV, V, n=1) or an homologous ring (n=2 to 8) which each can be unsubstituted or substituted (not depicted) with alkyl, aryl or aralkyl;

- R₅can independently be H, halogen, NR30R31, NH-carbamoylR32R33 wherein R30, R31, R32, R33, independently from each other can be H, alkyl, aryl, aralkyl, or can form together with R₄, Y and the carbon bridging Y and R₅an imidazole ring which can be unsubstituted or substituted with alkyl, aryl or aralkyl, or can form together with Ra, Y and the carbon bridging Y and R₅an imidazolinone ring as depicted (structure IV, V, n=1) or an homologous ring (n=2 to 8) which each can be unsubstituted or substituted (not depicted) with alkyl, aryl or aralkyl;
- R₆is OH;
- R₇is O-carbamoyl-alkyl, O-carbamoyl-aryl, O-carbamoyl-aralkyl, OH, O-alkyl, O-aryl, O-aralkyl, O-acyl, SH, S-alkyl, S-aryl, S-aralkyl, borano (BH₃), methylborano, dimethylborano, cyanoborano (BH₂CN), S-PAP, O-PAP, S-BAP, or O-BAP,
  - wherein PAP is a photo-activatable protecting group with non limiting examples of, optionally, PAP=o-nitro-benzyl, 1-(o-nitrophenyl)-ethylidene, 4,5-dimethoxy-2-nitrobenzyl, 7-dimethylamino-coumarin-4-yl (DMACM-caged), 7-diethylamino-coumarin-4-yl (DEACM-caged) and 6,7-bis(carboxymethoxy)coumarin-4-yl)methyl (BCMCM-caged);
  - and wherein BAP is a bio-activatable protecting group with non limiting examples of, optionally, BAP=methyl, acetoxymethyl, pivaloyloxymethyl, methoxymethyl, propionyloxymethyl, butyryloxymethyl, cyanoethyl, phenyl, benzyl, 4-acetoxybenzyl, 4-pivaloyloxybenzyl, 4-isobutyryloxybenzyl, 4-octanoyloxybenzyl, 4-benzoyloxybenzyl; and
- R₈is O-carbamoyl-alkyl, O-carbamoyl-aryl, O-carbamoyl-aralkyl, OH, O-alkyl, O-aryl, O-aralkyl, O-acyl, O-PAP or O-BAP,
  - wherein PAP is a photo-activatable protecting group with non limiting examples of, optionally, PAP=o-nitro-benzyl, 1-(o-nitrophenyl)-ethylidene, 4,5-dimethoxy-2-nitrobenzyl, 7-dimethylamino-coumarin-4-yl (DMACM-caged), 7-diethylamino-coumarin-4-yl (DEACM-caged) and 6,7-bis(carboxymethoxy)coumarin-4-yl)methyl (BCMCM-caged);
  - and wherein BAP is a bio-activatable protecting group with non limiting examples of, optionally, BAP=methyl, acetoxymethyl, pivaloyloxymethyl, methoxymethyl, propionyloxymethyl, butyryloxymethyl, cyanoethyl, phenyl, benzyl, 4-acetoxybenzyl, 4-pivaloyloxybenzyl, 4-isobutyryloxybenzyl, 4-octanoyloxybenzyl, 4-benzoyloxybenzyl;

and wherein
linking residues LR¹, LR², LR³and LR⁴independently can replace or covalently bind to any of the particular residues R₁, R₄and/or R₅of the G units (G^1-4) they connect,
wherein in case they bind to any of the residues R₁, R₄and/or R₅, an endstanding group of the particular residue (R₁, R₄and/or R₅), as defined above, is transformed or replaced in the process of establishing the connection and is then further defined as part of the particular linking residue (LR^1-4) within the assembled compound,

- while
- LR¹is (a) a tri- or tetravalent branched hydrocarbon moiety or (b) a divalent hydrocarbon moiety each with or without incorporated heteroatoms such as, but not limited to, O, N, S, Si, Se, B, wherein the backbone preferably contains 1 to 28 carbon atoms and can be saturated or unsaturated, substituted or unsubstituted,
  - while
  - each attachment point independently can be a substituted or unsubstituted carbon- or heteroatom
  - and
  - in case poly ethylene glycole (PEG) moieties are incorporated in accordance to the definition, the preferred number of carbon atoms can be exceeded by the number present in the PEG moieties, wherein all PEG moieties together can contain a total amount of
    - 1 to 500 ethylene glycol groups (—(CH₂CH₂O)_n— with n=1 to 500) in case of divalent linking residue (LR¹)
    - or
    - 1 to 750 ethylene glycol groups (—(CH₂CH₂O)_n— with n=1 to 750) in case of trivalent linking residue (LR¹)
    - or
    - 1 to 1000 ethylene glycol groups (—(CH₂CH₂O)_n— with n=1 to 1000) in case of tetravalent linking residue (LR¹),
  - and, if substituted,
  - substituents include, but are not limited to, optionally one or more alkyl groups, halogen atoms, haloalkyl groups, (un)substituted aryl groups, (un)substituted heteroaryl groups, amino, oxo, nitro, cyano, azido, hydroxy, mercapto, keto, carboxy, carbamoyl, expoxy, methoxy, ethynyl,
  - and/or substituents can further be connected to each other, forming a ringsystem with 1 to 4 rings, with or without incorporated heteroatoms, saturated or unsaturated, substituted or unsubstituted, aliphatic or aromatic;
- LR², LR³and LR⁴are divalent hydrocarbon moieties with or without incorporated heteroatoms such as, but not limited to, optionally heteroatoms O, N, S, Si, Se, B, wherein the backbone preferably contains 1 to 28 carbon atoms and can be, saturated or unsaturated, substituted or unsubstituted,
  - while
  - each attachment point independently can be a substituted or unsubstituted carbon- or heteroatom
  - and
  - in case poly ethylene glycole (PEG) moieties are incorporated in accordance to the definition, the preferred number of carbon atoms can be exceeded by the number present in the PEG moieties, wherein all PEG moieties together can contain a total amount of 1 to 500 ethylene glycol groups (—(CH₂CH₂O)_n— with n=1 to 500)
  - and, if substituted,
  - substituents include, but are not limited to, optionally one or more alkyl groups, halogen atoms, haloalkyl groups, (un)substituted aryl groups, (un)substituted heteroaryl groups, amino, oxo, nitro, cyano, azido, hydroxy, mercapto, keto, carboxy, carbamoyl, expoxy, methoxy, ethynyl,
  - and/or substituents can further be connected to each other, forming a ringsystem with 1 to 4 rings, with or without incorporated heteroatoms, saturated or unsaturated, substituted or unsubstituted, aliphatic or aromatic;

wherein in case of formula (II) if G⁴is absent, LR⁴is absent, too, and
wherein in case of formula (II) if G³and G⁴are absent, LR³and LR⁴are absent, too,
and wherein
G¹, G², G³and G⁴can further be salts and/or hydrates

- while, optionally, non limiting examples of suitable salts of the particular phosphate moiety are lithium, sodium, potassium, calcium, magnesium, zinc or ammonium, and trialkylammonium, dialkylammonium, alkylammonium, e.g., triethylammonium, trimethylammonium, diethylammonium and octylammonium;

and wherein
G¹, G², G³and G⁴can optionally be isotopically or radioactively labeled, be PEGylated, immobilized or be labeled with a dye or another reporting group,

- wherein
- the reporting group(s) and/or dye(s)
  - (a) are coupled to G¹, G², G³and/or G⁴via a linking residue (LR⁵), bound covalently to or replacing any of the particular residues R₁, R₄and/or R₅independently for each G unit (G¹, G², G³and/or G⁴)
    - while LR⁵can be as defined for LR²
  - or
  - (b) in case of formula (I) can replace G³and/or G⁴
- and wherein
- examples of optionally suitable dyes include, but are not limited to, fluorescent dyes such as fluorescein, anthraniloyl, N-methylanthraniloyl, dansyl or the nitro-benzofurazanyl (NBD) system, rhodamine-based dyes such as Texas Red or TAMRA, cyanine dyes such as Cy™3, Cy™5, Cy™7, EVOblue™10, EVOblue™30, EVOblue™90, EVOblue™100 (EVOblue™-family), the BODIPY™-family, Alexa Fluor™-family, the DY-family, such as DY-547P1, DY-647P1, coumarines, acridines, oxazones, phenalenones, fluorescent proteins such as GFP, BFP and YFP, and near and far infrared dyes
- and wherein
- reporting groups optionally include, but are not limited to, quantum dots, biotin and tyrosylmethyl ester;
- and wherein
- PEGylated refers to the attachment of a single or multiple LR^PEGgroup(s) independently, wherein LR^PEGcan be as defined for LR², with the provisos that in this case (i) of LR²only one terminus is connected to a G unit (G¹, G², G³and/or G⁴) by covalently binding to or replacing any of the particular residues R₁, R₄, and/or R₅independently for each G unit (G¹, G², G³and/or G⁴), and (ii) the other terminus of LR²is either an alkyl group or a reactive group that allows for conjugation reactions and/or hydrogen bonding
  - while, optionally, non limiting examples of reactive groups are, —NH₂, —SH, —OH, —COOH, —N₃, —NHS-ester, halogen group, epoxide, ethynyl, allyl
- and with the proviso (iii) that LR^PEGhas incorporated ethylene glycol moieties (—(CH₂CH₂O)_n— with n=2 to 500)

with the proviso that the compound of formula (I) and/or (II), is not selected from structures depicted in FIG. 1; Guanosine-3′, 5′-cyclic monophosphate-[8-thioethylsulfonyl-(ethyloxy)n-ethylsulfonylethylthio-8]-guanosine-3′, 5′-cyclic monophosphate (polydispers compound arising from synthesis with polydispers bis vinylsulfonyl-PEGn with Mw of 800 g/mol, 1.2 kg/mol, 3.4 kg/mol or 20 kg/mol; or monodispers compound with n=6).
In a particular embodiment, the precedingly defined compound of formula (I) and/or (II) may be a compound, wherein at least two G units are unequally substituted.
In another particular embodiment, the precedingly defined compound of formula (I) and/or (II) may be a compound, wherein in case of formula (I) G3 and G4 are absent, or in case of formula (II) G³, G⁴, LR³and LR⁴are absent; and wherein R₄is not H and/or R₅is not NH₂.
In a particular variant embodiment, the precedingly defined compound of formula (I) and/or (II) may be a compound, wherein at least one G unit is linked via a position other than R₁.
In a further particular embodiment, the precedingly defined compound of formula (I) and/or (II) may be a compound, wherein in case of formula (I) G³and G⁴are absent, or in case of formula (II) G³, G⁴, LR³and LR⁴are absent.
In still another particular embodiment, any of the precedingly defined compounds of formula (I) and/or (II) may be a compound, wherein all R₇are SH and all R₈are O, or all R₇are O and all R₈are OH.
Any reasonable combination of the before embodiments is possible, too, according to the invention.

Chemical Definitions

Listed below are the definitions of various terms and phrases used to describe the compounds of the present invention. These definitions apply to the terms as they are used throughout the specification.
Halogen refers to F, Cl, Br, and I.
Alkyl refers to an alkyl group, which is a hydrocarbon moiety with 1 to 28, preferably 1 to 20 carbon atoms, with or without (integrated) heteroatoms such as but not limited to O, S, Si, N, Se, B, wherein the point of attachment unless specified otherwise is a carbon atom. Its constitution can be
Linear saturated hydrocarbon moiety—including, but not limited to, methyl, ethyl, propyl, butyl and pentyl or
Linear unsaturated hydrocarbon moiety—containing more preferably 2 to 20 carbon atoms, including, but not limited to, ethylen, propylen, butylen and pentylen or
Branched saturated hydrocarbon moiety—deviating from the general alkyl definition by containing at least 3 carbon atoms and including, but not limited to, isopropyl, sec.-butyl and tert.-butyl or
Branched unsaturated hydrocarbon moiety—deviating from the general alkyl definition by containing at least 3 carbon atoms and including, but not limited to, isopropenyl, isobutenyl, isopentenyl and 4-methyl-3-pentenyl
or
Cyclic saturated hydrocarbon moiety—containing more preferably 3 to 8 ring atoms and including, but not limited to, cyclopentyl, cyclohexyl, cycloheptyl, piperidino, piperazino
or
Cyclic unsaturated hydrocarbon moiety—containing more preferably 3 to 8 ring atoms.
Herein the term saturated means the group has no carbon-carbon double and no carbon-carbon triple bonds. However, in the substituted case of saturated groups one or more carbon-oxygen or carbon-nitrogen double bonds may be present, which may occur as part of keto-enol and imine-enamine tautomerisation respectively. Independent from its constitution, an alkyl group, as defined herein, can be substituted or unsubstituted. Substituents include, but are not limited to, one or more alkyl groups, halogen atoms, haloalkyl groups, (un)substituted aryl groups, (un)substituted heteroaryl groups, amino, oxo, nitro, cyano, azido, hydroxy, mercapto, keto, carboxy, carbamoyl, expoxy, methoxy, ethynyl. In case alkyl, as defined herein, contains a poly ethylene glycol (PEG) moiety, the preferred number of carbon atoms can be exceeded by the number present in the PEG moiety, wherein the PEG moiety can contain a total amount of 1 to 500 ethylene glycol groups (—(CH₂CH₂O)_n— with n=1 to 500).
It has to be noted, that -(EO)_n— is used as an abbreviated expression for —(CH₂CH₂O)_n— with n indicating the number of ethylene glycol groups. The number of ethylene glycol groups especially may be n=1-500 or as stated in the particular example.
Aralkyl refers to an alkyl group as described above, that connects to an unsubstituted or substituted aromatic or heteroaromatic hydrocarbon moiety, consisting of one or more aromatic or heteroaromatic rings with 3-8 ring atoms each. Substituents for both the alkyl and aryl part include, but are not limited to, one or more halogen atoms, alkyl or haloalkyl groups, substituted or unsubstituted aryl groups, substituted or unsubstituted heteroaryl groups, amino, nitro, cyano, hydroxy, mercapto, carboxy, azido, methoxy, methylthio.
Aryl refers to an aryl group, which is an unsubstituted or substituted aromatic or heteroaromatic hydrocarbon moiety, consisting of one or more aromatic or heteroaromatic rings with 3-8 ring atoms each. Substituents include, but are not limited to, one or more halogen atoms, haloalkyl groups, substituted or unsubstituted aryl groups, substituted or unsubstituted heteroaryl groups, amino, nitro, cyano, hydroxy, mercapto, carboxy, azido, methoxy, methylthio.
Acyl refers to a —C(O)-alkyl group, wherein the alkyl group is as defined above.
Aracyl refers to a —C(O)-aryl group, wherein the aryl group is as defined above.
Carbamoyl refers to a —C(O)—NH₂group, wherein the hydrogens can independently from each other be substituted with an alkyl group, aryl group or aralkyl group, wherein alkyl group, aryl group or aralkyl group are as defined above.
O-acyl refers to an —O—C(O)-alkyl group, wherein the alkyl group is as defined above.
O-alkyl refers to an alkyl group, which is bound through an O-linkage, wherein the alkyl group is as defined above.
O-aracyl refers to a —O—C(O)-aryl group, wherein the aryl group is as defined above.
O-aralkyl refers to an aralkyl group, which is bound through an O-linkage, wherein the aralkyl group is as defined above.
O-aryl refers to an aryl group, which is bound through an O-linkage, wherein the aryl group is as defined above.
O-carbamoyl refers to a carbamoyl group, which is bound through an O-linkage, wherein the carbamoyl group is as defined above.
S-alkyl refers to an alkyl group, which is bound through a S-linkage, wherein the alkyl group is as defined above.
S-aryl refers to an aryl group, which is bound through a S-linkage, wherein the aryl group is as defined above. S-aralkyl refers to an aralkyl group, which is bound through a S-linkage, wherein the aralkyl group is as defined above.
S-aralkyl refers to an aralkyl group, which is bound through an S-linkage, wherein the aralkyl group is as defined above.
Se-alkyl refers to an alkyl group, which is bound through a Se-linkage, wherein the alkyl group is as defined above. Se-aryl refers to an aryl group, which is bound through a Se-linkage, wherein the aryl group is as defined above.
Se-aralkyl refers to an aralkyl group, which is bound through a Se-linkage, wherein the aralkyl group is as defined above. NH-alkyl and N-bisalkyl refer to alkyl groups, which are bound through an N linkage, wherein the alkyl groups are as defined above.
NH-aryl and N-bisaryl refer to aryl groups, which are bound through an N linkage, wherein the aryl groups are as defined above.
NH-carbamoyl refers to a carbamoyl group, which is bound through an N-linkage, wherein the carbamoyl group is as defined above.
Amido-alkyl refers to an alkyl group, which is bound through a NH—C(O)— linkage, wherein the alkyl group is as defined above.
Amido-aryl refers to an aryl group, which is bound through a NH—C(O)— linkage, wherein the aryl group is as defined above.
Amido-aralkyl refers to an aralkyl group, which is bound through a NH—C(O)— linkage, wherein the aralkyl group is as defined above.
Endstanding group refers to a group of a particular residue (R₁, R₄and/or R₅) which is (sterically) accessible and capable for covalently binding to a particular linking residue (LR^1-4). This may be a group at the actual terminal end of the residue (R₁, R₄and/or R₅) or at any terminal end of any sidechain of the residue (R₁, R₄and/or R₅), or which is otherwise located in the residue (R₁, R₄and/or R₅) and sufficiently (sterically) accessible and capable for covalently binding to a particular linking residue (LR^1-4). The definition of the term endstanding group, if applicable, is independently also valid for the residues LR⁵and/or LR^PEG. Further, the term terminus refers to an endstanding group which is actually a terminal end of the concerned residue.
The person skilled in the art is well aware that a particular linking residue (LR^1-4) may represent a radical depending on the number of particular G units it binds to. Thus, in compounds of formula (II), the particular linking residue (LR^1-4) may be a biradical, or in case it is (intermediary) bound to only one particular G unit it may be a monoradical. Similarly, in case of compounds formula (I), depending on the number of particular G units it binds to, the particular linking residue (LR¹) may be a biradical, triradical, or tetraradical, or in case it is (intermediary) bound to only one particular G unit it may be a monoradical.
If an otherwise considered monovalent group is used with the modifier “divalent” as in “divalent alkyl” then this adds a second attachment point. Non limiting examples of divalent alkyl would be —CH₂—, —CH₂CH₂—, —CH₂C(CH₃)₂CH₂—.
Whenever side chains or residues are depicted as “floating groups” on a ring system, for example, in the formula:
then these side chains (or residues) may replace any hydrogen atom attached to any of the ring atoms, including depicted, implied, or expressly defined hydrogen, as long as a stable structure is formed. All resulting substitution patterns are thus included. For the given example, this corresponds to
The person skilled in the art understands that many compounds that fall under formula III as defined above have tautomeric forms. It has to be noted that according to this specification all tautomeric forms fall under formula III if at least one of the tautomers falls under formula III as defined above.
In the chair form of saturated six-membered rings, bonds to ring atoms, and the molecular entities attached to such bonds, are termed “axial” or “equatorial” according to whether they are located about the periphery of the ring (“equatorial”), or whether they are orientated above or below the approximate plane of the ring (“axial”). Due to the given stereochemistry of the cyclic phosphate ring, the axial position can only be above the approximate plane of the ring.
In naturally occurring cyclic nucleotide monophosphates (cNMP), both R7 and R8 are oxygen, and the phosphorus double bond is “distributed or dislocated” between both atoms. In water at physiological pH, the compound has a negative charge between both oxygens, and a corresponding cation, such as H+ or Na+. The corresponding compound structures herein are presented as charged compounds with a dislocated double bond at the phosphorus, as long as this is in accordance with valency rules. This style is chosen to account for, depict and disclose all possible “locations” of the phosphorous double bond and distribution of electron density or charge each within a single structure. The dislocated double bond, as used herein, depending on the nature of the particular R7 and R8, however, does not necessarily refer to an equally distributed charge or electron density between R7 and R8.
The term PLM as used herein, stands for polymer linked multimeric guanosine-3′, 5′-cyclic monophosphate (cGMP) analogue, wherein the term “multimeric” can refer to di-, tri- or tetrameric. To ease understanding of definitions and descriptions the term PLD (polymer linked dimeric cGMP analogue) is used as an equivalent to specifically refer to dimeric compounds.
The term polymer, as used herein, refers to a moiety consisting of at least two (equal) units or monomers.
If R7 and R8 are not equal, the phosphorus atom has four different ligands and becomes chiral resulting in two stereoisomeric forms. To describe the configuration of the chiral phosphorus, the Rp/Sp-nomenclature is used. Therein R/S follows the Cahn-Ingold-Prelog rules while “p” stands for phosphorus.
To give an example: if the equatorial residue R₈is oxygen (while axial R₇is sulphur), the corresponding cyclic guanosine-3′, 5′-monophosphorothioate compound (cGMPS-analogue) is Sp-configurated at phosphorus.
The person skilled in the art knows, that for the use in the field of the medicine especially as part of medicaments certainly only physiologically acceptable salts of the compounds according to the invention may be used.

Further Specification of Structures

In an embodiment the invention relates to a compound according to the definition hereinabove, wherein in case of formula (I) G⁴is absent, or, wherein in case of formula (II) G⁴and LR⁴are absent.
In another embodiment the invention relates to a compound according to the definition hereinabove, wherein in case of formula (I) G³and G⁴are absent, or, wherein in case of formula (II) G³, G⁴, LR³and LR⁴are absent.
In an embodiment the invention relates to a compound according to any definition hereinabove, wherein all R₇are O and all R₈are OH.
According to the invention it is preferred, that linking residues LR¹, LR², LR³and LR⁴are further subdivided as depicted in formula (Ib) and (IIb),
wherein:
coupling functions C¹, C^1′, C², C^2′, C³, C^3′, C⁴and C^4′ independently from each other can be absent or as defined by structures selected from the group consisting of
while connectivity can be as depicted or reversed as exemplified by

- G¹-O—C(O)—NH—S²versus G¹-NH—C(O)—O—S²

and wherein
in case the coupling function (C¹, C^1′, C², C^2′, C³, C^3′, C⁴and/or C^4′) does not replace the residue of the G unit (R₁, R₄and/or R₅of G^1-4) but bind to it, the particular residue (R₁, R₄and/or R₅) involved in coupling of G units (or G unit with dye(s) or other reporting group(s)) independently from each other is as defined further above, wherein an endstanding group is replaced by or transformed to a coupling function
or
selected from the group depicted hereinafter (wherein if present, Q₁connects to the G unit)




n = 0-6; m = 0-6;
Q₁= absent, S, NH, O, C(O),
S(O), S(O)₂;
Q₂= NH, S, O, C(O), CH₂,
OC(O), NC(O);



n = 0-4, m = 0-4
Q₁= absent, S, NH, O, C(O),
S(O), S(O)₂;



n₁= 0-4, n₂= 0-4, n₃= 0-4,
Q₁= absent, S, NH, O, C(O),
S(O), S(O)₂;
Q₂= NH, S, O, C(O), CH₂,
OC(O), NC(O);



n = 0-4; m = 0-4;
Q₁= absent, S, NH, O, C(O),
S(O), S(O)₂;



n1 = 0-4, n2 = 0-4; m = 0-4;
Q₁= absent, S, NH, O, C(O),
S(O), S(O)₂;
Q₂= CH₂, O, NH, S;

and wherein
the linker (L) is selected from the group consisting of


Dimeric Linkers	Trimeric Linkers



n = 0-4	n = 0-4



X = CH₂, O, NH, S, S—S, C(O);	X = CH₂, O, NH, S, S—S, C(O);
n = 0-4	n = 0-4



X = CH₂, NH, O, S;	X = CH₂, NH, O, S;
n = 0-4; m = 1-2	n = 0-4; m = 1-2



n = 0-4	n = 0-4


n = 0-4	n = 0-4



n = 0-4	n = 0-4



X = NH, O, S; n = 0-4;	X = NH, O, S; n = 0-4;



n = 0-6	n = 0-12



	n = 0-6



	n = 0-4



	X = O, S; n = 1-4



	X = CH, P; n = 0-4



	X = B, Si; n = 1-4



	n = 1-4 or



	n = 1-4



	Tetrameric linkers



	n = 0-4



	X = CH₂, O, NH, S, S—S, C(O);
	n = 0-4 or



	X = CH₂, O, NH, S, S—S, C(O);
	n = 0-4



	X = CH₂, NH, O, S, C(O); n = 0-4; m = 1-2



	n = 0-4 or



	n = 0-4



	n = 0-4



	X = NH, O, S; n = 0-4;



	X = C, Si; n = 0-6 or



	m = 0-24, n = 0-6



	n = 0-6



	n = 0-4



	X = O, S; n = 1-4



	n = 0-4



	n = 1-4



	n = 1-6; m = 1-11



	X = N, CH; n = 1-6; m = 0-10



	n = 0-6

while

- n for each sidechain within a particular linker of the list herebefore can have an equal or individual value as defined
- and
- all chiral, diastereomeric, racemic, epimeric, and all geometric isomeric forms of linkers (L) of the list herebefore, though not explicitly depicted, are included herein
- and
- cationic linkers (L) such as ammonium-derivatives are salts containing chloride-, bromide-, iodide- phosphate-, carbonate-, sulfate-, acetate- or any other physiologically accepted counterion

and wherein
spacers (S¹, S², S³and S⁴) can be equal or individual within a particular compound, be absent or be —(CH₂)_n1—(CH₂CH₂ß)_m-(CH₂)_n2— (with ß=O, S or NH; m=1 to 500, n1=0 to 8, n2=0 to 8, while both n1 and n2 can independently be equal or individual), or —(CH₂)_n— (with n=1 to 24).
Particularly, in the preferred embodiment of the invention, wherein it is preferred, that linking residues LR¹, LR², LR³and LR⁴are further subdivided as depicted in formula (Ib) and (IIb), containing spacer moieties (S^1-4), coupling functions (C^1-4, C^1-4) and a linker (L, only multimers of structure Ib), coupling functions (C^1-4, C^1-4) establish covalent bonds between

- the spacer and a G unit (G^1-4) by connecting to or replacing any of the residues R₁, R₄and/or R₅(compare formula structure III)
- and/or
- the spacer and a linker (L), dye or another reporting group
- and/or
- (in case the particular spacer is absent) a G unit (G^1-4) and a dye or another reporting group by connecting to or replacing any of the residues R₁, R₄and/or R₅
- and/or
- (in case the particular spacer is absent and/or a G unit is replaced by a dye or other reporting group) the linker (L) and a dye or another reporting group or a G unit (G^1-4, by connecting to or replacing any of the residues R₁, R₄and/or R₅).

Coupling functions (C^1-4, C^1′-4) are generated in a reaction between endstanding groups of the particular precursor parts according to well established methods of the art. Non limiting examples of precursor endstanding groups (of monomeric G units and (commercially available) linkers, dyes, reporting groups and spacers) and the corresponding coupling functions (C^1-4, C^1′-4), to which they are transformed within the assembled (mono- or multimeric) compound according to the invention, are as depicted in Table 1. Coupling functions (C^1-4, C^1′-4) can independently further be absent or be equal or individual within a particular mono- or multimeric compound.

TABLE 1

Endstanding groups and corresponding coupling functions (C^1-4, C^1′-4′)

		(Corresponding) Coupling
	Endstanding Group of	Function

	Monomeric G unit (R₁, R₄, R₅),	Spacer	(C¹, C^1′, C², C^2′, C³, C^3′, C⁴,
Entry	L, dye, reporting group	(S¹, S², S³, S⁴)	C^4′)

1

2

3

4

5

6

7

8

9



10

11

12

13

14

15

16

17

18

19

20

21

	X₁= Br, I	X₂= alkyl, OH, O-alkyl	X₃= NH, O, S
	bound to aryl, heteroaryl,	X₃= NH, O, S
	alkenyl

22

	X₁= Br, I	X₂= alkyl, OH, O-alkyl	X₃= NH, O, S
	bound to aryl, heteroaryl,	X₃= NH, O, S
	alkenyl


23

	X₁= Br, I	X₂= alkyl, OH, O-alkyl
	bound to aryl, heteroaryl,
	alkenyl

24

	X₁= Cl, Br, I, OTf
	bound to aryl, heteroaryl,
	alkenyl

25

26

27

A person skilled in the art understands, that synthetic equivalents of the precursor endstanding groups of Table 1, such as but not limited to NHS esters instead of carboxylic acids or triflates instead of halogens can be used as well to generate the particular corresponding coupling function. A person skilled in the art further understands, that endstanding groups of the synthetic precursors (residues R₁, R₄and/or R₅, linker (L), dye, reporting group and spacer (S^1-4)) can be interchanged amongst each other, resulting in reversed connectivity of the coupling function within the mono- or multimeric analogue.
A non limiting example of a multimeric compound according to the invention, illustrating the used and defined variables above is given in FIG. 2.

DETAILED DESCRIPTION OF THE INVENTION

Embodiments of the invention are directed to new polymer linked multimeric guanosine-3′, 5′-cyclic monophosphate (cGMP) analogues that modulate the cGMP-signaling system, preferably having activating properties, and more preferably being activators of cGMP dependent protein kinase (PKG), and related monomeric precursors thereof. The invention is also directed to related monomeric compounds, which may also show modulating activity and/or may serve as monomeric precursors of the multimers.
The idea of addressing more than one binding site of a target protein simultaneously with a single molecule has been reported once before using a polymer linked dimeric cGMP analogue (PLD).⁵Therein a homologous series of one PLD (FIG. 1), differing only in the length of the PEG spacer, was synthesized and tested for the ability to activate cGMP-dependent protein kinase Iα (PKG Iα) and cyclic nucleotide-gated ion channels (CNG channels). The results suggested, that PLDs feature an enhanced activatory potential compared to monomeric cGMP, while this enhancement, however, fundamentally depends on an optimum spacer length (between the cGMP units), which is unique for each addressed protein. Thus with increasing deviation from this optimum spacer length the effect was reported to decrease and eventually to disappear. In particular the only studied PLD gave best PKG Iα activation with a PEG-spacer of 282 Da. The same compound carrying a PEG-spacer of 2000 Da on the other hand displayed very strong activation of CNG channels but virtually no increased effect on PKG Iα (compared to monomeric cGMP). Accordingly, addressing both PKG Iα and CNG channels (or multiple targets in general), with the same PLD, appeared, if feasible at all, only possible with an intermediate spacer length at which the activation potential for both targets would be significantly decreased. Furthermore, the impact of PLDs or other multimeric cGMP analogues on PKG isoforms Iß and II had not been studied before. In a quest to discover multimeric cGMP analogues, that would give similar results on PKG IR and/or PKG II, optionally with additional PKG Iα and/or CNGC interaction, a corresponding PLD (with a PEG 2000 Da spacer) was synthesized as part of the invention (PET-cGMP-8-TMAmd-(PEG pd 2000)AmdMT-8-cGMP-PET, Table 13, compound 13). Said compound features an amide—instead of the reported sulfonyl coupling function as well as an additional R-phenyl-1, N²-etheno (PET) function at both nucleobases. Unexpectedly, the PKG Iß activation potential determined for this new PLD is the strongest so far observed 1735-fold activity of cGMP) while PKG II is rather poorly activated (2.5-fold activity of cGMP). Very surprisingly and contrary to previously reported results of the comparable sulfonyl coupled PLD⁵, where superior PKG Iα activation disappeared at this spacer length, this compound not only causes increased PKG Iα activation but the effect is also more than 4-fold greater (≥140-fold potential of cGMP) as determined for the best PLD agonist (PEG-spacer of 282 Da) described before.
PKG activation values of the compounds of the invention are expressed as multiple of the cGMP activation with the cGMP activation value set as 1 for each isozyme. It has to be noted, that the applied standard assay conditions only allowed to determine increased activation potencies of up to 140-fold for PKG Iα, 2832-fold for PKG Iß and 408-fold for PKG II, which is due to the employed enzyme concentration in the assays and the phenomenon that the isozymes were activity-titrated in some cases by the highly active compounds of the invention. The actual PKG activation potentials of these particular corns pounds of the invention appear to be significantly higher and are therefore expressed as ≥140-fold for PKG Iα, ≥2832-fold for PKG Iß and ≥408-fold for PKG II.
Very surprisingly, for the analogue of compound 13 lacking the PET moiety (cGMP-8-TMAmd-(PEG pd 2000)-AmdMT-8-cGMP, Table 13, compound 10) the relative enhancement of PKG isoform activity (compared to cGMP) is almost the other way around. Thus it features the so far strongest reported PKG II activation potential (243-fold activity of cGMP) but shows a far less pronounced enhancement on relative PKG Iα and Iß activation (37-fold and 69-fold activity of cGMP). Accordingly, nucleobase manipulation of PLDs and/or variation of the coupling function, which both has not been studied before, surprisingly, overrules the so far proposed target selectivity induced by spacer length.^{8 5}In particular, variations at Ra and/or R₅(such as the PET moiety) and/or exchange of the sulfonyl coupling function, which overlaps with modifications of the R₁moiety, have a very significant impact on PKG activation. This was especially surprising and not foreseeable, as the monomeric precursors of the two PLD compounds, 8-Br-PET-cGMP and 8-Br-cGMP, differing only in the PET moiety, at least with respect to their PKG Iα and II activatory potential are very alike.²Yet, as described above, when coupled to a PLD, their PKG activation profile differs significantly. Accordingly, the PKG isoform activation profile of the PLD so far cannot be predicted from the one determined for the corresponding monomers. However, as described above, structural modifications were found to be essential parameters to estimate and manipulate the increasing effects of PLDs on relative PKG isoform activation.
PLDs of the present invention feature a variety of different spacer lengths, as the results described above were also reproduced with homologues PLDs. For instance shorter spacers (19 and 8 ethylene glycol units (-(EO)₁₉— and -(EO)₈—), see Table 13, compounds 12, 5, 8, 3) gave similar results, wherein in several cases the PKG enzyme was titrated (FIG. 3). As mentioned above, a titrated enzyme corresponds to a value beyond the measurement limit, indicating a very strong activation potential. The substantial major effect of nucleobase manipulation, in particular R₄/R₅substitution, can again be observed by comparing for instance compounds 3 and 5 (FIG. 3). Therein compound 3 is identical to 5 but lacks the PET moiety. It gives a more than 51-fold decreased relative PKG Iß activation compared to compound 5, while relative PKG II activation in turn is increased by a factor of 30. Thus, the described enhancing effect of R₄/R₅transformations on relative PKG isoform activation is not restricted to a particular spacer length.
Furthermore, the present invention comprises PLDs containing standard coupling moieties other than amide groups, as the superior activity of new PLDs according to the invention is not limited to this particular type of coupling function. Dimers linked via a triazole group (e.g. compound 23 with ≥2832-fold activity of cGMP for PKG IR, also see FIG. 4) or featuring no additional coupling function besides the thio ether group directly connected to the nucleobase (e.g. compound 1 with 231-fold activity of cGMP for PKG II), as two random non limiting examples of the present invention, gave comparable results. Both of these optional coupling strategies as well as amide type of coupling in addition avoid regiochemical issues during synthesis, proofed to be beneficially robust and provided beneficially high yields. Furthermore, all tested PLDs linked via the R₁position at all tested spacer lengths, wherein Ra is absent and R₅is NH₂(according to formula III), were highly potent activators of PKG II.
Another structural aspect of PLDs according to the invention concerns the linkage position at which the two cGMP analogues are coupled to each other. The observed activity enhancement of PLDs is not restricted to linkage via the R₁position. It is still present, when linkage is varied along the G unit. Thus, as a non limiting example, PLDs coupled via the PET-moiety (at R₄+R₅), displayed a similarly increased PKG agonist potential as PET-substituted derivatives tethered via the R₁-position (compound 6 and 23, FIG. 4). Accordingly PLDs of the invention comprise a variety of possible linking positions as defined further above.
The nature of the spacer moiety is another motive that affects PLD induced PKG activation. PEG (spacer) units used on the PLD derivatives mentioned so far, can be replaced by or combined with other functionalities such as peptides or alkanes, included in the present invention. Alkanes in particular, however, are restricted in size, as solubility decreases significantly with growing alkyl spacer length. Still alkyl spacers with moderate size (compounds 16, 19 and 21 as non limiting examples of the present invention) are tolerated with respect to maintaining sufficient water solubility. In addition a PKG activation screening performed with compound 16 indicated that such compounds can show an activity increasing effect (PKG II activation approx. 22-fold higher than for cGMP). This effect is less pronounced than for derivatives linked via PEG-spacers but alkyl chain tethered PLDs benefit from increased lipophilicity, which should support better cell permeability. Hence, making them useful tools for biochemical assays and for pharmacological applications.
As stated above, previously established and supported by the work of the inventors linkage to a second cGMP analogue unit appears to be essential to obtain significantly enhanced PKG activation. Moreover, prior to this invention just one type of PLD (FIG. 1) had been studied and its increased activation of PKG (tested only for the Iα isoform) was proposed to be connected to a simultaneous binding to homologous sites at two different PKG subunits (reasoned by the rather short observed optimal spacer length).⁵It was thus not foreseeable, that mixed (heterogenous) PLDs of the invention, featuring two unequal G units (e.g. containing one PET-cGMP unit and one that lacks the PET moiety, e.g. compound 11) with different binding affinities would give a PKG (isoform) activation profile, that to a large extend resembles the characteristics of both G units in their corresponding homogenous PLDs (in the exemplary case compound 12 and 8, FIG. 5). Mixed PLDs of the invention that additionally contain mixed linking positions (e.g. PET-cGMP analogue unit linked via the R₄+R₅-PET-moiety and unit lacking the PET-moiety linked via the R₁-position, e.g. 22) behaved similarly (FIG. 5). The latter non limiting example of mixed PLDs compound 22, which can be seen as a hybrid of the homogenous PLDs 6 and 18, particularly demonstrates the implied potential of mixed PLDs. Therein, compound 18 is a strong activator of PKG 11 (381-fold activity of cGMP) while 6 shows virtually no increased effect on this isoform (1.3-fold activity of cGMP). Even though 6 or this type of G unit respectively therefore appears to be unable to contribute to PKG II activation, the mixed PLD hybrid 22 still is a very strong activator of PKG II (194-fold activity of cGMP). These surprising results indicate, that linkage to a second cGMP (analogue) is required to obtain strongly enhanced PKG activity, the second G unit, however, does not necessarily need to be of the same kind. As described, the second G unit can even be a significantly less effective activator of PKG (observed for the respective homogenous PLD) while the superior PKG activation of the first G unit (again observed for the respective homogenous PLD) is substantially preserved within the mixed PLD hybrid.
These unexpected findings reveal another new great potential of (mixed) PLDs. As stated above, established effector compounds often need to be derivatized for specific biochemical applications. For instance, introduction of a fluorescent dye, to enable intracellular localization by means of microscopic or spectroscopic techniques, is a very common strategy. In order to obtain representative results, ideally such transformations, meant to facilitate assay read out, should have no impact on the target activation profile. However, these structural manipulations of the original compound frequently do result in a significant shift of target affinity and specificity or even loss of activation potential. This is especially the case, when the particular functionality can only be introduced at a pharmacophoric group or when it inhibits or weakens binding to the target protein due to steric hindrance. For applications that benefit from the use of multiple target compounds, in turn, a change in (or extension of) the target activation profile obviously can also be desirable. Developing a multi target compound, though, sometimes can be just as difficult as producing a target specific one. This is the case, whenever a modification, needed to address one target, inhibits binding to the second. Mixed PLDs as disclosed within the present invention, provide an improved solution to both of these problems. Their advantage springs from the fact, that two cGMP units (instead of one for monomers) contribute to the overall PKG activation profile. As described above, even such modifications, that would give a completely different target affinity (observed for the monomer or the homogeneous PLD), do not erase the enhanced activation characteristics of the parent compound, as long as they are performed at only one cGMP unit. In this respect, the effect of structural manipulation at a single cGMP unit is buffered within mixed PLDs. Thus, mixed PLDs allow a much broader diversity of modifications (at one cGMP unit), while the undesired decrease of PKG activation, caused by these modifications, is much less pronounced if present at all. On the other hand, mixed PLDs also support the design of multi target compounds. Functional groups (e.g. PET-group), intended to address different targets (e.g. different PKG isoform) apparently can be installed at one cGMP unit, giving an extended target activation spectrum of the mixed PLD.
The present invention also comprises the extension of the described concept of polymer linked cGMP analogues from dimers to tri- and tetramers. Therein linkage of the particular G units is accomplished either in a linear or branched fashion (see formula I and II). Compounds 14 and 15 (Table 13) are two non limiting examples of the latter case, featuring particularly strong PKG II activation as predicted from analogues PLD derivatives lacking the PET-moiety (≥416-fold activity of cGMP for compound 14). The increased number of G units within tri- and tetramers results in even more diverse opportunities to combine (different) activator and target independent functionalized G units. While dimeric analogues only allow linkage of an activator G unit to either another one or a target independent functionalized G unit, for tri- and tetramers both can be provided within a single compound. Accordingly, compared to PLDs, they offer a broader scope of applications.
In summary the present invention has established the first activators of PKG Iß and PKG II with PLM structure, which are furthermore significantly improved when compared to state of the art compounds. Among the new PLM are also derivatives, which in addition activate PKG Iα, and mixed PLM, which amongst others are beneficial for functionalization and/or addressing all three PKG isoforms. Nucleobase modifications at R₄/R₅and R₁position as a key part of the invention, thereby proved to be powerful modifiers of PKG activation potential. These modifications were shown herein to be able to exceed and overrule the effect of varying spacer lengths, which before was suggested to be the main effector of target selectivity and activity increase (compared to the monomer).^5,8As an exemplary embodiment of the present invention, PLMs coupled via the R₁position, which overlaps with a nucleobase modification at R₁, wherein in addition R₄is absent and R₅is NH₂(according to formula III), were found to feature strongly increased PKG II activation potential (compared to the monomer). Prior art PLD compounds⁵(see FIG. 1) also fall under the scope of this general structural paradigm, however, only by coincidence, and are expressively disclaimed from the present invention. Their appearance in the art was connected to a different question, wherein a different target (PKG Iα and CNGC) was addressed and the crucial role of a different modifier (spacer length instead of nucleobase modification) was concluded. The new PLM compounds of the present invention, furthermore differ in and benefit from improved synthetic coupling strategies. Prior art synthetic protocol for PLDs involved coupling of a thiol-group in the 8-(R₁)position with a bifunctional PEG vinylsulfone.⁵The reported conditions, as published later and in accordance with our own experience, however, favour addition at the 7-(R₂) instead of the 8-(R₁)position. To replace the insufficient coupling strategy of prior art, various more robust, regioselective and higher yielding methods were developed for the present invention, involving for instance peptide (amid)- and click chemistry.
To test the effect of the new PKG activators of the invention in a cellular system, the 661W cell line was used and increase in cell death after treatment was assessed (for more details see examples section). The 661W cell line is a photoreceptor precursor cell line, which expresses PKG (FIG. 6). This makes them a suitable model for examining PKG activity using cell death as readout since increased PKG activity was previously associated with increased cell death.⁹Results were compared to untreated cells and to incubation with 8-Br-PET-cGMP as reference. 8-Br-PET-cGMP is a well established commercially available PKG activator, which has been applied in various cellular systems and is furthermore a synthetic precursor of some of the exemplary PLDs of the invention. All 12 exemplary tested PLDs led to increased cell death at one or more concentrations when compared to untreated cells as well as the reference compound 8-Br-PET-cGMP (FIG. 7). Therein the most potent PLDs of the invention display a 5-6 fold increase in cell death when compared to untreated cells and 3-4 fold increase in cell death when compared to the reference 8-Br-PET-cGMP.
Compounds that are selected based on effects on 661W cells are promising tools for research studies on the intracellular signaling pathways mediated by cGMP. Such studies should not be restricted to retinal cells but also include other cell lines sensitive to changes in cGMP levels or activating cGMP targets.¹⁰
The present invention as summarized above and defined in the claims, which are particularly incorporated into this description by reference in their entirety, shall be exemplified in the following in more detail by preferred embodiments.

Preferred Compounds According to the Invention

According the invention it is preferred that R₁is selected from group consisting of H, halogen, azido, nitro, alkyl, acyl, aryl, OH, O-alkyl, O-aryl, SH, S-alkyl, S-aryl, S-aralkyl, S(O)-alkyl, S(O)-aryl, S(O)aralkyl, S(O)-benzyl, S(O)₂-alkyl, S(O)₂-aryl, S(O)₂-aralkyl, amino, NH-alkyl, NH-aryl, NH-aralkyl, NR9R10, SiR13R14R15 wherein R9, R10, R13, R14, R15 are alkyl.
According to the invention it is further preferred that R₁is selected from the group consisting of H, CI, Br, I, F, N₃, NO₂, OH, SH, NH₂, CF₃, 2-furyl, 3-furyl, 2-bromo-5-furyl, (2-furyl)thio, (3-(2-methyl)furyl)thio, (3-furyl)thio, 2-thienyl, 3-thienyl, (5-(1-methyl)tetrazolyl)thio, 1,1,2-trifluoro-1-butenthio, (2-(4-phenyl)imidazolyl)thio, (2-benzothiazolyl)thio, (2,6-dichlorophenoxypropyl)thio, 2-(N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino)ethylthio, (4-bromo-2,3-dioxobutyl)thio, [2-[(fluoresceinylthioureido)amino]ethyl]thio, 2,3,5,6-tetrafluorophenylthio, (7-(4-methyl)coumarinyl)thio, (4-(7-methoxy)coumarinyl)thio, (2-naphtyl)thio, (2-(1-bromo)naphtyl)thio, benzimidazolyl-2-thiobenzothiazolylthio, 4-pyridyl, (4-pyridyl)thio, 2-pyridylthio, 5-amino-3-oxopentylamino, 8-amino-3,6-dioxaoctylamino, 19-amino-4,7,10,13,16-pentaoxanonadecylamino, 17-amino-9-aza-heptadecylamino, 4-(N-methylanthranoyl)aminobutylamino, dimethylamino, diethylamino, 4-morpholino, 1-piperidino, 1-piperazino, triphenyliminophosphoranyl or as depicted in Table 2.

TABLE 2

Residue R₁.

Entry	Residue

1





	wherein
	m = 0-6.
	Q = S, S(O), S(O)₂, O, NH, Se, CH₂, C(O).
	X₁, X₂and X₃can be equal or independently be H, OH, NH₂, N₃, SH, CN, NO₂, F, Cl, Br, I,
	(CH₂)_nCH₃(with n = 0-5), i-Pr, t-Bu, (CH₂)_nC≡CH (with n = 0-5), (CH₂)_nC═CH₂(with n = 0-5),
	CH₂OH, (CH₂)_nOCH₃(with n = 1-2), CH₂N(CH₃)₂, O(CH₂)_nCH₃(with n = 0-5), Oi-Pr, OCy,
	OCyp, OBn, OC(O)CH₃, OC(O)Ph, OCF₃, N(CH₃)₂, NH(CH₂)_nCH₃(with n = 0-5), NHC(O)t-Bu,
	NHC(O)Ph, NHC(O)Ot-Bu, NHC(O)CH₃, NHC(O)CH₂N₃, B(OH)₂, CF₃, C(O)OH, C(O)OCH₃,
	C(O)Oi-Pr, C(O)Ot-Bu, C(O)OPh, C(O)OBn, C(O)NH₂, C(O)N(CH₃)₂, C(O)NHPh, C(O)NHBn,
	C(O)CF₃, CH₂C(O)OH, CH₂C(O)OCH₃, CH₂C(O)Oi-Pr, CH₂C(O)Ot-Bu, CH₂C(O)OBn,
	S(CH₂)_nCH₃(with n = 0-5), S(CH₂)_nOEt (with n = 1-4), SBn, SO₂CH₃, SO₂CF₃,



	(with Y₁= H, SH, CN, Ph, F, CH₃, OCH₃, SCH₃, 4-thiophenyl, NO₂, pentyl),



	(with Y₂= H, SH, F),



	(with Y₃= H, SH),





2	or



	wherein
	m = 0-6.
	n = 1-6.
	Q = S, S(O), S(O)₂, O, NH, Se.

According to the invention it is especially preferred that R₁is selected from the group consisting of H, Cl, Br, I, F, N₃, NO₂, OH, SH, NH₂, CF₃, 2-furyl, 3-furyl, (2-furyl)thio, (3-(2-methyl)furyl)thio, (3-furyl)thio, 2-thienyl, 3-thienyl, (5-(1-methyl)tetrazolyl)thio, 1,1,2-trifluoro-1-butenthio, (2-(4-phenyl)imidazolyl)thio, (2-benzothiazolyl)thio, (2,6-dichlorophenoxypropyl)thio, 2-(N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino)ethylthio, (4-bromo-2,3-dioxobutyl)thio, [2-[(fluoresceinylthioureido)amino]ethyl]thio, 2,3,5,6-tetrafluorophenylthio, (7-(4-methyl)coumarinyl)thio, (4-(7-methoxy)coumarinyl)thio, (2-naphtyl)thio, (2-(1-bromo)naphtyl)thio, benzimidazolyl-2-thio, benzothiazolylthio, 4-pyridyl, (4-pyridyl)thio, 2-pyridylthio, 5-amino-3-oxopentylamino, 8-amino-3,6-dioxaoctylamino, 19-amino-4,7,10,13,16-pentaoxanonadecylamino, 17-amino-9-aza-heptadecylamino, 4-(N-methylanthranoyl)aminobutylamino, dimethylamino, diethylamino, 4-morpholino, 1-piperidino, 1-piperazino, triphenyliminophosphoranyl or as depicted in Table 3.

TABLE 3

Residue R₁.







wherein
m = 0-6.
Q = S, S(O), S(O)₂, NH.
X₁, X₂and X₃can be equal or independently be H, OH, NH₂, N₃, SH, CN, NO₂, F, Cl, Br, I, (CH₂)_nCH₃
(with n = 0-5), i-Pr, t-Bu, Ph, (CH₂)_nC≡CH (with n = 0-5), (CH₂)_nC═CH₂(with n = 0-5), CH₂OH,
(CH₂)_nOCH₃(with n = 1-2), CH₂N(CH₃)₂, O(CH₂)_nCH₃(with n = 0-5), Oi-Pr, OCy, OCyp, OPh, OBn,
OC(O)CH₃, OC(O)Ph, OCF₃, N(CH₃)₂, NH(CH₂)_nCH₃(with n = 0-5), NHC(O)t-Bu, NHC(O)Ph,
NHC(O)Ot-Bu, NHC(O)CH₃, NHC(O)CH₂N₃, B(OH)₂, CF₃, C(O)OH, C(O)OCH₃, C(O)Oi-Pr,
C(O)Ot-Bu, C(O)OPh, C(O)OBn, C(O)NH₂, C(O)N(CH₃)₂, C(O)NHPh, C(O)NHBn, C(O)CF₃,
CH₂C(O)OH, CH₂C(O)OCH₃, CH₂C(O)Oi-Pr, CH₂C(O)Ot-Bu, CH₂C(O)OBn, S(CH₂)_nCH₃(with
n = 0-5), S(CH₂)_nOEt (with n = 1-4), SBn, SPh,





or



wherein
m = 0-6.
n = 1-6.
Q = S, S(O), S(O)₂, NH.

According to the invention it is even more preferred that R₁is selected from the group consisting of H, Cl, Br, SH, 2-furyl, 3-furyl, (2-furyl)thio, (3-(2-methyl)furyl)thio, (3-furyl)thio, 2-thienyl, 3-thienyl, (5-(1-methyl)tetrazolyl)thio, 1,1,2-trifluoro-1-butenthio, (2-(4-phenyl)imidazolyl)thio, (2-benzothiazolyl)thio, (2,6-dichlorophenoxypropyl)thio, 2-(N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino)ethylthio, (4-bromo-2,3-dioxobutyl)thio, [2-[(fluoresceinylthioureido)amino]ethyl]thio, 2,3,5,6-tetrafluorophenylthio, (7-(4-methyl)coumarinyl)thio, (4-(7-methoxy)coumarinyl)thio, (2-naphtyl)thio, (2-(1-bromo)naphtyl)thio, benzimidazolyl-2-thio, benzothiazolylthio, 4-pyridyl, (4-pyridyl)thio, 2-pyridylthio, triphenyliminophosphoranyl or as depicted in Table 4.

TABLE 4

Residue R₁.







wherein
m = 0-3.
Q = S.
X₁, X₂and X₃can be equal or independently be H, OH, NH₂, N₃, SH, CN, NO₂, F, Cl, Br, I, (CH₂)_nCH₃
(with n = 0-5), i-Pr, t-Bu, Ph, (CH₂)_nC≡CH (with n = 0-5), (CH₂)_nC═CH₂(with n = 0-5), CH₂OH,
(CH₂)_nOCH₃(with n = 1-2), CH₂N(CH₃)₂, O(CH₂)_nCH₃(with n = 0-5), Oi-Pr, OCy, OCyp, OPh, OBn,
OC(O)CH₃, OC(O)Ph, OCF₃, N(CH₃)₂, NH(CH₂)_nCH₃(with n = 0-5), NHC(O)t-Bu, NHC(O)Ph,
NHC(O)Ot-Bu, NHC(O)CH₃, NHC(O)CH₂N₃, B(OH)₂, CF₃, C(O)OH, C(O)OCH₃, C(O)Oi-Pr,
C(O)Ot-Bu, C(O)OPh, C(O)OBn, C(O)NH₂, C(O)N(CH₃)₂, C(O)NHPh, C(O)NHBn, C(O)CF₃,
CH₂C(O)OH, CH₂C(O)OCH₃, CH₂C(O)Oi-Pr, CH₂C(O)Ot-Bu, CH₂C(O)OBn, S(CH₂)_nCH₃(with
n = 0-5), S(CH₂)_nOEt (with n = 1-4), SBn, SPh,





or



wherein
m = 0-6.
n = 1-6.
Q = S.

In addition to the above or independent to the above it is preferred that according the invention that R₄is selected from group consisting of H, amino, alkyl, aralkyl, nitro, N-oxide or R₄can form together with Y and R₅and the carbon bridging Y and R₅an imidazole ring which can be unsubstituted or substituted with alkyl, aryl or aralkyl, or can form together with Y and R₅and the carbon bridging Y and R₅an imidazolinone as depicted above (structure IV, V, n=1) or an homologous ring (n=2 to 8) which each can be unsubstituted or substituted (not depicted) with alkyl, aryl or aralkyl.
According to the invention it is further preferred that R₄is absent or selected from the group consisting of amino, N-oxide or as depicted in Table 5.

TABLE 5

Residue R₄.

Entry	Residue

1



	wherein
	m = 1-6.
	X₁, X₂and X₃can be equal or independently be H, N₃, CN, NO₂, F, Cl, Br, I, (CH₂)_nCH₃
	(with n = 0-5), i-Pr, t-Bu, (CH₂)_nC≡CH (with n = 0-5), (CH₂)_nC═CH₂(with n = 0-5), (CH₂)_nOCH₃
	(with n = 1-2), CH₂N(CH₃)₂, O(CH₂)_nCH₃(with n = 0-5), Oi-Pr, OBn, OC(O)Ph,
	OCF₃, N(CH₃)₂, NHC(O)t-Bu, NHC(O)Ph, NHC(O)Ot-Bu, NHC(O)CH₃, NHC(O)CH₂N₃, CF_3,
	C(O)OCH₃, C(O)Oi-Pr, C(O)Ot-Bu, C(O)OPh, C(O)OBn, C(O)NH₂, C(O)N(CH₃)₂, C(O)NHPh,
	C(O)NHBn, C(O)CF₃, CH₂C(O)OCH₃, CH₂C(O)Oi-Pr, CH₂C(O)Ot-Bu, CH₂C(O)OBn,
	S(CH₂)_nCH₃(with n = 0-5), S(CH₂)_nOEt (with n = 1-4), SBn, SPh, SO₂CF₃,



	(with Y₁= H, CN, Ph, F, CH₃, OCH₃, SCH₃, NO₂, pentyl),



	(with Y₂= H, F),



2



	wherein
	X₁can be H, CH₃, Ph.
	X₂can be H, Ph, 2-naphtyl, 9-phenanthryl, 1-pyrenyl, 2,3-dihydro-1,4-benzodioxin-6-yl,
	dibenzo[b,d]furan-2-yl, 2,3-dihydro-1-benzofuran-5-yl, 1-benzothien-5-yl, 1-benzofuran-5-yl,
	cycopropyl, 1-adamantyl, C(Ph)₃, 2-thienyl, 3-chloro-2-thienyl, 3-thienyl, 1,3-thiazol-2-yl, 2-
	pyridinyl, 5-chloro-2-thienyl, 1-benzofuran-2-yl,
	X₃, X₄and X₅can independently be H, OH, NH, CH₃, Cl, Br, F, CN, N₃, CF₃, OCF₃, NO₂,
	C(O)OH, C(O)OCH₃, OCH₃, OBn, O-benzoyl, SCH₃, t-Bu, N(CH₃)₂, S-phenyl, Ph, S(O)₂CH₃,
	C(O)NH₂, NHS(O)₂CH₃,

3



	wherein deviating from the definition above, any hydrogen atom attached to any of the ring
	carbon atoms including depicted, implied, or expressly defined hydrogen, or both hydrogen
	atoms (m = 2) attached to the same particular carbon atom, can be replaced by one or two
	(equal) “floating groups” X₁respectively, as long as a stable structure is formed.
	while m = 1 or 2.
	n = 1-4.
	X₁can be H, CH₃, Et, Pr, i-Pr, Bu, F, Ph, (CH₂)₂OH*
	*Only for first case.
4	or



	wherein
	m = 1-6.
	n = 1-6.

According to the invention it is especially preferred that R₄is absent or selected from the group consisting of amino, N-oxide or as depicted in Table 6.

TABLE 6

Residue R₄.

Entry	Residue

1



	wherein
	m = 1-3.
	X₁, X₂and X₃can be equal or independently be H, N₃, CN, NO₂, F, Cl, Br, I, (CH₂)_nCH₃
	(with n = 0-5), i-Pr, t-Bu, Ph, (CH₂)_nC≡CH (with n = 0-5), (CH₂)_nC═CH₂(with n = 0-5), (CH₂)_nOCH₃
	(with n = 1-2), CH₂N(CH₃)₂, O(CH₂)_nCH₃(with n = 0-5), Oi-Pr, OPh, OBn,
	OC(O)CH₃, OC(O)Ph, OCF₃, N(CH₃)₂, NHC(O)t-Bu, NHC(O)Ph, NHC(O)Oy-Bu,NHC(O)CH₃,
	NHC(O)CH₂N_3,CF_3,C(O)OCH₃, C(O)Oi-Pr, C(O)Ot-Bu, C(O)OPh, C(O)OBn, C(O)NH₂,
	C(O)N(CH₃)₂, C(O)NHPh, C(O)NHBn, C(O)CF₃, CH₂C(O)OCH₃, CH₂C(O)Oi-Pr,
	CH₂C(O)Ot-Bu, CH₂C(O)OBn, S(CH₂)_nCH₃(with n = 0-5), S(CH₂)_nOEt (with n = 1-4), SBn,
	SPh, SO₂CF₃,



2



	wherein
	X₁can be H, CH₃, Ph.
	X₂can be H, Ph, 2-naphtyl, 9-phenanthryl, 1-pyrenyl, 2,3-dihydro-1,4-benzodioxin-6-yl,
	dibenzo[b,d]furan-2-yl, 2,3-dihydro-1-benofuran-5-yl, 1-benzothien-5-yl, 1-benzofuran-5-yl,
	cycopropyl, 1-adamantyl, C(Ph)₃, 2-thienyl, 3-chloro-2-thienyl, 3-thienyl, 1,3-thiazol-2-yl, 2-
	pyridinyl, 1-benzofuran-2-yl;
	X₃, X₄and X₅can independently be H, OH, NH, CH₃, Cl, Br, F, CN, N₃, CF₃, OCF₃, NO₂,
	C(O)OH, C(O)OCH₃, OCH₃, OBn, O-benzoyl, SCH₃, t-Bu, N(CH₃)₂, S-phenyl, Ph, S(O)₂CH₃,
	C(O)NH₂, NHS(O)₂CH₃,

3	or


	wherein deviating from the definition above, any hydrogen atom attached to any of the ring
	carbon atoms including depicted, implied, or expressly defined hydrogen, or both hydrogen
	atoms (m = 2) attached to the same particular carbon atom, can be replaced by one or two
	(equal) “floating groups” X₁respectively, as long as a stable structure is formed.
	while m = 1 or 2.
	n = 1-4.
	X₁can be H, CH₃, Et, Pr, i-Pr, Bu, F, Ph, (CH₂)₂OH*
	*Only for first case.
4	or



	wherein
	m = 1-6.
	n = 1-6.

According to the invention it is even more preferred that R₄is absent or as depicted in Table 7.

TABLE 7

Residue R₄.

Entry	Residue

1



	wherein
	m = 1-3.
	X₁, X₂and X₃can be equal or independently be H, N₃, CN, NO₂, F, Cl, Br, I, (CH₂)_nCH₃
	(with n = 0-5), i-Pr, t-Bu, Ph, (CH₂)_nC≡CH (with n = 0-5), (CH₂)_nC═CH₂(with n = 0-5),
	(CH₂)_nOCH₃(with n = 1-2), CH₂N(CH₃)₂, O(CH₂)_nCH₃(with n = 0-5), Oi-Pr, OPh, OBn,
	OC(O)CH₃, OC(O)Ph, OCF₃, N(CH₃)₂, NHC(O)t-Bu, NHC(O)Ph, NHC(O)Ot-Bu, NHC(O)CH₃,
	NHC(O)CH₂N₃, CF_3,C(O)OCH₃, C(O)Oi-Pr, C(O)Ot-Bu, C(O)OPh, C(O)OBn, C(O)NH₂,
	C(O)N(CH₃)₂, C(O)NHPh, C(O)NHBn, C(O)CF₃, CH₂C(O)OCH₃, CH₂C(O)Oi-Pr,
	CH₂C(O)Ot-Bu, CH₂C(O)OBn, S(CH₂)_nCH₃(with n = 0-5), S(CH₂)_nOEt (with n = 1-4), SBn,
	SPh, SO₂CF₃,



2	or




	wherein
	X₁can be H, CH₃, Ph.
	X₂can be H, Ph, 2-naphtyl, 9-phenanthryl, 1-pyrenyl, 2,3-dihydro-1,4-benzodioxin-6-yl,
	dibenzo[b,d]furan-2-yl, 2,3-dihydro-1-benofuran-5-yl, 1-benzothien-5-yl, 1-benzofuran-5-yl,
	cycopropyl, 1-adamantyl, C(Ph)₃, 2-thienyl, 3-chloro-2-thienyl, 3-thienyl, 1,3-tiazol-2-yl, 2-
	pyridinyl, 1-benzofuran-2-yl;
	X₃, X₄and X₅can independently be H, OH, NH, CH₃, Cl, Br, F, CN, N₃, CF₃, OCF₃, NO₂,
	C(O)OH, C(O)OCH₃, OCH₃, OBn, O-benzoyl, SCH₃, t-Bu, N(CH₃)₂, S-phenyl, Ph, S(O)₂CH₃,
	C(O)NH₂, NHS(O)₂CH₃,
3	or



	wherein
	n = 1-4.
4	or



	wherein
	m = 1-3.
	n = 1-6.

In addition to the above or independent to the above it is preferred that according the invention that R₅is selected from the group consisting of H, halogen, NH-carbamoyl-alkyl, NH-carbamoyl-aryl, NH-carbamoyl-aralkyl, amino, NH-alkyl, NH-aryl, NH-aralkyl, NR₃OR₃₁wherein R₃₀and R₃₁are alkyl, or can form together with R₄, Y and the carbon bridging Y and R₅an imidazole ring which can be unsubstituted or substituted with alkyl, aryl or aralkyl, or can form together with R₄, Y and the carbon bridging Y and R₅an imidazolinone ring as depicted above (structure IV, V, n=1) or an homologous ring (n=2 to 8) which each can be unsubstituted or substituted (not depicted) with alkyl, aryl or aralkyl.
According to the invention it is further preferred that R₅is selected from the group consisting of H, NH₂, F, Cl, Br, I, methylamino, NH-benzyl, NH-phenyl, NH-4-azidophenyl, NH-phenylethyl, NH-phenylpropyl, 2-aminoethylamino, n-hexylamino, 6-amino-n-hexylamino, 8-amino-3,6-dioxaoctylamino, dimethylamino, 1-piperidino, 1-piperazino or can form together with R₄, Y and the carbon bridging Y and R₅a ring system as depicted in Table 5 (entry 2 and 3).
According to the invention it is especially preferred that R₅is selected from the group consisting of H, NH₂, F, Cl, Br, I, methylamino, NH-benzyl, NH-phenyl, NH-4-azidophenyl, NH-phenylethyl, NH-phenylpropyl, 2-aminoethylamino, n-hexylamino, 6-amino-n-hexylamino, 8-amino-3,6-dioxaoctylamino, dimethylamino, 1-piperidino, 1-piperazino or can form together with R₄, Y and the carbon bridging Y and R₅a ring system as depicted in Table 6 (entry 2 and 3).
According to the invention it is even more preferred that R₅is NH₂, or can form together with R₄, Y and the carbon bridging Y and R₅a ring system as depicted in Table 7 (entry 2 and 3).
In addition to the above or independent to the above it is preferred that according the invention that R₇is selected from group consisting of OH, O-alkyl, O-aryl, O-aralkyl, O-acyl, SH, S-alkyl, S-aryl, S-aralkyl, borano (BH₃), methylborano, dimethylborano, cyanoborano (BH₂CN), S-PAP, O-PAP, S-BAP, or O-BAP

- wherein PAP is a photo-activatable protecting group with PAP=o-nitro-benzyl, 1-(o-nitrophenyl)-ethylidene, 4,5-dimethoxy-2-nitro-benzyl, 7-dimethylamino-coumarin-4-yl (DMACM-caged), 7-diethylamino-coumarin-4-yl (DEACM-caged) and 6,7-bis(carboxymethoxy)coumarin-4-yl)methyl (BCMCM-caged).
- and wherein BAP is a bio-activatable protecting group with BAP=methyl, acetoxymethyl, pivaloyloxymethyl, methoxymethyl, propionyloxymethyl, butyryloxymethyl, cyanoethyl, phenyl, benzyl, 4-acetoxybenzyl, 4-pivaloyloxybenzyl, 4-isobutyryloxybenzyl, 4-octanoyloxybenzyl, 4-benzoyloxybenzyl.

According to the invention it is further preferred that R₇is selected from the group consisting of OH, methyloxy, ethyloxy, cyanoethyloxy, acetoxymethyloxy, pivaloyloxymethyloxy, methoxymethyloxy, propionyloxymethyloxy, butyryloxymethyloxy, acetoxyethyloxy, acetoxybutyloxy, acetoxyisobutyloxy, phenyloxy, benzyloxy, 4-acetoxybenzyloxy, 4-pivaloyloxybenzyloxy, 4-isobutyryloxybenzyloxy, 4-octanoyloxybenzyloxy, 4-benzoyloxybenzyloxy, acetyloxy, propionyloxy, benzoyloxy, SH, methylthio, acetoxymethylthio, pivaloyloxymethylthio, methoxymethylthio, propionyloxymethylthio, butyryloxymethylthio, cyanoethylthio, phenylthio, benzylthio, 4-acetoxybenzylthio, 4-pivaloyloxybenzylthio, 4-isobutyryloxybenzylthio, 4-octanoyloxybenzylthio, 4-benzoyloxybenzylthio, borano (BH₃), methylborano, dimethylborano, cyanoborano (BH₂CN).
According to the invention it is further more preferred that R₇is selected from the group consisting of OH, methyloxy, ethyloxy, cyanoethyloxy, acetoxymethyloxy, pivaloyloxymethyloxy, methoxymethyloxy, propionyloxymethyloxy, butyryloxymethyloxy, acetoxyethyloxy, acetoxybutyloxy, acetoxyisobutyloxy, phenyloxy, benzyloxy, 4-acetoxybenzyloxy, 4-pivaloyloxybenzyloxy, 4-isobutyryloxybenzyloxy, 4-octanoyloxybenzyloxy, 4-benzoyloxybenzyloxy, acetyloxy, propionyloxy, benzoyloxy, SH, methylthio, acetoxymethylthio, pivaloyloxymethylthio, methoxymethylthio, propionyloxymethylthio, butyryloxymethylthio, cyanoethylthio, phenylthio, benzylthio, 4-acetoxybenzylthio, 4-pivaloyloxybenzylthio, 4-isobutyryloxybenzylthio, 4-octanoyloxybenzylthio, 4-benzoyloxybenzylthio.
According to the invention it is even more preferred that R7 is OH.
In addition to the above or independent to the above it is preferred that according the invention that R8 is selected from group consisting of OH, O-alkyl, O-aryl, O-aralkyl, O-acyl, O-PAP or O-BAP,

- wherein PAP is a photo-activatable protecting group with non limiting examples of, optionally, PAP=o-nitro-benzyl, 1-(o-nitrophenyl)-ethylidene, 4,5-dimethoxy-2-nitrobenzyl, 7-dimethylamino-coumarin-4-yl (DMACM-caged), 7-diethylamino-coumarin-4-yl (DEACM-caged) and 6,7-bis(carboxymethoxy)coumarin-4-yl)methyl (BCMCM-caged); and wherein BAP is a bio-activatable protecting group with non limiting examples of, optionally, BAP=methyl, acetoxymethyl, pivaloyloxymethyl, methoxymethyl, propionyloxymethyl, butyryloxymethyl, cyanoethyl, phenyl, benzyl, 4-acetoxybenzyl, 4-pivaloyloxybenzyl, 4-isobutyryloxybenzyl, 4-octanoyloxybenzyl, 4-benzoyloxybenzyl;

According to the invention it is further preferred that R8 is selected from the group consisting OH, methyloxy, ethyloxy, cyanoethyloxy, acetoxymethyloxy, pivaloyloxymethyloxy, methoxymethyloxy, propionyloxymethyloxy, butyryloxymethyloxy, acetoxyethyloxy, acetoxybutyloxy, acetoxyisobutyloxy, phenyloxy, benzyloxy, 4-acetoxybenzyloxy, 4-pivaloyloxybenzyloxy, 4-isobutyryloxybenzyloxy, 4-octanoyloxybenzyloxy, 4-benzoyloxybenzyloxy, acetyloxy, propionyloxy, benzoyloxy;
According to the invention it is further even more preferred that R8 is OH.
In addition to the above or independent to the above it is preferred that according the invention residues involved in connecting a G unit with another G unit or a dye or another reporting group can be R₁, R₄and/or R₅, in which case the particular residue is

- as defined for the preferred embodiment above, wherein an endstanding group is replaced by or transformed to a coupling function
- or
- selected from the group depicted in Table 8 (wherein if present, Q1 connects to the G unit)

TABLE 8

Residues R1, R4 and R5 involved in connecting a G unit with another
G unit or a dye or another reporting group (if present
Q₁connects to the G unit).



n = 0-6; m = 0-6;
Q1 = absent, S, NH, O;
Q2 = NH, S, O, C(O), CH₂,
OC(O), NC(O);



n = 0-4, m = 0-4
Q1 = absent, S, NH, O;



n1 = 0-4, n = 0-4, n3 = 0-4,
Q1 = absent, S, NH, O;
Q2 = NH, S, O, C(O), CH₂,
OC(O), NC(O);

According to the invention it is further preferred that residues involved in connecting a G unit with another G unit or a dye or another reporting group can be R₁, R₄and/or R₅, in which case the particular residue is

- as defined for its preferred ordinary embodiment, wherein an endstanding group is replaced by or transformed to the coupling function
- or
- selected from the group depicted in Table 9 (wherein if present, Q1 connects to the G unit)

TABLE 9

Residues R₁, R₄and R₅involved in connecting a G unit with another G unit or a dye or
another reporting group (if present Q₁connects to the G unit)

R₁	R₄	R₄+ R₅







n = 0-6; m = 0-6;
Q1 = S;
Q2 = NH, S, O, CH₂, NC(O);

In addition to the above or independent to the above it is preferred that according the invention that coupling functions (C^1-4and C^1′-4) are absent or selected from the group depicted in Table 10.

TABLE 10

Coupling functions (C^1-4and C^1′-4′).















X₃= NH, O, S











X = O, S, NH, S(O), S(O)₂,
CH₂, C(O)

According to the invention it is further preferred that coupling functions (C^1-4and C^1′-4) are absent or selected from the group depicted in Table 11.

TABLE 11

Coupling functions (C^1-4and C^1′-4′).









X = O, S, NH, CH₂











X₃= NH, O, S

In addition to the above or independent to the above it is preferred that according the invention the linker (L) is absent or selected from the group depicted in Table 12.

TABLE 12

Linker (L).



n = 0-4



n = 0-6



n = 0-4



n = 0-12



n = 0-6



X = O, S; n = 1-4



X = B, Si; n = 1-4



X = N, CH; n = 1-6; m = 0-10



n = 0-4



X = C, Si; n = 0-6
or



m = 0-24, n = 0-6



n = 0-6



X = O, S; n = 1-4



n = 1-4



n = 1-6; m = 1-11

While n for each sidechain within a particular linker can have an equal or individual value as defined.
In addition to the above or independent to the above it is preferred that in case of formula (I) according to the invention G⁴or G⁴and G³are absent

- or in case of formula (II) G⁴and LR⁴or G⁴, LR⁴, G³and LR³are absent.

According to the invention it is even more preferred that in case of formula (I) G⁴and G³are absent

- or in case of formula (II) G⁴, LR⁴, G³and LR³are absent.

Particularly preferred embodiments of the invention based on the above exemplifications, are as defined in anyone of the claims 6, 7, 8 and 9.
Especially preferred according to the invention are the compounds of Table 13, and as defined in claim 10. It has to be noted that in case of doubt the chemical structure as depicted in the formula is the valid one. It further has to be noted, that the compounds of Table 13 are displayed as the free acid. The present invention, however, also comprises salts of these compounds, featuring cations such as but not limited to Na⁺, Li⁺, NH₄ ⁺, Et₃NH⁺ and (i-Pr)₂EtNH⁺.

TABLE 13

Structures of novel compounds according to the invention.

#	Compound	Structure

1	Guanosine-3′,5'-cyclic monophosphate-[8-thio- (pentaethoxy)-ethylthio-8]- guanosine-3′,5′-cyclic monophosphate

2	Guanosine-3′,5′-cyclic monophosphate-[8- thiomethylamidomethyl- (pentaethoxy)- propylamidomethylthio-8]- guanosine-3′,5′-cyclic monophosphate

3	Guanosine-3′,5′-cyclic monophosphate-[8- thiomethylamido-(octaethoxy)- ethylamidomethylthio-8]- guanosine-3′,5′-cyclic monophosphate

4	Guanosine-3′,5′-cyclic monophosphate-[8-(4- thiophenylthio)-(pentaethoxy)- ethyl-(4-thiophenylthio)-8]- guanosine-3′,5′-cyclic monophosphate

5	β-Phenyl-1, N²-ethenoguanosine- 3′,5′-cyclic monophosphate-[8- thiomethylamido-(octaethoxy)- ethylamidomethylthio-8]-β-phenyl- 1, N²-ethenoguanosine-3′,5′-cyclic monophosphate

6	8-Bromoguanosine-3′,5′-cyclic monophosphate-[1, N²-etheno-β- phenyl-4-yl-(1-[1,2,3]-triazole- 4-yl)-methoxy-(hexaethoxy)- methyl-(4-(4-[1,2,3]-triazole-1- yl)-β-phenyl-1, N²-etheno)]-8- bromoguanosine-3′,5′-cyclic monophosphate

7	Guanosine-3′,5′-cyclic monophosphate-[8- thiomethylamido-(octaethoxy)- ethylamidomethylthio-8]-β-phenyl- 1, N²-ethenoguanosine-3′,5′-cyclic monophosphate

8	Guanosine-3′,5′-cyclic monophosphate-[8- thiomethylamido- (nonadecaethoxy)- ethylamidomethylthio-8]- guanosine-3′,5′-cyclic monophosphate

9	Guanosine-3′,5′-cyclic monophosphate-[8-(1-[1,2,3]- triazole-4-yl)-methoxy- (hexaethoxy)-methyl-(4-[1,2,3]- triazole-1-yl)-8]-guanosine-3′,5′- cyclic monophosphate

10	Guanosine-3′,5′-cyclic monophosphate-[8- thiomethylamido-(PEG pd 2000)- amidomethylthio-8]-guanosine-3′, 5′-cyclic monophosphate

11	β-Phenyl-1, N²-ethenoguanosine- 3′,5′-cyclic monophosphate-[8- thiomethylamido- (nonadecaethoxy)- ethylamidomethylthio-8]- guanosine-3′,5′-cyclic monophosphate

12	β-Phenyl-1, N²-ethenoguanosine- 3′,5′-cyclic monophosphate-[8- thiomethylamido- (nonadecaethoxy)- ethylamidomethylthio-8]-β-phenyl- 1, N²-ethenoguanosine-3′,5′-cyclic monophosphate

13	β-Phenyl-1, N²-ethenoguanosine- 3′,5′-cyclic monophosphate-[8- thiomethylamido-(PEG pd 2000)- amidomethylthio-8]-β-phenyl-1, N²-ethenoguanosine-3′,5′-cyclic monophosphate

14	Benzene-1,3,5-tri-[(8- amidomethyl-(pentaethoxy)- propylamidomethylthio)guanosine- 3′,5′-cyclic monophosphate]

15	Ethylene glycol-bis(2- aminoethylether)-N,N,N',N'-tetra- [(8- methylamidoethylthio)guanosine-3′, 5′-cyclic monophosphate]

16	Guanosine-3′,5′-cyclic monophosphate-[8-thio- (dodecanyl)-thio-8]-guanosine-3′, 5′-cyclic monophosphate

17	β-Phenyl-1, N²-ethenoguanosine- 3′,5′-cyclic monophosphate-[8- thio-(dodecanyl)-thio-8]-β- phenyl-1, N²-ethenoguanosine-3′, 5′-cyclic monophosphate, triethyl ammonium salt

18	Guanosine-3′,5′-cyclic monophosphate-[8- thioethylamidomethyl-(1-[1,2,3]- triazole-4-yl)-methoxy- (hexaethoxy)-methyl-(4-[1,2,3]- triazole-1-yl)- methylamidoethylthio-8]- guanosine-3′,5′-cyclic monophosphate

19	Guanosine-3′,5′-cyclic monophosphate-[8-thioethylthio- 8]-guanosine-3′,5′-cyclic monophosphate

20	Guanosine-3′,5′-cyclic monophosphate-[8-thioethyl-(1- [1,2,3]-triazole-4-yl)-methoxy- (hexaethoxy)-methyl-(4-[1,2,3]- triazole-1-yl)-ethylthio-8]- guanosine-3′,5′-cyclic monophosphate

21	Guanosine-3′,5′-cyclic monophosphate-[8-thio- (dodecanyl)-(4-thiophenyl-4″- thiophenylthio)-(dodecanyl)-thio- 8]-guanosine-3′,5′-cyclic monophosphate

22	Guanosine-3′,5′-cyclic monophosphate-[8- thioethylamidomethyl-(1-[1,2,3]- triazole-4-yl)-methoxy- (hexaethoxy)-methyl-(4-(4-[1,2, 3]-triazole-1-yl)-β-phenyl-1, N²- etheno)]-8-bromoguanosine-3′,5′- cyclic monophosphate

23	β-Phenyl-1, N²-ethenoguanosine- 3′,5′-cyclic monophosphate-[8- thioethyl-(1-[1,2,3]-triazole-4- yl)-methoxy-(hexaethoxy)-methyl- (4-[1,2,3]-triazole-1-yl)- ethylthio-8]-β-phenyl-1, N²- ethenoguanosine-3′,5′-cyclic monophosphate

24	8-Bromoguanosine-3′,5′-cyclic monophosphate-[1- propylamidomethyl-(pentaethoxy)- propylamidomethylthio-8]-β- phenyl-1, N²-ethenoguanosine-3′, 5′-cyclic monophosphate

25	8-Bromoguanosine-3′,5′-cyclic monophosphate-[1-(pentaethoxy)- ethyl-1]-8-bromoguanosine-3′,5′- cyclic monophosphate

26	8-Bromoguanosine-3′,5′-cyclic monophosphate-[1- propylamidomethyl-(pentaethoxy)- propylamidopropyl-1]-8- bromoguanosine-3′,5′-cyclic monophosphate

27	8-Bromoguanosine-3′,5′-cyclic monophosphate-[1- propylamidomethyl-(pentaethoxy)- propylamidomethylthio-8]- guanosine-3′,5′-cyclic monophosphate

28	Guanosine-3′,5′-cyclic monophosphate-[8-(phenyl-4- thio)-(pentaethoxy)-ethyl-(4- thiophenyl)-8]-guanosine-3′,5′- cyclic monophosphate

29	β-1, N²-Acetyl-guanosine-3′,5′- cyclic monophosphate-[8- thiomethylamido-(octaethoxy)- ethylamidomethylthio-8]-β-1,N²- acetyl-guanosine-3′,5′-cyclic monophosphate

30	8-Phenylguanosine-3′,5′-cyclic monophosphate-[1, N²-etheno-(3- phenyl-4-yl-(1-[1,2,3]-triazole- 4-yl)-methoxy-(hexaethoxy)- methyl-(4-(4-[1,2,3]-triazole-1- yl)-β-phenyl-1, N²-etheno)]-8- phenylguanosine-3′,5′-cyclic monophosphate

Monomeric precursor cGMP analogues (G units) for the synthesis of polymer linked multimeric cGMP analogues (PLMs) are compounds of formula (III). As described above, the PKG activation potential is strongly increased, once the monomeric precursor is linked to additional one(s) within a PLM, wherein particularly enhanced PKG isoform activation can be related to a certain extend to structural parameters. Non limiting examples of methods for the transformation of monomeric precursors into exemplary PLMs are given in the examples section. In addition Table 1 gives an overview of exemplary endstanding groups, that can be used for coupling reactions and the corresponding coupling functions within the PLM, to which they are transformed according to established methods of the art.
The invention in one aspect also relates to monomeric compounds of formula (III) and/or monomeric precursors according to formula (III), of any compound of the invention as described herein above, wherein the monomeric compound of formula (III) and/or the monomeric precursor of formula (III) is defined in the context of any said compounds herein above, and preferably wherein the monomeric compound of formula (III) and/or monomeric precursor of formula (III) complies with the following proviso:

- R₇is O and R₅is OH

and further complies with at least one of the following provisos:

- R₄is not H and R₅is NH₂
  - wherein R₄is attached via a —CH₂— bridge, which is part of R₄
- or
- R₅together with R₄, Y and the carbon bridging Y and R₅form a ring system, which can be
  - a) an imidazolinone ring as depicted hereinafter (n=1) or an homologous ring (n=2 to 8)

- - b) an imidazole ring, which can be unsubstituted or substituted as depicted hereinafter as residue entry 1 and 2
    - residue entry 1

- - - wherein
    - X₁is H;
    - X₂can be H, 2-naphtyl, 9-phenanthryl, 1-pyrenyl, 2,3-dihydro-1,4-benzodioxin-6-yl, dibenzo[b,d]furan-2-yl, 2,3-dihydro-1-benzofuran-5-yl, 1-benzothien-5-yl, 1-benzofuran-5-yl, cyclopropyl, 1-adamantyl, C(Ph)₃, 2-thienyl, 3-chloro-2-thienyl, 3-thienyl, 1,3-thiazol-2-yl, 2-pyridinyl, 5-chloro-2-thienyl, 1-benzofuran-2-yl;
    - X₃, X₄and X₅can independently be OH, NH, CH₃, Cl, Br, F, CN, N₃, CF₃, OCF₃, NO₂, C(O)OH, C(O)OCH₃, OCH₃, OBn, O-benzoyl, SCH₃, t-Bu, N(CH₃)₂, S-phenyl, Ph, S(O)₂CH₃, C(O)NH₂, NHS(O)₂CH₃, while X₄and X₅can also independently be H;
    - residue entry 2

- - - while R₁is as in any of the compounds 31 to 107.
- or
- R₁is attached via a —S(O)— or —S(O)₂— bridge or via a carbon atom of an aromatic ring system, which in each case is part of R₁
  - while R₄is H and R₅is NH₂

and in addition complies with the proviso that the monomeric compound of formula (III) and/or the monomeric precursor compound of formula (III) is not selected from the group of compounds consisting of
and/or
the monomeric compound of formula (III) and/or the monomeric precursor of the invention is selected from the group depicted in Table 14.

TABLE 14

Structures of novel monomeric precursor compounds according to the invention.

#	Compound	Structure

31	8-Amidomethylthioguanosine- 3′,5′-cyclic monophosphate

32	8-(4-Boronatephenylthio)- guanosine-3′,5′-cyclic monophosphate

33	8-(4-Cyanobenzylthio) guanosine- 3′,5′-cyclic monophosphate

34	8-(4-(2-Cyanophenyl)- benzylthio)guanosine- 3′,5′-cyclic monophosphate

35	8-Cyclohexylmethyl- thioguanosine- 3′,5′-cyclic monophosphate

36	8-(2,4- Dichlorophenylthio) guanosine- 3′,5′-cyclic monophosphate

37	8-Diethylphos- phonoethylthio- guanosine-3′,5′-cyclic monophosphate

38	8-Ethylthioguanosine- 3′,5′-cyclic monophosphate

39	8-Hexylthioguanosine- 3′,5′-cyclic monophosphate

40	8-(4- Isopropylphenylthio) guanosine-3′,5′- cyclic monophosphate

41	8-(3-(2- Methyl)furanyl) thioguanosine-3′,5′- cyclic monophosphate

42	8-(5-(1- Methyl)tetrazolyl) thioguanosine- 3′,5′-cyclic monophosphate

43	8-(4- Methoxybenzylthio) guanosine- 3′,5′-cyclic monophosphate

44	8-(7-(4-Methyl) coumarinyl)thio- guanosine-3′,5′-cyclic monophosphate

45	8-Methylacetyl- thioguanosine-3′,5′- cyclic monophosphate

46	8-(5-(1- Phenyl)tetrazolyl) thioguanosine-3′,5′- cyclic monophosphate

47	8-(2-Phenylethyl) thioguanosine-3′,5′- cyclic monophosphate

48	8-(2-(4- Phenyl)imidazolyl) thioguanosine-3′,5′- cyclic monophosphate

49	8-(2-Thiophenyl) thioguanosine-3′,5′- cyclic monophosphate

50	8-(1,1,2-Trifluoro-1- butenthio)guanosine- 3′,5′-cyclic monophosphate

51	8-Amidopropyl- thioguanosine-3′,5′- cyclic monophosphate

52	8-Amidoethyl- thioguanosine-3′,5′- cyclic monophosphate

53	8-Amidobutyl- thioguanosine-3′,5′- cyclic monophosphate

54	8-Acetamidoethyl- thioguanosine-3′, 5′-cyclic monophosphate

55	8-(2-Benzothiazolyl) thioguanosine- 3′,5′-cyclic monophosphate

56	8-(2- Boronatebenzylthio) guanosine-3′,5′- cyclic monophosphate

57	8-(4-Boronatebutylthio) guanosine- 3′,5′-cyclic monophosphate

58	8-(4- Boronatebenzylthio) guanosine-3′,5′- cyclic monophosphate

59	8-(3- Boronatebenzylthio) guanosine-3′,5′- cyclic monophosphate

60	8-Azidomethyl- amidoethylthio- guanosine-3′,5′-cyclic monophosphate

61	8-(3- Boronatephenyl) amidobutylthio- guanosine-3′,5′-cyclic monophosphate

62	8-Benzylamidobutyl- thioguanosine- 3′,5′-cyclic monophosphate

63	8-Benzamidoethyl- thioguanosine- 3′,5′-cyclic monophosphate

64	8-(3-Boronatephenyl) amidomethyl- thioguanosine-3′,5′-cyclic monophosphate

65	8-Benzylamidomethylthio- guanosine-3′,5′-cyclic monophosphate

66	8-(3- Boronatephenyl) amidoethylthio- guanosine-3′,5′-cyclic monophosphate

67	8-(3- Boronatephenyl) amidopropylthio- guanosine-3′,5′-cyclic monophosphate

68	8-Carboxypropyl- thioguanosine-3′,5′- cyclic monophosphate

69	8-Carboxybutyl- thioguanosine-3′,5′- cyclic monophosphate

70	8-(2,6- Dichlorophenoxy- propyl)thio- guanosine-3′,5′-cyclic monophosphate

71	8-(4-Dimethylamino- phenyl)amido- methylthioguanosine- 3′,5′-cyclic monophosphate

72	8-(4-Dimethylamino- phenyl)amido- butylthioguanosine- 3′,5′-cyclic monophosphate

73	8-Ethylbutyrylthio- guanosine-3′,5′- cyclic monophosphate

74	8-Methylpropionyl- thioguanosine-3′,5′- cyclic monophosphate

75	8-Methylvalerianyl- thioguanosine- 3′,5′-cyclic monophosphate

76	8-Methoxyethyl- amidobutylthio- guanosine-3′,5′-cyclic monophosphate

77	8-Methoxyethyl- amidomethylthio- guanosine-3′,5′-cyclic monophosphate

78	8-Methoxyethyl- amidoethylthio- guanosine-3′,5′-cyclic monophosphate

79	8-Phenylamidomethylthio- guanosine-3′,5′-cyclic monophosphate

80	8-Phenylpropylthio- guanosine-3′,5′- cyclic monophosphate

81	8-(3-Butynylthio) guanosine-3′,5′- cyclic monophosphate

82	8-(4- Acetamidophenylthio) guanosine-3′,5′- cyclic monophosphate

83	8-(4- Chlorophenylsulfonyl) guanosine-3′,5′- cyclic monophosphate

84	8-(4-Chlorophenyl- sulfoxide)- guanosine-3′,5′-cyclic monophosphate

85	8-((2-Ethoxyethyl)-4- thiophenylthio) guanosine-3′,5′- cyclic monophosphat

86	8-(4-Thiophenyl-4″- thiophenylthio) guanosine-3′,5′- cyclic monophosphate

87	8-(2-Azidoethylthio) guanosine-3′,5′- cyclic monophosphate

88	8-(3-Aminopropyl)- (pentaethoxy)- methylamidomethylthio- guanosine- 3′,5′-cyclic monophosphate

89	8-(2-Aminoethyl)- (octaethoxy)- amidomethylthio- guanosine-3′,5′- cyclic monophosphate

90	8-(2-Bromoethyl)- (pentaethoxy)- (4-thiophenylthio) guanosine-3′,5′- cyclic monophosphate

91	8-(4-(Propargyloxy- (hexaethoxy)- methyl)-[1,2,3]- triazole-1-yl)- methylamidoethylthio- guanosine-3′, 5′-cyclic monophosphate

92	8-(4- Carboxyphenylthio) guanosine-3′,5′- cyclic monophosphate

93	8-(4-Hydroxy- phenylsulfonyl)- guanosine-3′,5′-cyclic monophosphate

94	8-(4-Isopropyl- phenylsulfonyl)- guanosine-3′,5′-cyclic monophosphate

95	8-(4-Methyl- carboxyphenylthio)- guanosine-3′,5′-cyclic monophosphate

96	8-Methylsulfonyl- guanosine-3′,5′- cyclic monophosphate

97	8-(1-Bromo-2- naphthyl)methyl- thioguanosine-3′,5′- cyclic monophosphate

98	8-(2-(1-Benzyl- [1,2,3]-triazole- 4-yl)-ethylthio) guanosine-3′,5′- cyclic monophosphate

99	8-(3-Fluoro-5- methoxybenzylthio) guanosine-3′,5′- cyclic monophosphate

100	8-Pentafluorobenzyl- thioguanosine- 3′,5′-cyclic monophosphate

101	8-Triphenylimino- phosphoranyl- guanosine-3′,5′-cyclic monophosphate

102	8-(4-Chlorophenyl) guanosine-3′,5′- cyclic monophosphate

103	8-(4-Fluorophenyl) guanosine-3′,5′- cyclic monophosphate

104	8-(2-Furyl)guanosine- 3′,5′-cyclic monophosphate

105	8-(4-Hydroxyphenyl) guanosine-3′,5′- cyclic monophosphate

106	8-(4-Isopropylphenyl) guanosine- 3′,5′-cyclic monophosphate

107	8-Phenylguanosine- 3′,5′-cyclic monophosphate

108	β-Phenyl-1, N²-etheno-8- thioguanosine-3′,5′-cyclic monophosphate

109	8-(2-Aminophenyl- thio)-ß-phenyl- 1, N²-ethenoguanosine-3′,5′- cyclic monophosphate

110	8-Cyclohexylthio- ß-phenyl-1, N²- ethenoguanosine- 3′,5′-cyclic monophosphate

111	8-Cyclopentylthio- ß-phenyl-1, N²- ethenoguanosine- 3′,5′-cyclic monophosphate

112	8-(4-Methylphenylthio)- ß-phenyl- 1, N²-ethenoguanosine- 3′,5′-cyclic monophosphate

113	8-(4-Methoxyphenylthio)-ß- phenyl-1, N²- ethenoguanosine-3′, 5′-cyclic monophosphate

114	8-(3-(2-Methyl) furanyl)thio-ß- phenyl-1, N²- ethenoguanosine-3′,5′- cyclic monophosphate

115	8-(7-(4-Methyl) coumarinyl)thio-ß- phenyl-1, N²- ethenoguanosine-3′,5′- cyclic monophosphate

116	8-(2-Naphthyl)thio- ß-phenyl-1, N²-ethenoguanosine- 3′,5′-cyclic monophosphate

117	ß-Phenyl-1, N²- etheno-8-(2- thiophenyl) thioguanosine-3′,5′- cyclic monophosphate

118	ß-Phenyl-1, N²- etheno-8-(2- phenylethyl) thioguanosine-3′,5′- cyclic monophosphate

119	8-Amidomethylthio- ß-phenyl-1, N²-ethenoguanosine- 3′,5′-cyclic monophosphate

120	8-Carboxymethylthio- β-phenyl-1, N²-ethenoguanosine- 3′,5′-cyclic monophosphate

121	8-(4-Boronate- phenylthio)-ß- phenyl-1, N²- ethenoguanosine-3′,5′- cyclic monophosphate

122	8-Ethylthio-ß-phenyl-1, N2- ethenoguanosine-3′,5′-cyclic monophosphate

123	8-(4-Fluorophenyl- thio)-ß-phenyl- 1, N²-ethenoguanosine- 3′,5′-cyclic monophosphate

124	8-Methylthio- ß-phenyl-1, N²- ethenoguanosine- 3′,5′-cyclic monophosphate

125	ß-Phenyl-1, N²-etheno-8- propylthio- guanosine-3′,5′-cyclic monophosphate

126	8-Azidoethylthio- β-phenyl-1, N²- ethenoguanosine- 3′,5′-cyclic monophosphate

127	ß-Phenyl-1, N²- etheno-8-(4- trifluoromethylphenyl- thio)guanosine- 3′,5′-cyclic monophosphate

128	8-(4-Chlorophenyl- sulfonyl)-β- phenyl-1, N²- ethenoguanosine-3′,5′- cyclic monophosphate

129	8-(4-Isopropyl- phenylthio)-β- phenyl-1, N²- ethenoguanosine-3′,5′- cyclic monophosphate

130	8-(4-Isopropyl- phenylsulfonyl)-β- phenyl-1, N²- ethenoguanosine-3′,5′- cyclic monophosphate

131	8-(4-Chlorophenyl)- β-phenyl-1, N²-ethenoguanosine- 3′,5′-cyclic monophosphate

132	8-(4-Hydroxyphenyl)- β-phenyl-1, N²-ethenoguanosine- 3′,5′-cyclic monophosphate

133	8-(4-Isopropylphenyl)- β-phenyl- 1, N²-ethenoguanosine- 3′,5′- cyclic monophosphate

134	8-Bromo-(4- methoxy-ß-phenyl- 1, N²-etheno) guanosine-3′,5′- cyclic monophosphate

135	8-Bromo-(4- methyl-ß-phenyl-1, N²-etheno)guanosine- 3′,5′-cyclic monophosphate

136	alpha-Benzoyl- beta-phenyl-1, N²-etheno-8- bromoguanosine-3′,5′- cyclic monophosphate

137	8-Bromo-(4-chloro- ß-phenyl-1, N²-etheno)guanosine- 3′,5′-cyclic monophosphate

138	8-Bromo-(3-nitro- ß-phenyl-1, N2-etheno)guanosine- 3′,5′-cyclic monophosphate

139	8-Bromo-(ß- tert.-butyl-1, N²- etheno)guanosine- 3′,5′-cyclic monophosphate

140	8-Bromo-(2- methoxy-ß-phenyl- 1, N²-etheno) guanosine-3′,5′- cyclic monophosphate

141	8-Bromo-(3- methoxy-ß-phenyl- 1, N²-etheno) guanosine-3′,5′- cyclic monophosphate

142	8-Bromo-(2,4- dimethoxy-ß- phenyl-1, N²- etheno)guanosine- 3′,5′-cyclic monophosphate

143	8-Bromo-(4- pyridinyl-1, N2- etheno)guanosine- 3′,5′-cyclic monophosphate

144	8-Bromo-(3- thiophen-yl-1, N2- etheno)guanosine- 3′,5′-cyclic monophosphate

145	8-Bromo-(4-fluoro- ß-phenyl-1, N2-etheno)guanosine- 3′,5′-cyclic monophosphate

146	8-Bromo-1, N2- ethenoguanosine- 3′,5′-cyclic monophosphate

147	8-Bromo-(3-hydroxy- ß-phenyl-1, N2-etheno)guanosine- 3′,5′-cyclic monophosphate

148	8-Bromo-(4- hydroxy-ß-phenyl-1, N²-etheno)guanosine- 3′,5′-cyclic monophosphate

149	8-Bromo-(ß-(2,3- dihydro-1,4- benzodioxin)-1, N2- etheno)guanosine- 3′,5′-cyclic monophosphate

150	8-Bromo-(4- methylsulfonamido- ß-phenyl-1, N2- etheno)guanosine- 3′,5′-cyclic monophosphate

151	8-Bromo-(4-cyano- β-phenyl-1, N²-etheno)guanosine- 3′,5′-cyclic monophosphate

152	8-Bromo-(α-phenyl- β-methyl-1, N²-etheno)guanosine- 3′,5′-cyclic monophosphate

153	β-(4-Aminophenyl)- 1, N²-etheno- 8-bromoguanosine- 3′,5′-cyclic monophosphate

154	8-Bromo-(6-methoxy- 2-naphthyl- 1, N²-etheno) guanosine-3′,5′- cyclic monophosphate

155	8-Bromo-(9- phenanthrenyl-1, N²- etheno)guanosine- 3′,5′-cyclic monophosphate

156	8-Bromo-(4- trifluoromethyl-β- phenyl-1, N²-etheno) guanosine-3′,5′- cyclic monophosphate

157	(4-Fluoro-ß- phenyl-1, N2- etheno)-8-methyl- thioguanosine-3′,5′- cyclic monophosphate

158	(4-Methoxy- ß-phenyl-1, N2- etheno)-8-methyl- thioguanosine-3′,5′- cyclic monophosphate

159	1, N²-Etheno-8-(2- phenylethyl) thioguanosine-3′,5′- cyclic monophosphate

160	(4-Methoxy-ß- phenyl-1, N2- etheno)-8-propylthio- guanosine-3′,5′- cyclic monophosphate

161	β-1, N²-Acetyl-8- bromoguanosine- 3′,5′-cyclic monophosphate

162	8-Bromo-δ-1, N²- butyrylguanosine- 3′,5′-cyclic monophosphate

163	8-Bromo-1-(3- carboxypropyl) guanosine-3′,5′- cyclic monophosphate

164	1-[Aminomethyl- (pentaethoxy)- propylamidopropyl]-8- bromoguanosine- 3′,5′-cyclic monophosphate

165	1-Benzyl-8-bromo- guanosine-3′,5′- cyclic monophosphate

166	2′-O-(2-Azidoacetyl)- 8-bromo-β- phenyl-1, N²- ethenoguanosine-3′,5′- cyclic monophosphate

As described above, the compounds according to the present invention may further be labelled, according to well-known labelling techniques. For example, fluorescent dyes may be coupled to the compounds in order to, but not limited to, localize the intracellular distribution of cyclic nucleotide binding proteins in living cells by means of confocal microscopy, for fluorescence correlation spectrometry, for fluorescence energy transfer studies, or for determination of their concentration in living cells.
It should be understood that hydrates of the compounds are also within the scope of the present invention.
Instead of or additional to fluorescent dyes the compounds according to the inventions may be labelled with (radio) nuclides. The person skilled in the art knows many techniques and suitable isotopes that can be used for this.
As described above, the invention also comprises PEGylated forms of the specified compounds, wherein PEGylation is generally known to greatly improve water solubility, pharmacokinetic and biodistribution properties.
The invention further comprises modifications wherein R₇(according to formula III) can be an unsubstituted or substituted thio- or borano function. Both modifications are known in the art to improve resistance towards metabolic degradation.^{1a, 14}
The invention also comprises prodrug forms of the described compounds, wherein the negative charge of the (modified or unmodified) phosphate moiety is masked by a bioactivatable protecting group. It is widely accepted that such structures increase lipophilicity and with that, membrane-permeability and bioavailability resulting in a 10-1000 fold enhanced potency compared to the mother-compound. Such bioactivatable protecting groups can be introduced according to well known techniques of the art and include, but are not limited to acetoxymethyl, propionyloxymethyl, butyryloxymethyl, pivaloyloxymethyl, acetoxyethyl, acetoxybutyl, acetoxyisobutyl. Non limiting examples of corresponding residues R7 and/or R8 according to the invention are acetoxymethyloxy, propionyloxymethyloxy and butyryloxymethyloxy. More labile examples of protecting groups include alkyl or aryl groups as well as substituted alkyl or aryl groups. Non limiting examples for chemically labile protection groups of the R7 and/or R8 position are methyl, ethyl, 2-cyanoethyl, propyl, benzyl, phenyl and polyethylene glycol. These compounds are inactive per se, but extremely membrane-permeable, leading to strongly increased intracellular concentrations. Upon hydrolysis of the ester bond, the biologically active mother compounds are released.
Compounds according to the invention can also feature a photolysable group (also-called “caged”- or photo-activatable protecting group), which can be introduced according to well known techniques of the art. For example, but not limited to, caged groups may be coupled to an R8 oxo-function, leading to compounds with significantly increased lipophilicity and bioavailability. Non limiting examples for caged groups are o-nitro-benzyl, 1-(o-nitrophenyl)-ethylidene, 4,5-dimethoxy-2-nitro-benzyl, 7-dimethylaminocoumarin-4-yl (DMACM-caged), 7-diethylamino-coumarin-4-yl (DEACM-caged) and 6,7-bis(carboxymethoxy)coumarin-4-yl)methyl (BCMCM-caged).
The compounds according to the present invention can also be immobilized to insoluble supports, such as, but not limited to, agarose, dextran, cellulose, starch and other carbohydrate-based polymers, to synthetic polymers such as polacrylamide, polyethyleneimine, polystyrol and similar materials, to apatite, glass, silica, gold, graphene, fullerenes, carboranes, titania, zirconia or alumina, to the surface of a chip suitable for connection with various ligands.
The compounds according to the present invention can also be encapsulated within nanoparticles or liposomes for directed or non-directed delivery and release purposes of the compounds as described in the literature.¹¹
Preferably, the new polymer linked multimeric cGMP analogues of the invention, or the related monomeric compounds of formula (III) of the present invention, respectively, are used for treating or preventing a disease or condition that is associated with low cGMP signaling activity.
Diseases and conditions are preferably treated with polymer linked multimeric cGMP analogues, or the related monomeric compounds of formula (III) of the present invention, respectively, that activate the disease-related unbalanced cGMP-system, and include^1c:

- neurodegenerative diseases associated with insufficient synaptic function and learning and memory defects.
- neuromuscular junction defects including motor neuron diseases (e.g. Amyotrophic lateral sclerosis (ALS), Primary lateral sclerosis), also forms caused by certain infectious diseases (e.g. paralytic Poliomyelitis)
- cancer, including the initiation of cancer cell apoptosis and the prevention of metastasis cardiovascular diseases, including hypertension, cardiac hypertrophy, angina pectoris, ischemia and stroke
- parasitic diseases caused by trypanosomes, including Malaria, Chagas, sleeping sickness
- borelliosis (lyme disease)
- pulmonary diseases and conditions, such as pulmonary fibrosis and pulmonary hypertension
- osteoporosis
- Autoimmune diseases associated with an excessive proliferation of B- and T-cells including but not limited to: multiple sclerosis, Crohn's disease, Hashimoto's disease, juvenile arthritis, myocarditis, and rheuma.

It is to be understood herein that the treatment of a pathology, condition or disorder also includes the prevention thereof, even if not explicitly mentioned, unless specifically otherwise indicated.
In another aspect, the invention relates to a method for treating or preventing any of the above pathologies, conditions or disorders by administration of a therapeutically or prophylactically effective amount of an equatorially modified cGMP-analogue of the invention to a subject in need of prophylaxis or therapy.
The compounds according to the present invention, including the related monomeric compounds of formula (III) of the present invention, respectively, can also be used as research tool compound, preferably as research tool compound in regard of a disease or disorder related to an unbalanced cGMP-system, preferably a disease or disorder selected from the group consisting of cancer, cardiovascular disease or disorder, or autoimmune disease or disorder, or neurodegenerative disease or disorder.
The invention is further illustrated by the following figures and examples describing preferred embodiments of the present invention which are, however, not intended to limit the invention in any way.

EXAMPLES

1. Compound Synthesis
General Experimental Methods
All applied solvents and reagents were available from commercial suppliers. 8-Br-cGMP, 8-Br-PET-cGMP and 4-N₃-PET-8-Br-cGMP were available from Biolog Life Science Institute (Bremen, Germany). 8-T-cGMP is established in the literature and was prepared analogously to PET-8-T-cGMP (see examples below). Solvents used were specified as analytical or hplc grade. Dimethyl sulfoxide was stored over activated molecular sieves for at least two weeks before use. Chromatographic operations were performed at ambient temperature. Both reaction progress and purity of isolated products were determined by reversed phase hplc (RP-18, ODS-A-YMC, 120-S-11, 250×4 mm, 1.5 mL/min), wherein UV detection was performed either at 263 nm, an intermediate wavelength suitable to detect most cyclic GMP products and—impurities, or at the λ_maxof the particular starting material or product. Syntheses were typically performed in a 20-200 μmol scale in 2 mL polypropylene reaction vials with screw cap (reactions requiring inert gas atmosphere and/or degassing were performed in round bottom flasks (typically 10 or 25 mL)). Dissolution of poorly soluble reactants was achieved through sonification or heating (70° C.) prior to addition of reagents. In case dissolution was not elicited by these techniques, which mainly applied to some cGMP analogues carrying a PET-moiety, the suspension was used. Purification of products was accomplished by preparative reversed phase hplc (RP-18, ODS-A-YMC, 12 nm-S-10, 250×16 mm, UV 254 nm). The eluent composition is described in the particular synthetic example and, unless stated otherwise, can be used for analytical purposes as well. Desalting of products was accomplished by repeatedly freeze-drying or by preparative reversed phase hplc (RP-18, ODS-A-YMC, 12 nm-S-10, 250×16 mm, UV 254 nm) according to standard procedures for nucleotides. Solutions were frozen at −70° C. for 15 min prior to evaporation, in case a speedvac concentrator was used to remove the solvent. Products were either isolated as sodium or triethylammonium salt, depending on the applied buffer. Yields refer to the fraction of isolated product featuring the reported purity. They were calculated from UV-absorbance at the λ_max, measured on a JASCO V-650 Spectrophotometer (JASCO Germany GmbH, Gross-Umstadt, Germany) according to Lambert-Beers law. Extinction coefficients were estimated from literature known values of structurally related compounds. Mass spectra were obtained with an Esquire LC 6000 spectrometer (Bruker Daltronics, Bremen, Germany) in the ESI-MS mode with 50% water/50% methanol as matrix.
Experimental Procedures for the Preparation of 8-Thio-Modified Guanosine-3′,5′-Cyclic Monophosphate Analogues
General Procedure A:
In a typical experiment the corresponding thiol reactant (8 eq) and NaOH (2 M, 4 eq) were added successively to a solution of the corresponding 8-Br-substituted cGMP analogue (sodium salt, 65 mM, 1 eq) in H₂O/i-PrOH (1:1, v/v). The reaction mixture was heated to 90° C. and stirred until the bromide starting material was completely consumed or no further reaction progress was observed. The solution was then allowed to reach room temperature, neutralized with HCl (1 M) and the solvent was removed through high vacuum evaporation with a speedvac concentrator. The residue was dissolved in water (1 mL) and washed with MTBE (3×).* The aqueous phase was evaporated under reduced pressure using a rotary evaporator, the residue redissolved in water, subjected to preparative reversed phase hplc and desalted, giving the 8-thio-modified cGMP analogue. *In case the residue was not soluble in water, the obtained suspension was washed with MTBE and (if necessary) diluted with MeOH to dissolve remaining precipitate.
General Procedure A2:
In a typical experiment the corresponding thiol(ate) reactant (4.5 eq) was added to a solution of the corresponding 8-Br-substituted cGMP analogue (sodium salt, 65 mM, 1 eq) in H₂O/i-PrOH (1:1, v/v). The reaction mixture was stirred at room temperature until the bromide starting material was completely consumed or no further reaction progress was observed. The solution was then adjusted to pH 6 with NaOH (10%) and the solvent was removed through high vacuum evaporation with a speedvac concentrator. The residue was dissolved in water (1 mL) and washed with CH₂Cl₂(3×). The aqueous phase was evaporated under reduced pressure using a rotary evaporator, the residue redissolved in water, subjected to preparative reversed phase hplc and desalted, giving the 8-thio-modified cGMP analogue.
General Procedure B:
In a typical experiment a solution of the 8-Br-substituted cGMP analogue (sodium salt, 87 mM, 1 eq) was added portionwise over 2 h to a suspension of the corresponding dithiol (50 mM in water/i-PrOH, 2:3, v/v, 10 eq) and NaOH (2 M, 5 eq). The reaction mixture was heated to 90° C. and stirred until the bromide starting material was completely consumed or no further reaction progress was observed. The solvent was removed through high vacuum evaporation with a speedvac concentrator. The residue was suspended in water (1 mL), neutralized with HCl (1 M) and filtered. The crude product solution was subjected to preparative reversed phase hplc and desalted, giving the thiol analogue.
General Procedure C:
In a typical experiment NaOH (2 M, 16 eq) and the corresponding thiol reactant (8 eq) were added successively to a solution of the 8-Br-substituted cGMP analogue (sodium salt, 200 mM, 1 eq) in borate buffer (100 mM, pH 12). The reaction mixture was heated to 90° C. and stirred until the bromide starting material was completely consumed or no further reaction progress was observed. The solution was then allowed to reach room temperature and neutralized with HCl (1 M). The solvent was removed under reduced pressure using a rotary evaporator. The residue was dissolved in water (1 mL), subjected to preparative reversed phase hplc and desalted.
General Procedure D:
In a typical experiment N,N-diisopropylethylamine (2 eq) and the corresponding bromide (1 eq) were added successively to a solution of the 8-SH-substituted cGMP analogue (sodium or triethylammonium salt, 100 mM, 1 eq) in DMSO. The reaction mixture was stirred until the thiol starting material was completely consumed or no further reaction progress was observed. The solvent was removed through high vacuum evaporation with a speedvac concentrator. The residue was dissolved in water (1 mL), washed with ethyl acetate (3×), subjected to preparative reversed phase hplc and desalted.
General Procedure E:
For the formation of dimeric cGMP analogues general Procedure D was followed using N,N-diisopropylethylamine (2 eq), the corresponding bis-bromide spacer (0.5 eq) and the 8-SH-substituted cGMP analogue (sodium or triethylammonium salt, 100 mM, 1 eq) in DMSO.
Experimental Procedure for the Transformation of Carboxylic Acid Ester Functionalized Guanosine-3′,5′-Cyclic Monophosphate Analogues into the Corresponding Carboxylic Acid or Amide
General Procedure F:
In a typical experiment NaOH (2 M, 10 eq) was added to a solution of the corresponding ester (80 mM, 1 eq) in water/MeOH (1:1, v/v). The reaction mixture was stirred until the ester starting material was completely consumed or no further reaction progress was observed. The solution was then neutralized with HCl (1 M) and the solvent was removed under reduced pressure using a rotary evaporator. The residue was dissolved in water (1 mL), subjected to preparative reversed phase hplc and desalted, giving the carboxylic acid analogue.
General Procedure G:
In a typical experiment the corresponding ester (1 eq) was dissolved in excess methanolic ammonia (4.2 M, 200 eq). The reaction mixture was stirred until the starting material was completely consumed or no further reaction progress was observed. The solvent was removed through high vacuum evaporation with a speedvac concentrator. The residue was dissolved in water (1 mL), neutralized with HCl (1 M) and filtered through a syringe filter. The crude product was subjected to preparative reversed phase hplc and desalted, giving the carboxylic acid amide analogue.
Experimental Procedures for the Formation of Amide Bonds with Guanosine-3′,5′-Cyclic Monophosphate Analogues
General Procedure H:
In a typical experiment HOBt (1.1 eq), N,N-diisopropylethylamine (2.2 eq) and EDC (1.1 eq) were added successively to a solution of the corresponding acid-substituted cGMP analogue (100 mM in DMSO, 1 eq) and the corresponding amine (1.1 eq)*. The reaction mixture was stirred until the starting material was completely consumed or no further reaction progress was observed. The solvent was removed through high vacuum evaporation with a speedvac concentrator. The residue was dissolved in water (1 mL) and washed with ethyl acetate (5×). The aqueous phase was evaporated under reduced pressure using a rotary evaporator, redissolved in water, subjected to preparative reversed phase hplc and desalted, giving the coupled cGMP analogue. *The less valuable reactant was added in slight excess, thus for the reaction with reversed functions the amine-substituted cGMP analogue (100 mM in DMSO, 1 eq) and the acid reactant (1.1 eq) were used.
General Procedure I:
In a typical experiment HOBt (1.1 eq), N,N-diisopropylethylamine (2.2 eq) and EDC (1.1 eq) were added successively to a solution of the corresponding acid-substituted cGMP analogue (100 mM in DMSO, 1 eq) and the corresponding bis-amino spacer (0.5 eq). Workup was performed as described in general procedure H, giving the dimeric cGMP analogue.
General Procedure J:
In a typical experiment N,N-diisopropylethylamine (2.2 eq) and PyBOP (1.1 eq) were added successively to a solution of the corresponding carboxylic acid-substituted cGMP analogue (100 mM in DMSO, 1 eq) and the corresponding amine (1.1 eq)*. The reaction mixture was stirred until the starting material was completely consumed or no further reaction progress was observed (usually <10 min). Water (100 μL) was added, stirring was continued for 10 min and the solvent was removed through high vacuum evaporation with a speedvac concentrator. The residue was dissolved in water (1 mL), if necessary the pH was adjusted to 6 with NaOH (2 M) or HCl (1 M) and the solution washed with ethyl acetate (5×). The aqueous phase was evaporated under reduced pressure using a rotary evaporator, redissolved in water, subjected to preparative reversed phase hplc and desalted, giving the coupled cGMP analogue. *The less valuable reactant was added in slight excess, thus for the reaction with reversed functions the amine-substituted cGMP analogue (100 mM in DMSO, 1 eq) and the acid reactant (1.1 eq) were used.
General Procedure K:
In a typical experiment a solution of the corresponding carboxylic acid-substituted cGMP analogue (100 mM in DMSO, 1 eq) was added portionwise over 40 min to a solution of the bis-amino spacer (400 mM in DMSO, 5 eq), N,N-diisopropyethylamine (2.2 eq) and PyBOP (1.1 eq). More PyBOP (1 eq) was added and the reaction mixture was stirred until the starting material was completely consumed or no further reaction progress was observed (usually <10 min). Workup was performed as described in general procedure J, giving the monomeric cGMP analogue coupling product.
General Procedure L:
General procedure J was followed using the corresponding acid-substituted cGMP analogue (100 mM in DMSO, 1 eq), the bis-amino spacer (0.5 eq), N,N-diisopropylethylamine (2.2 eq) and PyBOP (1.1 eq) to obtain the dimeric cGMP analogue.
General Procedure M:
General procedure J was followed using the corresponding amine-substituted cGMP analogue (33 mM in DMSO, 1 eq), the linker tri-acid (0.3 eq), N,N-diisopropylethylamine (2 eq) and PyBOP (1.3 eq) to obtain the trimeric cGMP analogue.
General Procedure N:
General procedure J was followed using the corresponding amine-substituted cGMP analogue (diisopropylethylammonium salt, 50 mM in DMSO, 1 eq)*, the linker tetra-acid (tetra-diisopropylethylammonium salt, 0.25 eq)*, N,N-diisopropylethylamine (3 eq) and PyBOP (1.3 eq) to obtain the tetrameric cGMP analogue. *To transform the reactants into the diisopropylethylammonium salt they were subjected to N,N-diisopropylethylamine (3 eq per acidic function) in water (0.1-0.3 M) and evaporated to dryness using a speedvac concentrator at high vacuum.
Experimental Procedures for the Preparation of 8-Sulfonyl- and 8-Sulfoxide-Modified Guanosine-3′,5′-Cyclic Monophosphate Analogues
General Procedure O:
In a typical experiment a solution of OXONE® (180 mM, 5 eq) in NaOAc buffer (2 M, pH 4.2) was added dropwise to a solution of the corresponding 8-thio-substituted cGMP analogue (40 mM, 1 eq) in water/MeOH (1:1, v/v). The reaction mixture was stirred until the thio starting material was completely consumed or no further reaction progress was observed. The solution was then neutralized with NaOH (2 M) and filtered through a syringe filter. The solvent was removed under reduced pressure using a rotary evaporator. The residue was dissolved in water (1 mL), subjected to preparative reversed phase hplc and desalted, giving the 8-sulfonyl-substituted cGMP analogue.
Derivatives featuring a modified phosphate function, sensitive to oxidation reactions, such as a phosphorothioate, were synthesized starting from the corresponding guanosine, while the (modified) phosphate group was then introduced according to well established methods of the art (e.g. thiophosphorylation protocol¹²) after oxidation of the 8-thio function.
General Procedure P:
General procedure O was followed, favoring the formation of the 8-sulfoxide-substituted cGMP analogue through shorter reaction time and decreased equivalents of oxidizing agent OXONE® (1.5 eq).
Experimental Procedure for the Generation of 8-Azidoalkylthio-Substituted Guanosine-3′,5′-Cyclic Monophosphate Analogues
General Procedure Q:
In a typical experiment NaN₃(22.5 eq) was added portionwise over 5 h to a solution of 1,2-dibromoalkane (1.5 M, 15 eq) in DMF in an amber flask. The reaction mixture was stirred for 23 h and the 8-SH-substituted cGMP analogue (triethylammonium salt, 1 eq) as well as N,N-diisopropylethylamine (1 eq) were added successively. Stirring was continued until the cGMP analogue starting material was completely consumed or no further reaction progress was observed (usually about 1 h). The solvent was removed through high vacuum evaporation with a speedvac concentrator. The residue was dissolved in water (1 mL) and washed with MTBE (5×). The aqueous phase was evaporated under reduced pressure using a rotary evaporator, the residue was redissolved in water, subjected to preparative reversed phase hplc and desalted, giving the 8-azidoalkylthio-substituted analogue.
Experimental Procedures for the [3+2] Cycloaddition of Azides and Terminal Alkynes on Guanosine-3′,5′-Cyclic Monophosphate Analogues
General Procedure R:
In a typical experiment a solution of the corresponding azide (0.5 M in CH₂Cl₂, 1.1 eq) was added to the alkyne-substituted cGMP analogue (40 mM in H₂O, 1 eq) in an amber flask. Bromotris(triphenylphosphine)copper(I) ([Cu(PPh₃)₃Br]) (0.05 eq) was added and the reaction mixture was stirred until the alkyne starting material was completely consumed or no further reaction progress was observed. The mixture was diluted with water (to 1.5 mL) and washed with CH₂Cl₂(3×). The aqueous phase was evaporated under reduced pressure using a rotary evaporator, the residue was redissolved in water, subjected to preparative reversed phase hplc and desalted, giving the triazole addition product.
General Procedure S:
In a typical experiment Cu(PPh₃)₃Br (0.05 eq) was added to a solution of the corresponding azide (13 mM, 1 eq) and the corresponding alkyne (13 mM, 1 eq) in water/N,N-diisopropylethylamine (7:1, v/v) in an amber flask. The reaction mixture was stirred at 65° C. until the starting material was completely consumed or no further reaction progress was observed. The solvent was removed through high vacuum evaporation with a speedvac concentrator. The residue was dissolved in water (1 mL) and washed with CH₂Cl₂(3×). The aqueous phase was evaporated under reduced pressure using a rotary evaporator, the residue was redissolved in water, subjected to preparative reversed phase hplc and desalted, giving the triazole-containing product.
General Procedure T:
General Procedure S was followed, using Cu(PPh₃)₃Br (0.05 eq), the corresponding azide-substituted cGMP analogue (23 mM, 1 eq) and the corresponding bis-alkyne (12 mM, 2 eq) in water/N,N-diisopropylethylamine (8:1, v/v). Conditions were chosen to obtain both the monomeric and the dimeric triazole-containing product.
General Procedure U:
General Procedure S was followed, using [Cu(PPh₃)₃Br] (0.05 eq), the corresponding azide-substituted cGMP analogue (33 mM, 1 eq) and the corresponding bis-alkyne (16 mM, 0.5 eq) in water/N,N-diisopropylethylamine (10:1, v/v) to obtain the dimeric triazole-containing product.
Experimental Procedure for the Transformation of Azido-Substituted Guanosine-3′,5′-Cyclic Monophosphate Analogues into the Corresponding Amines
General Procedure V:
In a typical experiment a solution of the azido-substituted cGMP analogue (2.5 mM in water, 1 eq) in an amber flask was adjusted to pH 10 by addition of triethylamine and cooled to 10° C. DL-Dithiothreitol (5 eq) was added and the reaction mixture was stirred until the azide starting material was completely consumed or no further reaction progress was observed (usually <20 min). The mixture was evaporated to dryness under reduced pressure using a rotary evaporator. The residue was dissolved in water (1 mL), subjected to preparative reversed phase hplc and desalted, giving the amine-substituted cGMP analogue.
Experimental Procedures for the Transformation of 8-Azido-Substituted Guanosine-3′,5′-Cyclic Monophosphate Analogues into the Corresponding Iminophosphoranyl Analogue
General Procedure W:
In a typical experiment PPh₃(1.75 eq) and water (100 μL) were added to a solution of 8-azido-substituted cGMP analogue (100 mM, 1 eq) in DMF in an amber flask. The reaction mixture was stirred until the azide starting material was completely consumed or no further reaction progress was observed. The solvent was removed through high vacuum evaporation with a speedvac concentrator. The residue was suspended in water (1 mL) and washed with toluene (5×). The aqueous phase was evaporated under reduced pressure using a rotary evaporator, the residue was redissolved in water/MeOH (4:1), subjected to preparative reversed phase hplc and desalted, giving the iminophosphoranyl analogue.
Experimental Procedure for the Suzuki Cross-Coupling of Br-Substituted Guanosine-3′,5′-Cyclic Monophosphate Analogues with Organoboronic Acids
General Procedure X:
In a typical experiment aqueous K₂CO₃(2 M, 3 eq) and Pd(dppf)C12 (0.05 eq) were added successively to a solution of the Br-substituted cGMP analogue (52 mM, 1 eq) and the boronic acid (72 mM, 1.4 eq) in EtOH/H₂O (1:1, v/v). The reaction mixture was immediately degased applying three cycles of freeze-pump-thaw technique and stirred at 90° C. under argon until the bromide starting material was completely consumed or no further reaction progress was observed. The solvent was removed through high vacuum evaporation with a speedvac concentrator. The residue was suspended in water and washed with CHCl₃(3×). Methanol was added until dissolution of the precipitate (up to H₂O/MeOH=1:1). If an organic phase, containing residual CHCl₃, emerged from this composition, it was separated. The aqueous phase was then filtered through a Macherey-Nagel Chromafix C 18 (S) 270 mg cartridge (preconditioned with 10 mL of MeOH, 50% MeOH and 30% MeOH respectively) and rinsed with 30% MeOH (6 mL). The solvent was removed under reduced pressure using a rotary evaporator. The residue was dissolved in water (1 mL), subjected to preparative reversed phase hplc and desalted, giving the cross-coupling product. * All solvents used, were degassed through sonification under reduced pressure prior to the experiment.
General Procedure X2 (Preparation of Bis Boronic Acid Reagent 4-B(OH)₂PhS-PEG₅-(CH₂)₂-4-SPhB(OH)₂:
In a typical experiment N,N-diisopropylethylamine (2 eq) was added to a solution of 4-mercaptophenylboronic acid (0.2 M, 1 eq) and Br-(EO)₅—(CH₂)₂—Br (0.5 eq) in DMF. The reaction mixture was stirred until the boronic acid starting material was completely consumed or no further reaction progress was observed. The solvent was removed through high vacuum evaporation with a speedvac concentrator. The residue was dissolved in methanol (1 mL) and subjected to preparative reversed phase hplc (62% MeOH) giving 4-B(OH)₂PhS-PEG₅-(CH₂)₂-4-SPhB(OH)₂(34% yield).
Experimental Procedure for the Preparation of 1, N²-Functionalized Guanosine-3′,5′-Cyclic Monophosphate Analogues
General Procedure Y:
In a typical experiment DBU (7 eq) and the corresponding 2-bromo-aceto-reactant (3.5 eq) were added successively to a solution of the corresponding cGMP analogue (50 mM, 1 eq) in DMSO. The reaction mixture was stirred under exclusion of light until the cGMP analogue starting material was completely consumed or no further reaction progress was observed. Water (100 μL) was added, stirring was continued for 10 min and the solvent was removed through high vacuum evaporation with a speedvac concentrator. The residue was dissolved in methanol (0.5 mL) and the pH adjusted to 6-7 with HCl (1 M). In case a precipitate was formed thereby, methanol was added to redissolve it. Otherwise, water was slowly added until all components just remained soluble (max. H₂O/MeOH=5:1). The solution was subjected to preparative reversed phase hplc and desalted, giving the 1, N²-etheno-functionalized cGMP analogue.
General Procedure Y2:
In a typical experiment DBU (2 eq) and the corresponding alkyl bromoacetate-reactant (1.1 eq) were added successively to a solution of the corresponding cGMP analogue (100 mM, 1 eq) in DMSO. The reaction mixture was stirred until the cGMP analogue starting material was completely consumed or no further reaction progress was observed. Water (100 μL) was added, stirring was continued for 10 min and the solvent was removed through high vacuum evaporation with a speedvac concentrator. The residue was dissolved in H₂O (0.5 mL) and the pH adjusted to 6-7 with HCl (1 M). The solution was subjected to preparative reversed phase hplc and desalted, giving the 1, N²-acyl-functionalized cGMP analogue.
General Procedure Y3:
In a typical experiment N,N-diisopropylethylamine (2 eq) and PyBOP (1.1 eq) were added successively to a solution of the corresponding 1-carboxyalkyl-substituted cGMP analogue (10 mM in DMSO, 1 eq). The reaction mixture was stirred until the cGMP analogue starting material was completely consumed or no further reaction progress was observed. Water (100 μL) was added, stirring was continued for 10 min and the solvent was removed through high vacuum evaporation with a speedvac concentrator. The residue was dissolved in water (1 mL), the pH adjusted to 5-6 with NaOH (2 M) and the solution washed with ethyl acetate (5×). The aqueous phase was evaporated under reduced pressure using a rotary evaporator, redissolved in water, subjected to preparative reversed phase hplc and desalted, giving the 1, N²-acyl-functionalized cGMP analogue.
Experimental Procedures for the Preparation of 1-Substituted Guanosine-3′,5′-Cyclic Monophosphate Analogues
General Procedure Z:
In a typical experiment DBU (4 eq) and the corresponding bromide- (or iodide) reactant (4 eq) were added successively to a solution of the corresponding cGMP analogue (50-300 mM, 1 eq) in DMSO. The reaction mixture was stirred until the cGMP analogue starting material was completely consumed or no further reaction progress was observed. The solvent was removed through high vacuum evaporation with a speedvac concentrator. The residue was dissolved in H₂O (0.5 mL) and, in case the resulting solution was not neutral, the pH was adjusted to 7 with HCl (1 M). The solution was washed with ethyl acetate (4×). The aqueous phase was evaporated under reduced pressure using a rotary evaporator, the residue was redissolved in water, subjected to preparative reversed phase hplc and desalted, giving the 1-substituted cGMP analogue.
General Procedure Z2:
In a typical experiment DBU (2 eq) and the corresponding dibromide-reactant (0.5 eq) were added successively to a solution of the corresponding cGMP analogue (15 mM, 1 eq) in DMSO. The reaction mixture was stirred at 90° C. until the cGMP analogue starting material was completely consumed or no further reaction progress was observed. The solvent was removed through high vacuum evaporation with a speedvac concentrator. The residue was dissolved in H₂O (0.5 mL), the pH adjusted to 5-7 with HCl (1 M) and the solution was washed with ethyl acetate (4×). The aqueous phase was evaporated under reduced pressure using a rotary evaporator, the residue was redissolved in water, subjected to preparative reversed phase hplc and desalted, giving the 1-substituted dimeric cGMP analogue.
Experimental Procedures for the Preparation of 2′-OR Substituted Guanosine-3′,5′-Cyclic Monophosphate Analogues
General Procedure ZZ:
In a typical experiment 2-azidoacetic anhydride (32 eq, prepared according to Freudenberg¹³) and 4-dimethylaminopyridine (1 eq) were added successively to the corresponding cGMP analogue (1 eq). The reaction mixture was stirred under exclusion of light until the cGMP analogue starting material was completely consumed or no further reaction progress was observed. The mixture was separated between MTBE and H₂O, the organic phase was decanted off and the aqueous phase washed with MTBE (2×). A precipitate was dissolved by adding methanol to the aqueous layer. The solution was subjected to preparative reversed phase hplc and desalted, giving the 2′-O-(2-azidoacetyl)-substituted cGMP analogue.
Synthesis of previously established precursors is described in the following:

8-Carboxymethylthioguanosine-3′, 5′-cyclic Monophosphate (8-CMT-cGMP)

Using general procedure C, 8-Br-cGMP was reacted with mercaptoacetic acid to give the title compound.
Yield (Purity): 66% (>99%).
HPLC: (5% MeCN, 100 mM TEAF buffer, pH 6.8).
UV-VIS: λ_max=275 nm (pH 7), ε=13700 (est.).
ESI-MS (+): m/z calculated for C₁₂H₁₅N₅O₉PS ([M+H]⁺): 436.03, found: 436.
ESI-MS (−): m/z calculated for C₁₂H₁₃N₅O₉PS ([M−H]⁻): 434.02, found: 434.

8-Methylthioguanosine-3′, 5′-cyclic Monophosphate (8-MeS-cGMP)

Using general procedure A, 8-Br-cGMP (1 eq) was reacted with sodium methanethiolate (4 eq) to give the title compound.
Yield (Purity): 50% (>99%).
HPLC: (9 MeCN, 20 mM TEAF buffer, pH 6.8).
UV-VIS: λ_max=274 nm (pH 7), ε=14000 (est.).
ESI-MS (+): m/z calculated for C₁₁H₁₅N₅O₇PS ([M+H]⁺): 392.04, found: 392.
ESI-MS (−): m/z calculated for C₁₁H₁₃N₅O₇PS ([M−H]⁻): 390.03, found: 390.

8-Carboxyethylthioguanosine-3′, 5′-cyclic Monophosphate (8-CET-cGMP)

The title compound was synthesized from 8-MPT-cGMP using general procedure F.
Yield (Purity): 56% (>99%).
HPLC: (5 MeCN, 10 mM TEAF buffer, pH 6.8).
UV-VIS: λ_max=275 nm (pH 7), ε=13700 (est.).
ESI-MS (+): m/z calculated for C₁₃H₁₇N₅O₉PS ([M+H]⁺): 450.05, found: 450.
ESI-MS (−): m/z calculated for C₁₃H₁₅N₅O₉PS ([M−H]⁻): 448.33, found: 448.

8-(4-Thiophenylthio)guanosine-3′, 5′-cyclic Monophosphate (8-pTPT-cGMP)

Using general procedure B, 8-Br-cGMP was reacted with 1,4-benzenedithiol to give the title compound.
Yield (Purity): 41% (>99%).
HPLC: (32% MeOH, 10 mM TEAF buffer, pH 6.8).
UV-VIS: λ_max=286 nm (pH 7), ε=21500 (est.).
ESI-MS (+): m/z calculated for C₁₆H₁₇N₅O₇PS₂([M+H]+): 486.03, found: 486.
ESI-MS (−): m/z calculated for C₁₆H₁₅N₅O₇PS₂([M−H]−): 484.02, found: 484.
The invention is further illustrated by the figures and examples of Table 15 describing preferred embodiments of the present invention which are, however, not intended to limit the invention in any way. Structural examples of novel compounds are depicted in the free acid form. After HPLC workup, compounds are obtained as salts of the applied buffer, but can be transformed to other salt forms or to the free acid by cation exchange according to standard procedures for nucleotides.

TABLE 15

Examples of novel polymer linked multimeric cGMP compounds.

#	Compound/Structure

1
	Guanosine-3′,5′-cyclic monophosphate-[8-thio-(pentaethoxy)-ethylthio-8]-
	guanosine-3′,5′-cyclic monophosphate (cGMP-8-T-(EO)₅-ET-8-cGMP)
	Using general procedure E, 8-T-cGMP was reacted with Br-PEG₅-CH₂CH₂Br to give the title
	compound.
	Yield (Purity): 51% (>99%).
	HPLC: (30% MeOH, 10 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 274 nm (pH 7), ϵ = 24660 (est.).
	ESI-MS (+): m/z calculated for C₃₂H₄₆N₁₀O₁₉P₂S₂Na ([M + Na]⁺): 1023.18, found: 1023.
	ESI-MS (−): m/z calculated for C₃₂H₄₅N₁₀O₁₉P₂S₂([M − H]⁻): 999.18, found: 999.

2

	Guanosine-3′,5′-cyclic monophosphate-[8-thiomethylamidomethyl-(pentaethoxy)-
	propylamidomethylthio-8]-guanosine-3′,5′-cyclic monophosphate (cGMP-8-
	TMAmdM-(EO)₅-PrAmdMT-8-cGMP)
	Using general procedure I, 8-CMT-cGMP was reacted with NH₂CH₂-PEG₅-(CH₂)₃NH₂to
	give the title compound.
	Yield (Purity): 42% (>99%).
	HPLC: (27% MeOH, 10 mM TEAF buffer pH 6.8).
	UV-VIS: λ_max= 275 nm (pH 7), ϵ = 24660 (est.).
	ESI-MS (+): m/z calculated for C₃₈H₅₄N₁₂O₂₁P₂S₂Na₃([M − 2H + 3Na]⁺): 1209.21, found: 1209.
	ESI-MS (−): m/z calculated for C₃₈H₅₅N₁₂O₂₁P₂S₂([M − H]⁻): 1141.25, found: 1141.

3

	Guanosine-3′,5′-cyclic monophosphate-[8-thiomethylamido-(octaethoxy)-
	ethylamidomethylthio-8]-guanosine-3′,5′-cyclic monophosphate (cGMP-8-TMAmd-
	(EO)₈-EAmdMT-8-cGMP)
	Using general procedure I, 8-CMT-cGMP was reacted with NH₂-(EO)₈-(CH₂)₂NH₂to give the
	title compound.
	Yield (Purity): 38% (>98%).
	HPLC: (27% MeOH, 10 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 275 nm (pH 7), ϵ = 24660 (est.).
	ESI-MS (+): m/z calculated for C₄₂H₆₄N₁₂O₂₄P₂S₂Na ([M + Na]⁺): 1269.30, found: 1269.
	ESI-MS (−): m/z calculated for C₄₂H₆₃N₁₂O₂₄P₂S₂([M − H]⁻): 1245.30, found: 1245.

4

	Guanosine-3′,5′-cyclic monophosphate-[8-(4-thiophenylthio)-(pentaethoxy)-ethyl-
	(4-thiophenylthio)-8]-guanosine-3′,5′-cyclic monophosphate (cGMP-8-pTPT-(EO)₅-
	EpTPT-8-cGMP)
	Using general procedure E, 8-pTPT-cGMP was reacted with Br-PEG₅-CH₂CH₂Br to give the
	title compound.
	Yield (Purity): 31% (>99%).
	HPLC: (52% MeOH, 10 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 275 nm (pH 7), ϵ = 38700 (est.).
	ES-MS (+): m/z calculated for C₄₄H₅₄N₁₀O₁₉P₂S₄Na ([M + H]⁺): 1239.18, found 1239.
	ES-MS (−): m/z calculated for C₄₄H₅₃N₁₀O₁₉P₂S₄([M − H]⁻): 1215.18, found 1215.

5

	β-Phenyl-1,N²-ethenoguanosine-3′,5′-cyclic monophosphate-[8-thiomethylamido-
	(octaethoxy)-ethylamidomethylthio-8]-β-phenyl-1,N²-ethenoguanosine-3′,5′-cyclic
	monophosphate (PET-cGMP-8-TMAmd-(EO)₈-EAmdMT-8-cGMP-PET)
	Using general procedure I, 8-CMT-PET-cGMP was reacted with NH₂-PEG₈-(CH₂)₂NH₂to
	give the title compound.
	Yield (Purity): 24% (>99%).
	HPLC: (49% MeOH, 10 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 272 nm (pH 7), ϵ = 72000 (est.).
	ESI-MS (+): m/z calculated for C₅₈H₇₅N₁₂O₂₄P₂S₂([M + 2H]²⁺): 724.70, found: 724.
	ESI-MS (−): m/z calculated for C₅₈H₇₁N₁₂O₂₄P₂S₂([M − 2H]²⁻): 722.68, found: 722.

6

	8-Bromoquanosine-3′,5′-cyclic monophosphate-[1,N²-etheno-β-phenyl-4-yl-(1-
	[1,2,3]-triazole-4-yl)-methoxy-(hexaethoxy)-methyl-(4-(4-[1,2,3]-triazole-1-yl)-
	β-phenyl-1,N²-etheno)]-8-bromoguanosine-3′,5′-cyclic monophosphate (8-Br-
	cGMP-ETP-p(1-[1,2,3]-Tz-4)-MeO-(EO)₆-Me-p(4-[1,2,3]-Tz-1)-PET-cGMP-8-Br)
	Using general procedure U, 4-N₃-PET-8-Br-cGMP was reacted with Bis-Propargyl-PEG₇to
	give the title compound.
	Yield (Purity): 34% (>99%).
	HPLC: (20% MeCN, 50 mM NaH₂PO₄buffer, pH 6.8).
	UV-VIS: λ_max= 270 nm (pH 7), ϵ = 72000 (est.).
	ESI-MS (+): m/z calculated for C₅₄H₅₉Br₂N₁₆O₂₁P₂([M + H]⁺): 1487.19, found: 1487.
	ESI-MS (−): m/z calculated for C₅₄H₅₇Br₂N₁₆O₂₁P₂([M − H]⁻): 1485.17, found: 1485.

7

	Guanosine-3′,5′-cyclic monophosphate-[8-thiomethylamido-(octaethoxy)-
	ethylamidomethylthio-8]-β-phenyl-1,N₂-ethenoguanosine-3′,5′-cyclic
	monophosphate (cGMP-8-TMAmd-(EO)₈-EAmdMT-8-cGMP-PET)
	Using general procedure J, 8-CMT-PET-cGMP (C 169, 1 eq) was reacted with 8-AE-(EO)₈-
	AmdMT-cGMP (A 240, 1 eq) to give the title compound.
	Yield (Purity): 47% (>99%).
	HPLC: (19% MeCN, 50 mM NaH₂PO₄buffer, pH 6.8).
	UV-VIS: λ_max= 274 nm (pH 7), ϵ = 48330 (est.).
	ESI-MS (+): m/z calculated for C₅₀H₆₉N₁₂O₂₄P₂S₂([M + H]⁺): 1347.35, found: 1347.
	ESI-MS (−): m/z calculated for C₅₀H₆₇N₁₂O₂₄P₂S₂([M − H]⁻): 1345.33, found: 1345.

8

	Guanosine-3′,5′-cyclic monophosphate-[8-thiomethylamido-(nonadecaethoxy)-
	ethylamidomethylthio-8]-guanosine-3′,5′-cyclic monophosphate (cGMP-8-TMAmd-
	(EO)₁₉-EAmdMT-8-cGMP)
	Using general procedure L, 8-CMT-cGMP was reacted with NH₂-PEG₁₉-(CH₂)₂NH₂to give
	the title compound.
	Yield (Purity): 47% (>97%).
	HPLC (17% MeCN, 25 mM NaH₂PO₄buffer, pH 6.8).
	UV-VIS: λ_max= 275 nm (pH 7), ϵ = 24660 (est.).
	ESI-MS (+): m/z calculated for C₆₄H₁₀₉N₁₂O₃₅P₂S₂([M + H]⁺): 1731.60, found: 1732.
	ESI-MS (−): m/z calculated for C₆₄H₁₀₇N₁₂O₃₅P₂S₂([M − H]⁻): 1729.56, found: 1730.

9

	Guanosine-3′,5′-cyclic monophosphate-[8-(1-[1,2,3]-triazole-4-yl)-methoxy-
	(hexaethoxy)-methyl-(4-[1,2,3]-triazole-1-yl)-8]-guanosine-3′,5′-cyclic
	monophosphate (cGMP-8-(1-[1,2,3]-Tz-4-)-MeO-(EO)₆-Me-(4-[1,2,3]-Tz-1)-8-cGMP)
	Using general procedure U, 8-N₃-cGMP was reacted with Bis-Propargly-PEG₇to give the
	title compound.
	Yield (Purity): 4% (>99%).
	HPLC: (13% MeCN, 50 mM NaH₂PO₄buffer, pH 6.8).
	UV-VIS: λ_max= 277 nm (pH 7), ϵ = 38700 (est.).
	ESI-MS (+): m/z calculated for C₃₈H₅₃N₁₆O₂₁P₂([M + H]⁺): 1131.30, found: 1131.
	ESI-MS (−): m/z calculated for C₃₈H₅₁N₁₆O₂₁P₂([M − H]⁻): 1129.29, found: 1129.

10

	Guanosine-3′,5′-cyclic monophosphate-[8-thiomethylamido-(PEG pd 2000)-
	amidomethylthio-8]-guanosine-3′,5′-cyclic monophosphate (cGMP-8-TMAmd-(PEG
	pd 2000)-AmdMT-8-cGMP)
	Using general procedure L, 8-CMT-cGMP was reacted with NH₂-PEG_n-(CH₂)₂NH₂
	(2000 Da, polydispers) to give the title compound.
	Yield (Purity): 47% (>95%).
	HPLC: (Gradient, 21% then 24% MeCN, 50 mM NaH₂PO₄buffer, pH 6.8).
	UV-VIS: λ_max= 275 nm (pH 7), ϵ = 24660 (est.).
	ESI-MS: (−): m/z calculated for C₁₁₄H₂₀₆N₁₂O₆₀P₂S₂(n = 44, [M − 2H]²⁻): 1414.62, found: 1414.

11

	β-Phenyl-1,N²-ethenoguanosine-3′,5′-cyclic monophosphate-[8-thiomethylamido-
	(nonadecaethoxy)-ethylamidomethylthio-8]-guanosine-3′,5′-cyclic monophosphate
	(PET-cGMP-8-TMAmd-(EO)₁₉-EAmdMT-8-cGMP)
	Using the procedure J, 8-CMT-cGMP (1 eq) and 8-CMT-PET-cGMP (1 eq) were
	reacted with NH₂-PEG₁₉-(CH₂)₂NH₂in the presence of N,N-diisopropylethylamine (4 eq) and
	PyBOP (2.05 eq) to give the title compound. Conditions were chosen to additionally obtain
	the symmetrically substituted dimeric analogues.
	Yield (Purity): 35% (>99%).
	HPLC: (Gradient, 21% then 23 the 24% MeCN, 50 mM NaH₂PO₄buffer, pH 6.8).
	UV-VIS: λ_max= 274 nm (pH 7), ϵ = 49700 (est.).
	ESI-MS (+): m/z calculated for C₇₂H₁₁₃N₁₂O₃₅P₂S₂([M + H]⁺): 1831.63, found 1832.
	ESI-MS (−): m/z calculated for C₇₂H₁₁₁N₁₂O₃₅P₂S₂([M − H]⁻): 1829.62, found: 1830.

12

	β-Phenyl-1,N₂-ethenoguanosine-3′,5′-cyclic monophosphate-[8-thiomethylamido-
	(nonadecaethoxy)-ethylamidomethylthio-8]-β-phenyl-1,N²-ethenoguanosine-3′,5′-
	cyclic monophosphate (PET-cGMP-8-TMAmd-(EO)₁₉-EAmdMT-8-cGMP-PET)
	Using general procedure J, 8-CMT-cGMP (1 eq) and 8-CMT-PET-cGMP (1 eq) were
	reacted with NH₂-PEG₁₉-(CH₂)₂NH₂in the presence of N,N-diisopropylethylamine (4 eq) and PyBOP (2.05 eq) to give
	the title compound. Conditions were chosen to additionally obtain the symmetrically substituted dimeric analogues.
	Yield (Purity): 22% (>99%).
	HPLC: (Gradient, 21% then 23 then 24% MeCN, 50 mM NaH₂PO₄buffer, pH 6.8).
	UV-VIS: λ_max= 272 nm (pH 7), ϵ = 72000 (est.).
	ESI-MS (+): m/z calculated for C₈₀H₁₁₇N₁₂O₃₅P₂S₂([M + H]⁺): 1931.67, found: 1932.
	ESI-MS (−): m/z calculated for C₈₀H₁₁₅N₁₂O₃₅P₂S₂([M − H]⁻): 1929.65, found: 1930.

13

	β-Phenyl-1,N²-ethenoguanosine-3′,5′-cyclic monophosphate-[8-thiomethylamido-
	(PEG pd 2000)-amidomethylthio-8]-β-phenyl-1,N²-ethenoguanosine-3′,5′-cyclic
	monophosphate (PET-cGMP-8-TMAmd-(PEG pd 2000)-AmdMT-8-cGMP-PET)
	Using general procedure L, 8-CMT-PET-cGMP was reacted with NH₂-PEG_n-(CH₂)₂NH₂
	(2000 Da, polydispers). More PyBOP (0.5 eq) was added stepwise to drive the reaction to
	completion and yield the title compound.
	Yield (Purity): 35% (>95%).
	HPLC: (28% MeCN, 25 mM NaH₂PO₄buffer, pH 6.8).
	UV-VIS: λ_max= 272 nm (pH 7), ϵ = 72000 (est.).
	ESI-MS: (−): m/z calculated for C₁₃₀H₂₁₄N₁₂O₆₀P₂S₂(n = 44, [M − 2H]²⁻): 1516.6, found: 1516.

14

	Benzene-1,3,5-tri-[(8-amidomethyl-(pentaethoxy)-
	propylamidomethylthio)guanosine-3′,5′-cyclic monophosphate] (Bn-1,3,5-tri(AmdPr-
	(OE)₅-MAmdMT-8-cGMP)
	Using general procedure M, 8-APr-(EO)₅-MAmdMT-cGMP was reacted with 1,3,5-
	benzenetricarboxylic acid to give the title compound.
	Yield (Purity): 60% (>99%).
	HPLC: (18% MeCN, 15 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 274 nm (pH 7), ϵ = 36990 (est.).
	ESI-MS (+): m/z calculated for C₈₇H₁₃₃N₂₁O₄₂P₃S₃([M + H]⁺): 2332.73, found 2333.
	ESI-MS (−): m/z calculated for C₈₇H₁₃₁N₂₁O₄₂P₃S₃([M − H]⁻): 2330.71, found: 2331.

15

	Ethylene glycol-bis(2-aminoethylether)-N,N,N′,N′-tetra-[(8-
	methylamidoethylthio)guanosine-3′,5′-cyclic monophosphate] (EG-N,N,N′,N′-tetra(8-
	MAmdET-cGMP))
	Using general procedure N, 8-AET-cGMP was reacted with Ethylene glycol-bis(2-
	aminoethylether)-N,N,N′,N′-tetraacetic acid (EGTA) to give the title compound.
	Yield (Purity): 15% (>99%).
	HPLC: (28% MeOH, 10 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 274 nm (pH 7), ϵ = 49320 (est.).
	ESI-MS (+): m/z calculated for C₆₂H₈₅N₂₆O₃₄P₄S₄([M + H]⁺): 1989.36, found: 1989.
	ESI-MS (−): m/z calculated for C₆₂H₈₃N₂₆O₃₄P₄S₄([M − H]⁻): 1987.34, found: 1987.

16

	Guanosine-3′,5′-cyclic monophosphate-[8-thio(dodecanyl)-thio-8]-guanosine-
	3′,5′-cyclic monophosphate (cGMP-8-T-(CH₂)₁₂-T-8-cGMP)
	Using general procedure E, 8-T-cGMP was reacted with 1,12-dibromdodecane to give the
	title compound.
	Yield (Purity): 73% (>99%).
	HPLC: (26% MeCN, 30 mM NaH₂PO₄buffer, pH 6.8).
	UV-VIS: λ_max= 276 nm (pH 7), ϵ = 24660 (est.).
	ESI-MS (+): m/z calculated for C₃₂H₄₇N₁₀O₁₄P₂S₂([M + H]⁺): 921.22, found: 921.
	ESI-MS (−): m/z calculated for C₃₂H₄₅N₁₀O₁₄P₂S₂([M − H]⁻): 919.20, found: 919.

17

	β-Phenyl-1,N²-ethenoguanosine-3′,5′-cyclic monophosphate-[8-thio-(dodecanyl)-
	thio-8]-β-phenyl-1,N²-ethenoguanosine-3′,5′-cyclic monophosphate (PET-cGMP-
	8-T-(CH₂)₁₂-T-8-cGMP-PET)
	Using general procedure E, PET-8-T-cGMP was reacted with 1,12-dibromdodecane to give
	the title compound.
	Yield (Purity): 24% (>99%).
	HPLC: (36% MeCN, 20 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 275 nm (pH 7), ϵ = 72000 (est.).
	ESI-MS (−): m/z calculated for C₄₈H₅₃N₁₀O₁₄P₂S₂([M − H]⁻): 1019.27, found: 1019.

18

	Guanosine-3′,5′-cyclic monophosphate-[8-thioethylamidomethyl-(1-[1,2,3]-
	triazole-4-yl)-methoxy-(hexaethoxy)-methyl-(4-[1,2,3]-triazole-1-yl)-
	methylamidoethylthio-8]-guanosine-3′,5′-cyclic monophosphate (cGMP-8-TEAmdM-
	(1-[1,2,3]-Tz-4)-MeO-(EO)₆-Me-(4-[1,2,3]-Tz-1)-MAmdET-8-cGMP)
	Using general procedure S, 8-N3-MAmdET-cGMP (1 eq) was reacted with bis-propargyl-
	PEG₇(2 eq) to give the title compound. Conditions were chosen to additionally obtain the
	pegylated monomeric analogue.
	Yield (Purity): 23% (>99%).
	HPLC: (14% MeCN, 30 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 274 nm (pH 7), ϵ = 24660 (est.).
	ESI-MS (+): m/z calculated for C₄₆H₆₇N₁₈O₂₃P₂S₂([M + H]⁺): 1365.35, found: 1365.
	ESI-MS (−): m/z calculated for C₄₆H₆₅N₁₈O₂₃P₂S₂([M − H]⁻): 1363.34, found: 1363.

19

	Guanosine-3′,5′-cyclic monophosphate-[8-thioethylthio-8]-guanosine-3′,5′-cyclic
	monophosphate (cGMP-8-TET-cGMP)
	Using general method Q, the title compound was obtained beside monomeric azide
	analogue starting from 8-T-cGMP.
	Yield (Purity): 19% (>99%).
	HPLC: (10% MeOH, 15 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 273 nm (pH 7), ϵ = 24660 (est.).
	ESI-MS (+): m/z calculated for C₂₂H₂₇N₁₀O₁₄P₂S₂([M + H]⁺): 78.06, found: 781.
	ESI-MS (−): m/z calculated for C₂₂H₂₅N₁₀O₁₄P₂S₂([M − H]⁻): 779.05, found: 779.

20

	Guanosine-3′,5′-cyclic monophosphate-[8-thioethyl-(1-[1,2,3]-triazole-4-yl)-
	methoxy-(hexaethoxy)-methyl-(4-[1,2,3]-triazole-1-yl)-ethylthio-8]-guanosine-
	3′,5′-cyclic monophosphate (cGMP-8-TE-(1-[1,2,3]-Tz-4)-MeO-(EO)₆-Me-(4-[1,2,3]-Tz-
	1)-ET-8-cGMP)
	Using general procedure U, 8-N₃-ET-cGMP was reacted with bis-propargyl-PEG₇to give the
	title compound.
	Yield (Purity): 61% (>99%).
	HPLC: (14% MeCN, 20 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 276 nm (pH 7), ϵ = 24660 (est.).
	ESI-MS: (+): m/z calculated for C₄₂H₆₁N₁₆O₂₁P₂S₂([M + H]⁺): 1251.31, found: 1251.
	ESI-MS: (−): m/z calculated for C₄₂H₅₉N₁₆O₂₁P₂S₂([M − H]⁻): 1249.30, found: 1249.

21

	Guanosine-3′,5′-cyclic monophosphate-[8-thio-(dodecanyl)-(4-thiophenyl-4″-
	thiophenylthio)-(dodecanyl)-thio-8]-guanosine-3′,5′-cyclic monophosphate
	(cGMP-8-T-(CH₂)₁₂-pTPpTPT-(CH₂)₁₂-T-8-cGMP)
	A solution of 8-T-cGMP (200 mM, 1 eq) and N,N-diisopropylethylamine (2 eq) in DMSO was
	added portionwise over 30 min to a solution of 1,12-dibromdodecane (1.5M, 15 eq) in
	DMSO at 40′ C°. The reaction mixture was stirred until no further reaction progress was
	observed (<10% remaining starting material). The solvent was removed through high
	vacuum evaporation with a speedvac concentrator. The residue was dissolved in
	MeCN/water (8:1, v/v), washed with petroleum ether (3×) and the aqueous phase
	evaporated to dryness using a rotary evaporator. The crude product was dissolved in DMF
	(115 mM). 4,4′-Thiobisbenzenthiol (0.5 eq) and N,N-diisopropylethylamine (2.2 eq) were
	added successively. The reactin mixture was stirred until the starting material was
	completely consumed. The solvent was removed through high vacuum evaporation with a
	speedvac concentrator. The residue was dissolved in water (1 mL), washed with ethyl
	acetate (3 × 1 mL), subjected to preparative reversed phase hplc and desalted.
	Yield (Purity): 26% (>99%).
	HPLC: (57% MeCN, 30 mM NaH₂PO₄buffer, pH 6.8).
	UV-VIS: λ_max= 275 nm (pH 7), ϵ = 24660 (est.).
	ESI-MS (+): m/z calculated for C₅₆H₇₉N₁₀O₁₄P₂S₅([M + H]⁺): 1337.39, found: 1337.
	ESI-MS (−): m/z calculated for C₅₆H₇₇N₁₀O₁₄P₂S₅([M − H]⁻): 1335.37, found: 1335.

22

	Guanosine-3′,5′-cyclic monophosphate-[8-thioethylamidomethyl-(1-[1,2,3]-
	triazole-4-yl)-methoxy-(hexaethoxy)-methyl-(4-(4-[1,2,3]-triazole-1-yl)-β-
	phenyl-1,N²-etheno)]-8-bromoguanosine-3′,5′-cyclic monophosphate (cGMP-8-
	TEAmdM-(1-[1,2,3]-Tz-4)-MeO-(EO)₆-Me-(4-[1,2,3]-Tz-1)-PET-8-Br-cGMP)
	Using general procedure S, 4-N₃-PET-8-Br-cGMP (1 eq) was reacted with 8-(4-(PargO-
	(EO)₆-Me)-[1,2,3]-Tz-1)-MAmdET-cGMP (1 eq) to give the title compound.
	Yield (Purity): 26% (>99%).
	HPLC: (57% MeCN, 30 mM NaH₂PO₄buffer, pH 6.8).
	UV-VIS: λ_max= 275 nm (pH 7), ϵ = 24660 (est.).
	ESI-MS (+): m/z calculated for C₅₆H₇₉N₁₀O₁₄P₂S₅([M + H]⁺): 1337.39, found: 1337.
	ESI-MS (−): m/z calculated for C₅₆H₇₇N₁₀O₁₄P₂S₅([M − H]⁻): 1335.37, found: 1335.

23

	β-Phenyl-1,N²-ethenoguanosine-3′,5′-cyclic monophosphate-[8-thioethyl-(1-[1,2,
	3]-triazole-4-yl)-methoxy-(hexaethoxy)-methyl-(4-[1,2,3]-triazole-1-yl)-
	ethylthio-8]-β-phenyl-1,N²-ethenoguanosine-3′,5′-cyclic monophosphate (PET-
	cGMP-8-TE-(1-[1,2,3]-Tz-4)-MeO-(EO)₆-Me-(4-[1,2,3]-Tz-1)-ET-8-cGMP-PET)
	Using general procedure U, PET-8-N₃-ET-cGMP (A 257) was reacted with bis-propargyl-
	PEG₇to give the title compound.
	Yield (Purity): 23% (>99%).
	HPLC: (56% MeOH, 30 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 273 nm (pH 7), ϵ = 72000 (est.).
	ESI-MS (+): m/z calculated for C₅₈H₆₉N₁₆O₂₁P₂S₂([M + H]⁺): 1451.37, found: 1451.
	ESI-MS (−): m/z calculated for C₅₈H₆₇N₁₆O₂₁P₂S₂([M − H]⁻): 1449.36, found: 1449.

24

	8-Bromoguanosine-3′,5′-cyclic monophosphate-[1-propylamidomethyl-
	(pentaethoxy)-propylamidomethylthio-8]-β-phenyl-1,N²-ethenoguanosine-3′,5′-
	cyclic monophosphate (8-Br-cGMP-1-PrAmdM-(EO)₅-PrAmdMT-8-cGMP-PET)
	Using general procedure J, 1-AM-(EO)₅-PrAmdPr-8-Br-cGMP was reacted with 8-CMT-
	PET-cGMP to give the title compound.
	Yield (Purity): 20% (>99%).
	HPLC: (48% MeOH, 50 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 272 nm (pH 7), ϵ = 50580 (est.).
	ESI-MS (+): m/z calculated for C₄₈H₆₄N₁₂O₂₁P₂SBr ([M + H]⁺): 1317.27, found: 1317.
	ESI-MS (−): m/z calculated for C₄₈H₆₂N₁₂O₂₁P₂SBr ([M − H]⁻): 1315.25, found: 1315.

25

	8-Bromoguanosine-3′,5′-cyclic monophosphate-[1-(pentaethoxy)-ethyl-1]-8-
	bromoguanosine-3′,5′-cyclic monophosphate (8-Br-cGMP-1-(EO)₅-E-1-cGMP-8-Br)
	Using general procedure Z2, 8-Br-cGMP was reacted with Br-PEG₅-(CH₂)₂-Br to give the
	title compound.
	Yield (Purity): 32% (>99%).
	HPLC: (17% MeCN, 50 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 266 nm (pH 7), ϵ = 23400 (est.).
	ESI-MS (+): m/z calculated for C₃₂H₄₅N₁₀O₁₉P₂Br₂([M + H]⁺): 1095.07, found: 1095.
	ESI-MS (−): m/z calculated for C₃₂H₄₃N₁₀O₁₉P₂Br₂([M − H]⁻): 1093.05, found: 1093.

26

	8-Bromoguanosine-3′,5′-cyclic monophosphate-[1-propylamidomethyl-
	(pentaethoxy)-propylamidopropyl-1]-8-bromoguanosine-3′,5′-cyclic
	monophosphate (8-Br-cGMP-1-PrAmdM-(EO)₅-PrAmdPr-1-cGMP-8-Br)
	Using adapted general procedure L, 8-Br-1-CPr-cGMP was reacted with NH₂CH₂-PEG₅-
	(CH₂)₃-NH₂(3 eq) to receive the title compound and the pegylated monomeric analogue.
	Yield (Purity): 7% (>99%).
	HPLC: (36% MeOH, 100 mM TEAF buffer, pH 6.8 then (after sepperation of monomer)
	44% MeOH, 100 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 267 nm (pH 7), ϵ = 29160 (est.).
	ESI-MS (+): m/z calculated for C₄₂H₆₃N₁₂O₂₁P₂Br₂([M + H]⁺): 1293.21, found: 1293.
	ESI-MS (−): m/z calculated for C₄₂H₆₁N₁₂O₂₁P₂Br₂([M − H]⁻): 1291.19, found: 1291.

27

	8-Bromoguanosine-3′,5′-cyclic monophosphate-[1-propylamidoethyl-
	(pentaethoxy)-propylamidomethylthio-8]-guanosine-3′,5′-cyclic monophosphate
	(8-Br-cGMP-1-PrAmdM-(EO)₅-PrAmdMT-8-cGMP)
	Using general procedure J, 1-AM-(EO)₅-PrAmdPr-8-Br-cGMP was reacted with 8-CMT-
	cGMP to give the title compound.
	Yield (Purity): 32% (>99%).
	HPLC: (36% MeOH, 50 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 272 nm (pH 7), ϵ = 26910 (est.).
	ESI-MS (+): m/z calculated for C₄₀H₆₀N₁₂O₂₁P₂SBr ([M + H]⁺): 1217.24, found: 1217.
	ESI-MS (−): m/z calculated for C₄₀H₅₈N₁₂O₂₁P₂SBr ([M − H]⁻): 1215.22 found: 1215.

28

	Guanosine-3′,5′-cyclic monophosphate-[8-(phenyl-4-thio)-(pentaethoxy)-ethyl-
	(4-thiophenyl)-8]-guanosine-3′,5′-cyclic monophosphate (cGMP-8-PpT-(EO)₅-
	EpTP-8-cGMP)
	Using general procedure X, 8-Br-cGMP was reacted with 4-B(OH)₂PhS-PEG₅-(CH₂)₂-4-
	SPhB(OH)₂(0.5 eq) to give the compound.
	Yield (Purity): 11% (>99%).
	HPLC: (23% MeCN, 10 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 296 nm (pH 7), ϵ = 38700 (est.).
	ESI-MS (+): m/z calculated for C₄₄H₅₅N₁₀O₁₉P₂S₂([M + H]⁺): 1153.26, found: 1153.
	ESI-MS (−): m/z calculated for C₄₄H₅₃N₁₀O₁₉P₂S₂([M − H]⁻): 1151.24 found: 1151.

29

	β-1, N²-Acetyl-guanosine-3′,5′-cyclic monophosphate-[8-thiomethylamido-
	(octaethoxy)-ethylamidomethylthio-8]-β-1,N²-acetyl-guanosine-3′,5′-cyclic
	monophosphate (β-1,N²-Ac-cGMP-8-TMAmd-(EO)₈-EAmdMT-8-cGMP-β-1,N²-Ac)
	Using general procedure C, β-1,N²-Ac-8-Br-cGMP is reacted with mercaptoacetic acid to give
	the carboxymethyl thio substituted derivative, which is transformed to the title compound by
	reaction with NH₂-(EO)₈-(CH₂)₂NH₂applying general procedure L.

30

	8-Phenylguanosine-3′,5′-cyclic monophosphate-[1,N²-etheno-β-phenyl-4-yl-(1-
	[1,2,3]-triazole-4-yl)-methoxy-(hexaethoxy)-methyl-(4-(4-[1,2,3]-triazole-1-yl)-
	β-phenyl-1,N²-etheno)]-8-phenylguanosine-3′,5′-cyclic monophosphate (8-Phe-
	cGMP-ETP-p(1-[1,2,3]-Tz-4)-MeO-(EO)₆-Me-p(4-[1,2,3]-Tz-1)-PET-cGMP-8-Phe)
	Using general procedure Y, 8-Phe-cGMP is reacted with 4-azidophenacylbromide to give
	the 4-N₃-PET substituted derivative, which is transformed to the title compound by reaction
	with with bis-propargyl-(EO)₇applying general procedure U.

Monomeric precursors of the invention and/or momomeric compounds of the invention are further illustrated by the figures and examples of Table 16 describing preferred embodiments of the present invention which are, however, not intended to limit the invention in any way. Structural examples of novel compounds are depicted in the free acid form. After HPLC workup, compounds are obtained as salts of the applied buffer, but can be transformed to other salt forms or to the free acid by cation exchange according to standard procedures for nucleotides.

TABLE 16

Examples of monomeric precursors and/or monomeric compound of the invention.

#	Compound/Structure

31

	8-Amidomethylthioguanosine-3′,5′-cyclic monophosphate (8-AmdMT-cGMP)
	Using general procedure D, 8-T-cGMP was reacted with 2-bromoacetamide to give the title
	compound.
	Yield (Purity): 44% (>99%).
	HPLC: (5% MeCN, 10 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max275 nm (pH 7), ϵ = 13700 (est.).
	ESI-MS (+): m/z calculated for C₁₂H₁₆N₆O₈PS ([M + H]⁺): 435.05, found: 435.
	ESI-MS (−): m/z calculated for C₁₂H₁₄N₆O₈PS ([M − H]⁻): 433.03, found: 433.

32

	8-(4-Boronatephenylthio)-guanosine-3′,5′-cyclic monophosphate (8-pB(OH)₂PT-
	cGMP)
	Using general procedure A, 8-Br-cGMP was reacted with 4-mercaptophenylboronic acid to
	give the title compound.
	Yield (Purity): 71% (>99%).
	HPLC: (32% MeOH, 10 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 276 nm (pH 7), ϵ = 21500 (est.).
	ESI-MS (+): m/z calculated for C₁₆H₁₇BN₅O₉PSNa ([M + H]⁺): 520.05, found: 520.
	ESI-MS (−): m/z calculated for C₁₆H₁₆BN₅O₉PS ([M − H]⁻): 496.05, found: 496.

33

	8-(4-Cyanobenzylthio)guanosine-3′,5′-cyclic monophosphate (8-pCNBT-cGMP)
	Using general procedure D, 8-T-cGMP was reacted with 4-cyanobenzyl bromide to give the
	title compound.
	Yield (Purity): 34% (>99%).
	HPLC: (32% MeOH, 10 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 276 nm (pH 7), ϵ = 13700 (est.).
	ESI-MS (+): m/z calculated for C₁₈H₁₈N₆O₇PS ([M + H]⁺): 493.07, found: 493.
	ESI-MS (−): m/z calculated for C₁₈H₁₆N₆O₇PS ([M − H]⁻): 491.05, found: 491.

34

	8-(4-(2-Cyanophenyl)-benzylthio)guanosine-3′,5′-cyclic monophosphate (8-(p(2-
	CNPhe)BT-cGMP
	Using general procedure D, 8-T-cGMP was reacted with 4-bromomethyl-2-cyanobiphenyl to
	give the title compound.
	Yield (Purity): 16% (>99%).
	HPLC: (52% MeOH, 10 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 260 nm (pH 7), ϵ = 13700 (est.).
	ESI-MS (+): m/z calculated for C₂₄H₂₂N₆O₇PS ([M + H]⁺): 569.10, found: 569.
	ESI-MS (−): m/z calculated for C₂₄H₂₀N₆O₇PS ([M − H]⁻): 567.09, found: 567.

35

	8-Cyclohexylmethylthioguanosine-3′,5′-cyclic monophosphate (8-cHeMT-cGMP)
	Using general procedure D, 8-T-cGMP (1 eq) was reacted with cyclohexylmethyl bromide
	(2 eq) at 90° C. to give the title compound.
	Yield (Purity): 38% (>99%).
	HPLC: (57% MeOH, 10 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 276 nm (pH 7), ϵ = 13700 (est.).
	ESI-MS (+): m/z calculated for C₁₇H₂₅N₅O₇PS ([M + H]⁺): 474.12, found: 474.
	ESI-MS (−): m/z calculated for C₁₇H₂₃N₅O₇PS ([M − H]⁻): 472.11, found: 472.

36

	8-(2,4-Dichlorophenylthio)guanosine-3′,5′-cyclic monophosphate (8-o,pDCIPT-
	cGMP)
	Using general procedure A, 8-Br-cGMP was reacted with 2,4-dichlorobenzenethiol to give
	the title compound.
	Yield (Purity): 33% (>99%).
	HPLC: (22% MeCN, 10 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 279 nm (pH 7), ϵ = 21500 (est.).
	ESI-MS (+): m/z calculated for C₁₆H₁₄Cl₂N₅O₇PSNa ([M + Na]⁺): 543.96, found: 544.
	ESI-MS (−): m/z calculated for C₁₆H₁₃Cl₂N₅O₇PS ([M − H]⁻): 519.97, found: 520.

37

	8-Diethylphosphonoethylthio-guanosine-3′,5′-cyclic monophosphate (8-DEPET-
	cGMP)
	Using general procedure D, 8-T-cGMP was reacted with diethyl 2-bromoethylphosphonat to
	give the title compound.
	Yield (Purity): 20% (>99%).
	HPLC: (32% MeOH, 10 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 275 nm (pH 7), ϵ = 13700 (est.).
	ESI-MS (+): m/z calculated for C₁₆H₂₆N₅O₁₀P₂S ([M + H]⁺): 542.09, found: 542.
	ESI-MS (−): m/z calculated for C₁₆H₂₄N₅O₁₀P₂S ([M − H]⁻): 540.07, found: 540.

38

	8-Ethylthioguanosine-3′,5′-cyclic monophosphate (8-ET-cGMP)
	Using general procedure A, 8-Br-cGMP was reacted with ethanethiol to give the title
	compound.
	Yield (Purity): 30% (>99%).
	HPLC: (10% MeCN, 10 mM NaH2PO4 buffer, pH 4.8).
	UV-VIS: λ_max= 275 nm (pH 7), ϵ = 14000 (est.).
	ESI-MS (+): m/z calculated for for C₁₂H₁₇N₅O₇PS ([M + H]⁺): 406.06, found: 406.
	ESI-MS (−): m/z calculated for C₁₂H₁₅N₅O₇PS ([M − H]⁻): 404.04, found: 404.

39

	8-Hexylthioguanosine-3′,5′-cyclic monophosphate (8-HT-cGMP)
	Using general procedure A, 8-Br-cGMP was reacted with 1-hexaethiol to give the title
	compound.
	Yield (Purity): 50% (>99%).
	HPLC: (21% MeCN, 20 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 276 nm (pH 7), ϵ = 14000 (est.).
	ESI-MS (+): m/z calculated for C₁₆H₂₅N₅O₇PS ([M + H]⁺): 462.12, found: 462.
	ESI-MS (−): m/z calculated for C₁₆H₂₃N₅O₇PS ([M − H]⁻): 460.11, found: 460.

40

	8-(4-Isopropylphenylthio)guanosine-3′,5′-cyclic monophosphate (8-pIPrPT-cGMP)
	Using general procedure A, 8-Br-cGMP was reacted with 4-Isopropylthiophenol to give the
	title compound.
	Yield (Purity): 75% (>98%).
	HPLC: (55% MeOH, 10 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 278 nm (pH 7), ϵ = 21500 (est.).
	ESI-MS (+): m/z calculated for C₁₉H₂₃N₅O₇PS ([M + H]⁺): 496.11, found: 496.
	ESI-MS (−): m/z calculated for C₁₉H₂₁N₅O₇PS ([M − H]⁻): 494.09, found: 494.

41

	8-(3-(2-Methyl)furanyl)thioguanosine-3′,5′-cyclic monophosphate (8-(3-(2-Me)-
	FU)T-cGMP)
	Using general procedure A, 8-Br-cGMP was reacted with 2-methyl-3-furanethiol to give the
	title compound.
	Yield (Purity): 32% (>99%).
	HPLC: (37% MeOH, 10 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 275 nm (pH 7), ϵ = 21500 (est.).
	ESI-MS (+): m/z calculated for C₁₅H₁₇N₅O₈PS ([M + H]⁺): 458.05, found: 458.
	ESI-MS (−): m/z calculated for C₁₅H₁₅N₅O₈PS ([M − H]⁻): 456.04, found: 456.

42

	8-(5-(1-Methyl)tetrazolyl)thioguanosine-3′,5′-cyclic monophosphate (8-(5-(1-Me)-
	Tet)T-cGMP
	Using general procedure A, 8-Br-cGMP was reacted with 5-mercapto-1-methyltetrazole to
	give the title compound.
	Yield (Purity): 29% (>95%).
	HPLC: (7% MeCN, 10 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 276 nm (pH 7), ϵ = 21500 (est.).
	ESI-MS (+): m/z calculated for C₁₂H₁₅N₉O₇PS ([M + H]⁺): 460.06, found: 460.
	ESI-MS (−): m/z calculated for C₁₂H₁₃N₉O₇PS ([M − H]⁻): 458.04, found: 458.

43

	8-(4-Methoxybenzylthio)guanosine-3′,5′-cyclic monophosphate (8-pMeOBT-cGMP)
	Using the general procedure A, 8-Br-cGMP (1 eq) was reacted with 4-methoxybenzyl
	mercaptan (4 eq) to give the title compound.
	Yield (Purity): 37% (>99%).
	HPLC: (21% MeCN, 20 mM TEAF buffer, pH 6.8.).
	UV-VIS: λ_max= 277 nm (pH 7), ϵ = 21500 (est.).
	ESI-MS (+): m/z calculated for C₁₈H₂₁N₅O₈PS ([M + H]⁺): 498.08, found: 498.
	ESI-MS (−): m/z calculated for C₁₈H₁₉N₅O₈PS ([M − H]⁻): 496.07, found: 496.

44

	8-(7-(4-Methyl)coumarinyl)thio-guanosine-3′,5′-cyclic monophosphate (8-(7-(4-Me)-
	Cou)T-cGMP)
	Using general procedure A, 8-Br-cGMP was reacted with 7-mercapto-4-methylcoumarin to
	give the title compound.
	Yield (Purity): 77% (>99%).
	HPLC: (35% MeOH, 10 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 281 nm (pH 7), ϵ = 21500 (est.).
	ESI-MS (+): m/z calculated for C₂₀H₁₉N₅O₉PS ([M + H]⁺): 536.06, found: 536.
	ESI-MS (−): m/z calculated for C₂₀H₁₇N₅O₉PS ([M − H]⁻): 534.05, found: 534.

45

	8-Methylacetylthioguanosine-3′,5′-cyclic monophosphate (8-MAcT-cGMP)
	Using general procedure D, 8-T-cGMP was reacted with methyl bromoacetate to give the
	title compound.
	Yield (Purity): 35% (>98%).
	HPLC: (25% MeOH, 10 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 274 nm (pH 7), ϵ = 13700 (est.).
	ESI-MS (+): m/z calculated for C₁₃H₁₇N₅O₉PS ([M + H]⁺): 450.05, found: 450.
	ESI-MS (−): m/z calculated for C₁₃H₁₅N₅O₉PS ([M − H]⁻): 448.03, found: 448.

46

	8-(5-(1-Phenyl)tetrazolyl)thioguanosine-3′,5′-cyclic monophosphate (8-(5-(1-Phe)-
	Tet)T-cGMP)
	Using general procedure A, 8-Br-cGMP was reacted with 5-mercapto-1-phenyl-1H-tetrazole
	to give the title compound.
	Yield (Purity): 44% (>99%).
	HPLC: (32% MeOH, 10 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 277 nm (pH 7), ϵ = 21500 (est.).
	ESI-MS (+): m/z calculated for C₁₇H₁₇N₉O₇PS ([M + H]⁺): 522.07, found: 522.
	ESI-MS (−): m/z calculated for C₁₇H₁₅N₉O₇PS ([M − H]⁻): 520.06, found: 520.

47

	8-(2-Phenylethyl)thioguanosine-3′,5′-cyclic monophosphate (8-PhEtT-cGMP)
	Using general procedure A, 8-Br-cGMP (1 eq) was reacted with 2-phenylethanethio (4 eq)
	to give the title compound.
	Yield (Purity): 20% (>99%).
	HPLC: (21% MeCN, TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 277 nm (pH 7), ϵ = 21500 (est.).
	ESI-MS (+): m/z calculated for C₁₈H₂₁N₅O₇PS ([M + H]⁺): 482.09, found: 482.
	ESI-MS (−): m/z calculated for C₁₈H₁₉N₅O₇PS ([M − H]⁻): 480.07, found: 480.

48

	8-(2-(4-Phenyl)imidazolyl)thioguanosine-3′,5′-cyclic monophosphate (8-(2-(4-Phe)-
	Im)T-cGMP)
	Using general procedure A, 8-Br-cGMP was reacted with 4-phenylimidazole-2-thiol to give
	the title compound.
	Yield (Purity): 26% (>99%).
	HPLC: (35% MeOH, 10 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 276 nm (pH 7), ϵ = 21500 (est.).
	ESI-MS (+): m/z calculated for C₁₉H₁₉N₇O₇PS ([M + H]⁺): 520.08, found: 520.
	ESI-MS (−): m/z calculated for C₁₉H₁₇N₇O₇PS ([M − H]⁻): 518.07, found: 518.

49

	8-(2-Thiophenyl)thioguanosine-3′,5′-cyclic monophosphate (8-(2-Tp)T-cGMP)
	Using general procedure A, 8-Br-cGMP was reacted with 2-thiophenethiol to give the title
	compound.
	Yield (Purity): 21% (>98%.)
	HPLC: (30% MeOH, 10 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 277 (pH 7), ϵ = 21500 (est.).
	ESI-MS (+): m/z calculated for C₁₄H₁₅N₅O₇PS₂([M + H]⁺): 460.02, found: 460.
	ESI-MS (−): m/z calculated for C₁₄H₁₃N₅O₇PS₂([M − H]⁻): 458.00, found: 458.

50

	8-(1,1,2-Trifluoro-1-butenthio)guanosine-3′,5′-cyclic monophosphate (8-(1,1,2-
	TF-Bu(1-en))T-cGMP)
	Using general procedure D, 8-TcGMP was reacted with 4-bromo-1,1,2-trifluoro-1-butene to
	give the title compound.
	Yield (Purity): 21% (>99%).
	HPLC: (37% MeOH, 10 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 276 nm (pH 7), ϵ = 13700 (est.).
	ESI-MS (+): m/z calculated for C₁₄H₁₆F₃N₅O₇PS ([M + H]⁺): 486.05, found: 486.
	ESI-MS (−): m/z calculated for C₁₄H₁₄F₃N₅O₇PS ([M − H]⁻): 484.03, found: 484.

51

	8-Amidopropylthioguanosine-3′,5′-cyclic monophosphate (8-AmdPrT-cGMP)
	The title compound was synthesized from 8-EButT-cGMP using general procedure G.
	Yield (Purity): 18% (>99%).
	HPLC: (32% MeOH, 10 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 0 274 nm (pH 7), ϵ = 13700 (est.).
	ES-MS (+): m/z calculated for C₁₄H₂₀N₆O₈PS ([M + H]⁺): 463.08, found: 463.
	ES-MS (−): m/z calculated for C₁₄H₁₈N₆O₈PS ([M − H]⁻): 461.06, found: 461.

52

	8-Amidoethylthioguanosine-3′,5′-cyclic monophosphate (8-AmdET-cGMP)
	The title compound was synthesized from 8-MPT-cGMP using general procedure G.
	Yield (Purity): 57% (>99%).
	HPLC: (6% MeCN, 10 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 275 nm (pH 7), ϵ = 13700 (est.).
	ESI-MS (+): m/z calculated for C₁₃H₁₈N₆O₈PS ([M + H]⁺): 449.06, found: 449.
	ESI-MS (−): m/z calculated for C₁₃H₁₆N₆O₈PS ([M − H]⁻): 447.34, found:.

53

	8-Amidobutylthioguanosine-3′,5′-cyclic monophosphate (8-AmdBuT-cGMP)
	The title compound was synthesized from 8-MVAlT-cGMP using general procedure G.
	Yield (Purity): 24% (>99%).
	HPLC: (6% MeCN, 10 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 275 nm (pH 7), ϵ = 13700 (est.).
	ESI-MS (+): m/z calculated for C₁₅H₂₂N₆O₈PS ([M + H]⁺): 477.10, found: 477.
	ESI-MS (−): m/z calculated for C₁₅H₂₀N₆O₈PS ([M − H]⁻): 475.08, found: 475.

54

	8-Acetamidoethylthioguanosine-3′,5′-cyclic monophosphate (8-AcAmdET-cGMP)
	Using general procedure H, 8-AET-cGMP was reacted with acetic acid to give the title
	compound.
	Yield (Purity): 34% (>98%).
	HPLC: (6% MeCN, 10 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 275 nm (pH 7), ϵ = 13700 (est.).
	ESI-MS (+): m/z calculated for C₁₄H₂₀N₆O₈PS ([M + H]⁺): 463.08, found: 463.
	ESI-MS (−): m/z calculated for C₁₄H₁₈N₆O₈PS ([M − H]⁻): 461.06, found:.

55

	8-(2-Benzothiazolyl)thioguanosine-3′,5′-cyclic monophosphate (8-(2-BT)T-cGMP)
	Using general procedure A, 8-Br-cGMP was reacted with 2-mercaptobenzothiazole to give
	the title compound.
	Yield (Purity): 3% (>99%).
	HPLC: (37% MeOH, 10 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 278 nm (pH 7), ϵ 21500 (est.).
	ESI-MS (+): m/z calculated for C₁₇H₁₆N₆O₇PS₂([M + H]⁺): 511.03, found: 511.
	ESI-MS (−): m/z calculated for C₁₇H₁₄N₆O₇PS₂([M + H]⁺): 509.01, found: 509.

56

	8-(2-Boronatebenzylthio)guanosine-3′,5′-cyclic monophosphate (8-(oB(OH)₂Bn)T-
	cGMP)
	Using general procedure D, 8-T-cGMP was reacted with 2-(bromomethyl)phenylboronic
	acid to give the title compound.
	Yield (Purity): 37% (>99%).
	HPLC: (32% MeOH, 10 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 278 nm (pH 7), ϵ = 13700 (est.).
	ESI-MS (+): m/z calculated for C₁₇H₁₉BN₅O₉PSNa ([M + Na]⁺): 534.06, found: 534.
	ESI-MS (−): m/z calculated for C₁₇H₁₈BN₅O₉PS ([M − H]⁻): 510.07, found: 510.

57

	8-(4-Boronatebutylthio)guanosine-3′,5′-cyclic monophosphate (8-(pB(OH)₂Bu)T-
	cGMP)
	Using general procedure D, 8-T-cGMP was reacted with 4-bromobutylboronic acid to give
	the title compound.
	Yield (Purity): 43% (>99%).
	HPLC: (30% MeOH, 10 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 275 nm (pH 7), ϵ = 13700 (est.).
	ESI-MS (+): m/z calculated for C₁₄H₂₁BN₅O₉PSNa ([M + H]⁺): 500.08, found: 500.
	ESI-MS (−): m/z calculated for C₁₄H₂₀BN₅O₉PS ([M − H]⁻): 476.08, found: 476.

58

	8-(4-Boronatebenzylthio)guanosine-3′,5′-cyclic monophosphate (8-(pB(OH)₂Bn)T-
	cGMP)
	Using general procedure D, 8-T-cGMP was reacted with 4-(bromomethyl)phenylboronic
	acid to give the title compound.
	Yield (Purity): 67% (>99%).
	HPLC: (30% MeOH, 10 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 278 nm (pH 7), ϵ = 13700 (est.).
	ESI-MS (+): m/z calculated for C₁₇H₂₀BN₅O₉PS ([M + H]⁺): 512.08, found: 512.
	ESI-MS (−): m/z calculated for C₁₇H₁₈BN₅O₉PS ([M − H]⁻): 510.07, found: 510.

59

	8-(3-Boronatebenzylthio)guanosine-3′,5′-cyclic monophosphate (8-(mB(OH)₂Bn)T-
	cGMP)
	Using general procedure D, 8-T-cGMP was reacted with 3-(bromomethyl)phenylboronic
	acid to give the title compound.
	Yield (Purity): 54% (>99%).
	HPLC: (30% MeOH, 10 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 278 nm (pH 7), ϵ = 13700 (est.).
	ESI-MS (+): m/z calculated for C₁₇H₁₉BN₅PSNa ([M + Na]⁺): 534.06, found: 534.
	ESI-MS (−): m/z calculated for C₁₇H₁₈BN₅O₉PS ([M − H]⁻): 510.07, found: 510.

60

	8-Azidomethylamidoethylthioguanosine-3′,5′-cyclic monophosphate (8-N₃-
	MAmdET-cGMP)
	Using general procedure J, 8-AET-cGMP was reacted with azidoacetic acid to give the title
	compound.
	Yield (Purity): 62% (>98%).
	HPLC: (21% MeOH, 20 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 276 nm (pH 7), ϵ = 13700 (est.).
	ESI-MS (+): m/z calculated for C₁₄H₁₉N₉O₈PS ([M + H]⁺): 504.08, found: 504.
	ESI-MS (−): m/z calculated for C₁₄H₁₇N₉O₈PS ([M − H]⁻): 502.07, found: 502.

61

	8-(3-Boronatephenyl)amidobutylthio-guanosine-3′,5′-cyclic monophosphate (8-
	(mB(OH)₂PAmdBu)T-cGMP)
	Using general procedure H, 8-CBuT-cGMP was reacted with 3-aminobenzeneboronic acid
	to give the title compound.
	Yield (Purity): 50% (>99%).
	HPLC: (35% MeOH, 10 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 276 nm (pH 7), ϵ = 13700 (est.).
	ESI-MS (−): m/z calculated for C₂₁H₂₅BN₆O₁₀PS ([M − H]⁻): 595.12, found: 595.

62

	8-Benzylamidobutylthioguanosine-3′,5′-cyclic monophosphate (8-BnAmdBuT-
	cGMP)
	Using general procedure H, 8-CBuT-cGMP was reacted with benzylamine to give the title
	compound.
	Yield (Purity): 49% (>99%).
	HPLC: (35% MeOH, 10 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 276 nm (pH 7), ϵ = 13700 (est.).
	ESI-MS (+): m/z calculated for C₂₂H₂₈N₆O₈PS ([M + H]⁺): 567.14, found: 567.
	ESI-MS (−): m/z calculated for C₂₂H₂₆N₆O₈PS ([M − H]⁻): 565.13, found: 565.

63

	8-Benzamidoethylthioguanosine-3′,5′-cyclic monophosphate (8-BAmdET-cGMP)
	Using general procedure H, 8-AET-cGMP was reacted with benzoic acid to give the title
	compound.
	Yield (Purity): 12% (>99%).
	HPLC: (25% MeOH, 10 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 277 nm (pH 7), ϵ = 13700 (est.).
	ESI-MS (+): m/z calculated for C₁₉H₂₂N₆O₈PS ([M + H]⁺): 525.10, found: 525.
	ESI-MS (−): m/z calculated for C₁₉H₂₀N₆O₈PS ([M − H]⁻): 523.08, found: 523.

64

	8-(3-Boronatephenyl)amidomethyl-thioguanosine-3′,5′-cyclic monophosphate (8-
	mB(OH)₂PAmdMT-cGMP)
	Using general procedure H, 8-CMT-cGMP was reacted with 3-aminophenylboronic acid to
	give the title compound.
	Yield (Purity): 24% (>99%).
	HPLC: (15% MeOH, 10 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 272 nm (pH 7), ϵ = 13700 (est.).
	ESI-MS (−): m/z calculated for C₁₈H₁₉BN₆O₁₀PS ([M − H]⁻): 553.07, found: 553.

65

	8-Benzylamidomethylthio-guanosine-3′,5′-cyclic monophosphate (8-BnAmdMT-
	cGMP)
	Using general procedure H, 8-CMT-cGMP was reacted with benzylamine to give the title
	compound.
	Yield (Purity): 53% (>99%).
	HPLC: (28% MeOH, 10 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 278 nm (pH 7), ϵ = 13700 (est.).
	ESI-MS (+): m/z calculated for C₁₉H₂₂N₆O₈PS ([M + H]⁺): 525.10, found: 525.
	ESI-MS (−): m/z calculated for C₁₉H₂₀N₆O₈PS ([M − H]⁻): 523.08, found: 523.

66

	8-(3-Boronatephenyl)amidoethylthio-guanosine-3′,5′-cyclic monophosphate (8-
	mB(OH)₂PAmdET-cGMP)
	Using general procedure H, 8-CET-cGMP was reacted with 3-aminophenylboronic acid to
	give the title compound.
	Yield (Purity): 11% (>99%).
	HPLC: (32% MeOH, 10 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 255 (+278) nm (pH 7), ϵ = 13700 (est.).
	ESI-MS (−): m/z calculated for C₁₉H₂₁BN₆O₁₀PS ([M − H]⁻): 567.09, found: 567.

67

	8-(3-Boronatephenyl)amidopropylthioguanosine-3′,5′-cyclic monophosphate (8-
	mB(OH)₂PAmdPrT-cGMP)
	Using general procedure H, 8-CPrT-cGMP was reacted with 3-aminophenylboronic acid to
	give the title compound.
	Yield (Purity): 65% (>99%).
	HPLC: (32% MeOH, 10 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 276 nm (pH 7), ϵ = 13700 (est.).
	ESI-MS (−): m/z calculated for C₂₀H₂₃BN₆O₁₀PS ([M − H]⁻): 581.10, found: 581.

68

	8-Carboxypropylthioguanosine-3′,5′-cyclic monophosphate (8-CPrT-cGMP)
	The title compound was synthesized from 8-EButT-cGMP using general procedure F.
	Yield (Purity): 95% (>99%).
	HPLC: (5% MeCN, 10 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 275 nm (pH 7), ϵ = 13700 (est.).
	ESI-MS (+): m/z calculated for C₁₄H₁₉N₅O₉PS ([M + H]⁺): 464.06, found: 464.
	ESI-MS (−): m/z calculated for C₁₄H₁₇N₅O₉PS ([M − H]⁻): 462.05, found: 462.

69

	8-Carboxybutylthioguanosine-3′,5′-cyclic monophosphate (8-CBuT-cGMP)
	The title compound was synthesized from 8-MVAlT-cGMP using general procedure F.
	Yield (Purity): 87% (>99%).
	HPLC: (6% MeCN, 10 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 275 nm (pH 7), ϵ = 13700 (est.).
	ESI-MS (+): m/z calculated for C₁₅H₂₁N₅O₉PS ([M + H]⁺): 478.08, found: 478.
	ESI-MS (−): m/z calculated for C₁₅H₁₉N₅O₉PS ([M − H]⁻): 476.06, found: 476.

70

	8-(2,6-Dichlorophenoxypropyl)thio-guanosine-3′,5′-cyclic monophosphase (8-(2,6-
	DClPheoPr)T-cGMP
	Using general procedure D, 8-T-cGMP was reacted with 2-(3-bromopropoxy)-1,3-
	dichlorobenzene to give the title compound.
	Yield (Purity): 21% (>99%).
	HPLC: (52% MeOH, 10 mM TEAF buffer, pH 6.8).
	US-VIS: λ_max= 275 nm (pH 7), ϵ = 13700 (est.).
	ESI-MS (+): m/z calculated for C₁₉H₂₁Cl₂N₅O₈PS ([M + H]⁺): 580.02, found: 580.
	ESI-MS (−): m/z calculated for C₁₉H₁₉Cl₂N₅O₈PS ([M − H]⁻): 578.01, found: 578.

71

	8-(4-Dimethylaminophenyl)amido-methylthioguanosine-3′,5′-cyclic
	monophosphate (8-pDMAPAmdMT-cGMP)
	Using general procedure H, 8-CMT-cGMP was reacted with N,N-dimethyl-p-
	phenylenediamine dihydrochloride applying 3.3 eq of N,N-diisopropylethylamie to give the
	title compound.
	Yield (Purity): 73% (>99%).
	HPLC: (36% MeOH, 10 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 274 nm (pH 7), ϵ = 13700 (est.).
	ESI-MS (+): m/z calculated for C₂₀H₂₅N₇O₈PS ([M + H]⁺): 554.12, found: 554.
	ESI-MS (−): m/z calculated for C₂₀H₂₃N₇O₈PS ([M − H]⁻): 552.11, found: 552.

72

	8-(4-Dimethylaminophenyl)amido-butylthioguanosine-3′,5′-cyclic monophosphate
	(8-pDMAPAmdBuT-cGMP)
	Using general procedure H, 8-CBuT-cGMP was reacted with N,N-dimethyl-p-
	phenylenediamine dihydrochloride applying 3.3 eq of N,N-diisopropylethylamie to give the
	title compound.
	Yield (Purity): 88% (>99%).
	HPLC: (36% MeOH, 10 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 270 nm (pH 7), ϵ = 13700 (est.).
	ESI-MS (+): m/z calculated for C₂₃H₃₁N₇O₈PS ([M + H]⁺): 596.17, found: 596.
	ESI-MS (−): m/z calculated for C₂₃H₂₉N₇O₈PS ([M − H]⁻): 594.15, found, 594.

73

	8-Ethylbutyrylthioguanosine-3′,5′-cyclic monophosphate (8-EButT-cGMP)
	Using general procedure D, 8-T-cGMP was reacted with ethyl 4-bromobutyrate to give to
	title compound.
	Yield (Purity): 54% (>99%).
	HPLC: (30% MeOH, 10 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 275 nm (ph 7), ϵ = 13700 (est.).
	ESI-MS: (+): m/z calculated for C₁₆H₂₃N₅O₉PS ([M + H]⁺): 492.10, found: 492.
	ESI-MS: (−): m/z calculated for C₁₆H₂₁N₅O₉PS ([M − H]⁻): 490.08, found: 490.

74

	8-Methylpropionylthioguanosine-3′,5′-cyclic monophosphate (8-MPT-cGMP)
	Using general procedure D, 8-T-cGMP was reacted with methyl 3-bromopropionate and
	equivalents were increased stepwise (methyl 3-bromopropionate up to 9 eq, N,N-
	diisopropylethylamine up to 8 eq) to improve the yield of the title compound.
	Yield (Purity): 47% (>99%).
	HPLC: (15% MeOH, 10 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 275 nm (pH 7), ϵ = 13700 (est.).
	ESI-MS (+): m/z calculated for C₁₄H₁₉N₅O₉PS ([M + H]⁺): 464.06, found: 464.
	ESI-MS (−): m/z calculated for C₁₄H₁₇N₅O₉PS ([M − H]⁻): 462.05, found: 462.

75

	8-Methylvalerianylthioguanosine-3′,5′-cyclic monophosphate (8-MValT-cGMP)
	Using general procedure D, 8-T-cGMP was reacted with methyl 5-bromovalerateto give the
	title compound.
	Yield (Purity): 75% (>99%).
	HPLC: (15% MeOH, 10 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 275 nm (pH 7), ϵ = 13700 (est.).
	ESI-MS (+): m/z calculated for C₁₆H₂₃N₅O₉PS ([M + H]⁺): 492.10, found: 492.
	ESI-MS (−): m/z calculated for C₁₆H₂₁N₅O₉PS ([M − H]⁻): 490.08, found: 490.

76

	8-Methoxyethylamidobutylthio-guanosine-3′,5′-cyclic monophosphate (8-
	MeOEAmdBuT-cGMP)
	Using general procedure H, 8-CBuT-cGMP was reacted with 2-methoxyethylamine to give
	the title compound.
	Yield (Purity): 58% (>99%).
	HPLC: (30% MeOH, 10 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 276 nm (pH 7), ϵ = 13700 (est.).
	ESI-MS (+): m/z calculated for C₁₈H₂₈N₆O₉PS ([M + H]⁺): 535.14, found 535.
	ESI-MS (−): m/z calculated for C₁₈H₂₆N₆O₉PS ([M − H]⁻): 533.12, found 533.

77

	8-Methoxyethylamidomethylthio-guanosine-3′,5′-cyclic monophosphate (8-
	MeOEAmdMT-cGMP)
	Using general procedure H, 8-CMT-cGMP was reacted with 2-methoxyethylamine to give
	the title compound.
	Yield (Purity): 65% (>99%).
	HPLC: (15% MeOH, 10 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 275 nm (pH 7), ϵ = 13700 (est.).
	ESI-MS (+): m/z calculated for C₁₅H₂₂N₆O₉PS ([M + H]⁺): 493.09, found: 493.
	ESI-MS (−): m/z calculated for C₁₅H₂₀N₆O₉PS ([M − H]⁻): 491.08, found: 491.

78

	8-Methoxyethylamidoethylthio-guanosine-3′,5′-cyclic monophosphate (8-
	MeOEAmdEt-cGMP)
	Using general procedure H, 8-CET-cGMP was reacted with 2-methoxyethylamine to give
	the title compound.
	Yield (Purity): 65% (>99%).
	HPLC: (21% MeOH, 10 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 275 nm (pH 7), ϵ = 13700 (est.).
	ESI-MS (+): m/z calculated for C₁₆H₂₄N₆O₉PS ([M + H]⁺): 507.11, found: 507.
	ESI-MS (−): m/z calculated for C₁₆H₂₂N₆O₉PS ([M − H]⁻): 505.09, found: 505.

79

	8-Phenylamidomethylthio-guanosine-3′,5′-cyclic monophosphate (8-PAmdMT-
	cGMP)
	Using general procedure H, 8-CMT-cGMP was reacted with aniline to give the title
	compound.
	Yield (Purity): 80% (>99%).
	HPLC: (32% MeOH, 10 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 273 nm (pH 7), ϵ = 13700 (est.).
	ESI-MS (+): m/z calculated for C₁₈H₂₀N₆O₈PS ([M + H]⁺): 511.08, found: 511.
	ESI-MS (−): m/z calculated for C₁₈H₁₈N₆O₈PS ([M − H]⁻): 509.06, found: 509.

80

	8-Phenylpropylthioguanosine-3′,5′-cyclic monophosphate (8-PPrT-cGMP)
	Using general procedure D, 8-T-cGMP was reacted with 1-Bromo-3-phenylpropane at 90° C.
	to give the title compound.
	Yield (Purity): 68% (>99%).
	HPLC: (49% MeOH, 10 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 276 nm (pH 7), ϵ = 13700 (est.).
	ESI-MS (+): m/z calculated for C₁₉H₂₃N₅O₇PS ([M + H]⁺): 496.11, found: 496.
	ESI-MS (−): m/z calculated for C₁₉H₂₁N₅O₇PS ([M − H]⁻): 494.09, found: 494.

81

	8-(3-Butynylthio)guanosine-3′,5′-cyclic monophosphate (8-(Bu(3-yne)T-cGMP)
	Using general procedure D, 8-T-cGMP was reacted with 4-bromo-1-butyne to give the title
	compound.
	Yield (Purity): 51% (>99%).
	HPLC: (30% MeOH, 10 mM TEAF buffer, pH 6.8).
	US-VIS: λ_max= 275 nm (pH 7), ϵ = 13700 (est.).
	ESI-MS (+): m/z calculated for C₁₄H₁₇N₅O₇PS ([M + H]⁺): 430.06, found: 430.
	ESI-MS (+): m/z calculated for C₁₄H₁₅N₅O₇PS ([M − H]⁻): 428.34, found: 428.

82

	8-(4-Acetamidophenylthio)guanosine-3′,5′-cyclic monophosphate (8-pAcAmdPT-
	cGMP)
	Using general procedure A, 8-Br-cGMP was reacted with 4-acetamidothiophenol to give the
	title compound.
	Yield (Purity): 95% (>99%).
	HPLC: (35% MeOH, 10 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 275 nm (pH 7), ϵ = 21500 (est.).
	ESI-MS (+): m/z calculated for C₁₈H₂₀N₆O₈PS ([M + H]⁺): 511.08, found: 511.
	ESI-MS (−): m/z calculated for C₁₈H₁₈N₆O₈PS ([M − H]⁻): 509.06, found: 509.

83

	8-(4-Chlorophenylsulfonyl)guanosine-3′,5′-cyclic monophosphate (8-pCPS-cGMP)
	The title compound was synthesized from 8-pCPT-cGMP using general procedure O.
	Yield (Purity): 29% (>99%).
	HPLC: (19% MeCN, 10 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 241 (276, 311) nm (pH 7), ϵ = 21500 (est.).
	ESI-MS (+): m/z calculated for C₁₆H₁₆ClN₅O₉PS ([M + H]⁺): 520.00, found: 520.
	ESI-MS (−): m/z calculated for C₁₆H₁₄ClN₅O₉PS ([M − H]⁻): 517.99, found: 518.

84

	8-(4-Chlorophenylsulfoxide)-guanosine-3′,5′-cyclic monophosphate (8-(pCPS(O)-
	cGMP)
	The title compound was synthesized from 8-pCPT-cGMP using general procedure P.
	Yield (Purity): 27% (>99%).
	HPLC: (28% MeOH, 10 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 275 nm (pH 7), ϵ = 21500 (est.).
	ESI-MS (+): m/z calculated for C₁₆H₁₆ClN₅O₈PS ([M + H]⁺): 504.01, found: 504.
	ESI-MS (−): m/z calculated for C₁₆H₁₄ClN₅O₈PS ([M − H]⁻): 502.00, found: 502.

85

	8-((2-Ethoxyethyl)-4-thiophenylthio)guanosine-3′,5′-cyclic monophosphate (8-(2-
	EOE)-pTPT-cGMP)
	Using general procedure D, 8-pTPT-cGMP was reacted with 2-Bromoethyl ethyl ether to
	give the title compound.
	Yield (Purity): 64% (>99%).
	HPLC: (48% MeOH, 10 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 276 nm (pH 7), ϵ = 21500 (est.).
	ESI-MS (+): m/z calculated for C₂₀H₂₅N₅O₈PS ([M + H]⁺): 558.09, found: 558.
	ESI-MS (−): m/z calculated for C₂₀H₂₃N₅O₈PS₂([M − H]⁻): 556.07, found: 556.

86

	8-(4-Thiophenyl-4″-thiophenylthio)guanosine-3′,5′-cyclic monophosphate (8-pTP-
	pTPT-cGMP)
	Using general procedure B, 8-Br-cGMP was reacted with 4,4′-thiobisbenzenethiol to give
	the title compound.
	Yield (Purity): 13% (>99%).
	HPLC: (52% MeOH, 10 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 290 nm (pH 7), ϵ = 21500 (est.).
	ESI-MS (+): m/z calculated for C₂₂H₂₁N₅O₇PS₃([M + H]⁺): 594.03, found: 594.
	ESI-MS (−): m/z calculated for C₂₂H₁₉N₅O₇PS₃([M − H]⁻): 592.02, found: 592.

87

	8-(2-Azidoethylthio)guanosine-3′,5′-cyclic monophosphate (8-N₃-ET-cGMP)
	The title compound was synthesized from 8-T-cGMP and 1,2-dibromoethane using general
	procedure Q.
	Yield (Purity): 54% (>99%).
	HPLC: (9% MeCN, 30 mM NaH₂PO₄buffer, pH 6.8).
	UV-VIS: 275 nm (pH 7), ϵ = 13700 (est.).
	ESI-MS (+):(+): m/z calculated for C₁₂H₁₆N₈O₇PS ([M + H]⁺): 447.06, found: 447.
	ESI-MS (−): m/z calculated for C₁₂H₁₄N₈O₇PS ([M − H]⁻): 445.04, found: 445.

88

	8-(3-Aminopropyl)-(pentaethoxy)-methylamidomethylthio-guanosine-3′,5′-cyclic
	monophosphate (8-APr-(EO)₅-MAmdMT-cGMP)
	Using general procedure K, 8-CMT-cGMP (1 eq) was reacted with NH₂CH₂-PEG₅-
	(CH₂)₃NH₂(6 eq) to give the title compound.
	Yield (Purity): 39% (>95%).
	HPLC: (27% MeOH, 20 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 275 nm (pH 7), ϵ = 13700 (est.).
	ESI-MS (+): m/z calculated for C₂₆H₄₅N₇O₁₃PS ([M + H]⁺): 726.25, found: 726.
	ESI-MS (−): m/z calculated for C₂₆H₄₃N₇O₁₃PS ([M − H]⁻): 724.24, found 724.

89
	8-(2-Aminoethyl)-(octaethoxy)-amidomethylthioguanosine-3′,5′-cyclic
	monophosphate (8-AE-(EO)₈-AmdMT-cGMP)
	Using general procedure K, 8-CMT-cGMP (1 eq) was reacted with NH₂-PEG₈-(CH₂)₂NH₂
	(6 eq) to give the title compound.
	Yield (Purity): 53% (>99%).
	HPLC: (10% MeCN, 50 mM NaH₂PO₄buffer, pH 6.8).
	UV-VIS: λ_max= 275 nm (pH 7), ϵ = 13700 (est.).
	ESI-MS (+): m/z calculated for C₃₀H₅₃N₇O₁₆PS ([M + H]⁺): 830.30, found: 830.
	ESI-MS (−): m/z calculated for C₃₀H₅₁N₇O₁₆PS ([M − H]⁻): 828.29, found: 828.

90

	8-(2-Bromoethyl)-(pentaethoxy)-(4-thiophenylthio)guanosine-3′,5′-cyclic
	monophosphate (8-BrE-(EO)₅-pTPT-cGMP)
	Using general procedure E, 8-pTPT-cGMP was reacted with Br-PEG₅-CH₂CH₂Br. The title
	compound was isolated beside the dimeric analogue.
	Yield (Purity): 43% (>99%).
	HPLC: (52% MeOH, 10 mM TEAF buffer, pH 6.8).
	HPLC (analytical): (45% MeOH, 10 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 276 nm (pH 7), ϵ = 21500 (est.).
	ESI-MS (+): m/z calculated for C₂₈H₃₉BrN₅O₁₂PS₂Na ([M + Na]⁺): 834.09, found: 834.
	ESI-MS (−): m/z calculated for C₂₈H₃₈BrN₅O₁₂PS₂([M − H]⁻): 810.09, found: 810.

91

	8-(4-(Propargyloxy-(hexaethoxy)-methyl)-[1,2,3]-triazole-1-yl)-
	methylamidoethylthioguanosine-3′,5′-cyclic monophosphate (8-(4-(PargO-(EO)₆-
	Me)-[1,2,3]-Tz-1)-MAmdET-cGMP)
	Using general procedure S, 8-N3-MAmdET-cGMP (1 eq) was reacted with bis-propargyl-
	PEG₇(2 eq) to give the title compound. Conditions were chosen to additionally obtain the
	dimeric analogue (G 045).
	Yield (Purity): 38% (>99%).
	HPLC: (14% MeCN, 30 mM TEAF buffer, pH 6.8 then (after sepperation of dimer)
	15% MeCN, 10 mM TEAF buffer, pH 6.8).
	HPLC (analytical): (15% MeCN, 20 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 276 nm (pH 7), ϵ = 13700 (est.).
	ESI-MS (+): m/z calculated for C₃₂H₄₉N₉O₁₅PS ([M + Na]⁺): 862.28, found: 862.
	ESI-MS (−): m/z calculated for C₃₂H₄₇N₉O₁₅PS ([M − H]⁻): 860.27, found: 860.

92

	8-(4-Carboxyphenylthio)guanosine-3′,5′-cyclic monophosphate (8-pCarbT-cGMP)
	Using general procedure C, 8-Br-cGMP (1 eq) was reacted with 4-mercaptobenzoic acid
	(2 eq) at 60° C. in the absence of NaOH to give the title compound.
	Yield (Purity): 67% (>99%).
	HPLC: (9% MeCN, 20 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 277 nm (pH 7), ϵ = 21500 (est.).
	ESI-MS (−): m/z calculated for C₁₇H₁₅N₅O₉PS ([M − H]⁻): 496.03, found: 496.

93

	8-(4-Hydroxyphenylsulfonyl)-guanosine-3′,5′-cyclic monophosphate (8-pHPS-
	cGMP)
	The title compound was synthesized from 8-pHPT-cGMP using general procedure O.
	Yield (Purity): 38% (>99%).
	HPLC: (30% MeOH, 10 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 276 (310) nm (pH 7), ϵ = 21500 (est.).
	ESI-MS (+): m/z calculated for C₁₆H₁₇N₅O₁₀PS ([M + H]⁺): 502.04, found: 502.
	ESI-MS (−): m/z calculated for C₁₆H₁₅N₅O₁₀PS ([M − H]⁻): 500.03, found: 500.

94

	8-(4-Isopropylphenylsulfonyl)-guanosine-3′,5′-cyclic monophosphate (8-pIPrPS-
	cGMP)
	The title compound was synthesized from 8-pIPrPT-cGMP using general procedure O.
	Yield (Purity): 30% (>99%).
	HPLC: (40% MeOH, 10 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 276 (239, 306) nm (pH 7), ϵ = 21500 (est.).
	ESI-MS (+): m/z calculated for C₁₉H₂₃N₅O₉PS ([M + H]⁺): 528.10, found: 528.
	ESI-MS (−): m/z calculated for C₁₉H₂₁N₅O₉PS ([M − H]⁻): 526.08, found: 526.

95

	8-(4-Methylcarboxyphenylthio)-guanosine-3′,5′-cyclic monophosphate (8-
	pMeCarbPT-cGMP)
	Using general procedure A, 8-Br-cGMP (1 eq) was reacted with methyl 4-mercaptobenzoate
	(4 eq) at 60° C. replacing NaOH by borate buffer (100 mM, pH 10.4, ex) to give the title
	compound.
	Yield (Purity): 20% (>99%).
	HPLC: (22% MeCN, 50 mM NaH₂PO₄buffer, pH 6.7).
	UV-VIS: λ_max= 277 nm (pH 7), ϵ = 21500 (est.).
	ESI-MS (+): m/z calculated for C₁₈H₁₉N₅O₉PS ([M + H]⁺): 512.06, found: 512.
	ESI-MS (−): m/z calculated for C₁₈H₁₇N₅O₉PS ([M − H]⁻): 510.05, found: 510.

96

	8-Methylsulfonylguanosine-3′,5′-cyclic monophosphate (8-MSulf-cGMP)
	The title compound was synthesized from 8-MeS-cGMP using general procedure O.
	Yield (Purity): 35% (>99%).
	HPLC: (10% MeOH, 15 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 274 nm (pH 7), ϵ = 13700 (est.).
	ESI-MS (+): m/z calculated for C₁₁H₁₅N₅O₉PS ([M + H]⁺): 424.03, found: 424.
	ESI-MS (−): m/z calculated for C₁₁H₁₃N₅O₉PS ([M − H]⁻): 422.02, found: 422.

97

	8-(1-Bromo-2-naphthyl)methylthioguanosine-3′,5′-cyclic monophosphate (8-(1-Br-
	2-N)MT-cGMP)
	Using general procedure D, 8-T-cGMP was reacted with 1-bromo-2-
	bromomethylnaphthalene to give the title compound.
	Yield (Purity): 40% (>99%).
	HPLC: (27% MeCN, 50 mM NaH₂PO₄buffer, pH 6.7).
	UV-VIS: λ_max= 280 nm (pH 7), ϵ = 21500 (est.).
	ESI-MS (+): m/z calculated for C₂₁H₂₀BrN₅O₇PS ([M + H]⁺): 596.00, found: 596.
	ESI-MS (−): m/z calculated for C₂₁H₁₈BrN₅O₇PS ([M − H]⁻): 593.98, found: 594.

98

	8-(2-(1-Benzyl-[1,2,3]-triazole-4-yl)-ethylthio)guanosine-3′,5′-cyclic
	monophosphate (8-(1-Bn-[1,2,3]-Tz-4)-ET-cGMP)
	Using general procedure R, 8-Bu(3-yne)T-cGMP was reacted with benzyl azide to give the
	title compound.
	Yield (Purity): 30% (>98%).
	HPLC: (35% MeOH, 10 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 278 nm (pH 7), ϵ = 13700 (est.).
	ESI-MS (+): m/z calculated for C₂₁H₂₄N₈O₇PS ([M + H]⁺): 563.14, found: 563.
	ESI-MS (−): m/z calculated for C₂₁H₂₂N₈O₇PS ([M − H]⁻): 561.11, found: 561.

99

	8-(3-Fluoro-5-methoxybenzylthio)guanosine-3′,5′-cyclic monophosphate (8-(3-F-5-
	MeO)BT-cGMP)
	Using general procedure D, 8-T-cGMP was reacted with 3-fluoro-5-methoxybenzyl bromide
	to give the title compound.
	Yield (Purity): 40% (>99%).
	HPLC: (22% MeCN, 50 mM NaH₂PO₄buffer, pH 6.7).
	UV-VIS: λ_max= 278 nm (pH 7), ϵ = 21500 (est.).
	ESI-MS (+): m/z calculated for C₁₈H₂₀FN₅O₈PS ([M + H]⁺): 516.07, found: 516.
	ESI-MS (−): m/z calculated for C₁₈H₁₈FN₅O₈PS ([M − H]⁻): 514.06, found: 514.

100

	8-Pentafluorobenzylthioguanosine-3′,5′-cyclic monophosphate (8-PFBT-cGMP)
	Using general procedure D, 8-T-cGMP was reacted with 2,3,4,5,6-pentafluorobenzyl
	bromide to give the title compound.
	Yield (Purity): 35% (>99%).
	HPLC: (27% MeCN, 50 mM NaH₂PO₄buffer, pH 6.7).
	UV-VIS: λ_max= 278 nm (pH 7), ϵ = 21500 (est.).
	ESI-MS (+): m/z calculated for C₁₇H₁₄F₅N₅O₇PS ([M + H]⁺): 558.03, found: 558.
	ESI-MS (−): m/z calculated for C₁₇H₁₂F₅N₅O₇PS ([M − H]⁻): 556.01, found: 556.

101

	8-Triphenyliminophosphoranyl-guanosine-3′,5′-cyclic monophosphate (8-Ph₃PN-
	cGMP)
	The title compound was synthesized from 8-N₃-cGMP using general procedure W.
	Yield (Purity): 18% (>99%).
	HPLC: (26% MeCN, 20 mM NaH₂PO₄buffer, pH 6.8).
	UV-VIS: λ_max= 267 nm (pH 7), ϵ = 13700 (est.).
	ESI-MS (+): m/z calculated for C₂₈H₂₇N₆O₇P₂([M + H]⁺): 621.14, found: 621.
	ESI-MS (−): m/z calculated for C₂₈H₂₅N₆O₇P₂([M − H]⁻): 619.13, found: 619.

102

	8-(4-Chlorophenyl)guanosine-3′,5′-cyclic monophosphate (8-pCP-cGMP)
	Using general procedure X, 8-Br-cGMP was reacted with 4-chlorophenylboronic acid to give
	the title compound.
	Yield (Purity): 10% (>99%).
	HPLC: (17% MeCN, 10 mM TEAF buffer, pH 6.8).
	US-VIS: λ_max= 282 nm (pH 7), ϵ = 21500 (est.).
	ESI-MS (+): m/z calculated for C₁₆H₁₆ClN₅O₇P ([M + H]⁺): 456.05, found: 456.
	ESI-MS (−): m/z calculated for C₁₆H₁₄ClN₅O₇P ([M − H]⁻): 454.03, found: 454.

103

	8-(4-Fluorophenyl)guanosine-3′,5′-cyclic monophosphate (8-pFP-cGMP)
	Using general procedure X, 8-Br-cGMP was reacted with 4-fluorophenylboronic acid to give
	the title compound.
	Yield (Purity): 54% (>99%).
	HPLC: (31% MeOH, 10 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 280 nm (pH 7), ϵ = 21500 (est.).
	ESI-MS (+): m/z calculated for C₁₆H₁₆FN₅O₇P ([M + H]⁺): 440.08, found: 440.
	ESI-MS (−): m/z calculated for C₁₆H₁₄FN₅O₇P ([M − H]⁻): 438.06, found: 438.

104

	8-(2-Furyl)guanosine-3′,5′-cyclic monophosphate (8-(2-Fur)-cGMP)
	Using general procedure X, 8-Br-cGMP was reacted with 2-furylboronic acid to give the title
	compound.
	Yield (Purity): 40% (>99%).
	HPLC: (23% MeOH, 15 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 297 nm (pH 7), ϵ = 21500 (est.).
	ESI-MS (+): m/z calculated for C₁₄H₁₅N₅O₈P ([M + H]⁺): 412.07, found: 412.
	ESI-MS (−): m/z calculated for C₁₄H₁₃N₅O₈P ([M − H]⁻): 410.05, found: 410.

105

	8-(4-Hydroxyphenyl)guanosine-3′,5′-cyclic monophosphate (8-pHP-cGMP)
	Using general procedure X, 8-Br-cGMP was reacted with 4-hydroxyphenylboronic acid to
	give the title compound.
	Yield (Purity): 63% (>99%).
	HPLC: (21% MeOH, 10 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 282 nm (pH 7), ϵ = 21500 (est.).
	ESI-MS (+): m/z calculated for C₁₆H₁₇N₅O₈P ([M + H]⁺): 438.08, found: 438.
	ESI-MS (−): m/z calculated for C₁₆H₁₅N₅O₈P ([M − H]⁻): 436.07, found: 436.

106

	8-(4-Isopropylphenyl)guanosine-3′,5′-cyclic monophosphate (8-pIPrP-cGMP)
	Using general procedure X, 8-Br-cGMP was reacted with 4-isopropylphenylboronic acid to
	give the title compound.
	Yield (Purity): 33% (>99%).
	HPLC: (22% MeCN, 50 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 281 nm (ph 7), ϵ = 21500 (est.).
	ESI-MS (+): m/z calculated for C₁₉H₂₃N₅O₇P ([M + H]⁺): 464.13, found: 464.
	ESI-MS (−): m/z calculated for C₁₉H₂₁N₅O₇P ([M − H]⁻): 462.12, found: 462.

107

	8-Phenylguanosine-3′,5′-cyclic monophosphate (8-Phe-cGMP)
	Using general procedure X, 8-Br-cGMP was reacted with phenylboronic acid to give the title
	compound.
	2Yield (Purity): 79% (>99%).
	HPLC: (31% MeOH, 10 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 281 nm (pH 7), ϵ = 21500 (est.).
	ESI-MS (+): m/z calculated for C₁₆H₁₇N₅O₇P ([M + H]⁺): 422.09, found: 422.
	ESI-MS (−): m/z calculated for C₁₆H₁₅N₅O₇P ([M − H]⁻): 420.07, found: 420.

108

	β-Phenyl-1,N²-etheno-8-thioguanosine-3′,5′-cyclic monophosphate (PET-8-T-
	cGMP)
	Using modified general procedure C, NaSH (19 mM, 208 eq) in NaHCO₃-Buffer (pH 8.7)
	was added to 8-Br-PET-cGMP (19 mM, 1 eq) dissolved in NaHCO₃-buffer (pH 8.7) and
	reacted at 75° C. to give the title compound.
	Yield (Purity): 60% (>99%).
	HPLC: (21% MeCN, 50 mM NaH₂PO₄buffer, pH 6.7).
	UV-VIS: λ_max= 288 nm (pH 7), ϵ = 40000 (est.).
	ESI-MS (+): m/z calculated for C₁₈H₁₆N₅O₇PSNa ([M + Na]⁺): 500.04, found: 500.
	ESI-MS (−): m/z calculated for C₁₈H₁₅N₅O₇PS ([M − H]⁻): 476.04, found: 476.

109

	8-(2-Aminophenylthio)-β-phenyl-1,N²-ethenoguanosine-3′,5′-cyclic
	monophosphate (8-oAPT-PET-cGMP)
	Using general procedure A, 8-Br-PET-cGMP (1 eq) was reacted with 2-aminothiophenol
	(8 eq) in the presence of NaOH (2 eq) to give the title compound.
	Yield (Purity): 57% (>99%).
	HPLC: (22% MeCN, 50 mM NaH₂PO₄buffer, pH 6.7).
	UV-VIS: λ_max= 273 nm (pH 7), ϵ = 40000 (est.).
	ESI-MS (+): m/z calculated for C₂₄H₂₂N₆O₇PS ([M + H]⁺): 569.10, found: 569.
	ESI-MS (−): m/z calculated for C₂₄H₂₀N₆O₇PS ([M − H]⁻): 567.08, found: 567.

110

	8-Cyclohexylthio-β-phenyl-1,N²-ethenoguanosine-3′,5′-cyclic monophosphate (8-
	cHeT-PET-cGMP)
	Using general procedure A, 8-Br-PET-cGMP (1 eq) was reacted with Cyclohexanethiol
	(6.5 eq) in the presence of NaOH (2 eq) to give the title compound.
	Yield (Purity): 20% (>99%).
	HPLC: (22% MeCN, 50 mM NaH₂PO₄buffer, pH 6.7).
	UV-VIS: λ_max= 275 nm (pH 7), ϵ = 40000 (est.).
	ESI-MS (+): m/z calculated for C₂₄H₂₇N₅O₇PS ([M + H]⁺): 560.14, found: 560.
	ESI-MS (−): m/z calculated for C₂₄H₂₅N₅O₇PS ([M − H]⁻): 558.12, found: 558.

111

	8-Cyclopentylthio-β-phenyl-1,N²-ethenoguanosine-3′,5′-cyclic monophosphate
	(8-cPeT-PET-cGMP)
	Using general procedure A, 8-Br-PET-cGMP (1 eq) was reacted with cyclopentanethiol
	(8 eq) in the presence of NaOH (2 eq) to give the title compound.
	Yield (Purity): 20% (>99%).
	HPLC: (22% MeCN, 50 mM NaH2PO4 buffer, pH 6.7).
	UV-VIS: λ_max= 276 nm (pH 7), ϵ = 40000 (est.).
	ESI-MS (+): m/z calculated for C₂₃H₂₅O₇PS ([M + H]⁺): 546.12, found: 546.
	ESI-MS (−): m/z calculated for C₂₃H₂₃N₅O₇PS ([M − H]⁻): 544.11, found: 544.

112

	8-(4-Methylphenylthio)-β-phenyl-1,N²-ethenoguanosine-3′,5′-cyclic
	monophosphate (8-pMePT-PET-cGMP)
	Using general procedure A, 8-Br-PET-cGMP (1 eq) was reacted with p-toluenethiol (8 eq) in
	the presence of NaOH (2 eq) to give the title compound.
	Yield (Purity): 70% (>99%).
	HPLC: (22% MeCN, 50 mM NaH₂PO₄, pH 6.7).
	UV-VIS: λ_max= 275 nm (pH 7), ϵ = 40000 (est.).
	ESI-MS (+): m/z calculated for C₂₅H₂₃N₅O₇PS ([M + H]⁺): 568.11, found: 568.
	ESI-MS (−): m/z calculated for C₂₅H₂₁N₅O₇PS ([M − H]⁻): 566.09, found: 566.

113

	8-(4-Methoxyphenylthio)-β-phenyl-1,N²-ethenoguanosine-3′,5′-cyclic
	monophosphate (8-pMeOPT-PET-cGMP)
	Using general procedure A, 8-Br-PET-cGMP (1 eq) was reacted with 4-
	methoxybenzenethiol (8 eq) in the presence of NaOH (2 eq) to give the title compound.
	Yield (Purity): 82% (>99%).
	HPLC: (22% MeCN, 50 mM NaH₂PO₄buffer, pH 6.7).
	UV-VIS: λ_max= 274 nm (pH 7), ϵ = 40000 (est.).
	ESI-MS (+): m/z calculated for C₂₅H₂₃N₅O₈PS ([M + H]⁺): 584.10, found: 584.
	ESI-MS (−): m/z calculated for C₂₅H₂₁N₅O₈PS ([M − H]⁻): 582.08, found: 582.

114

	8-(3-(2-Methyl)fuanyl)thio-β-phenyl-1,N²-ethenoguanosine-3′,5′-cyclic
	monophosphate (8-(3-(2-Me)-FU)T-PET-cGMP)
	Using general procedure A, 8-Br-PET-cGMP (1 eq) was reacted with 2-methyl-3-furanethiol
	(6 eq) in the presence of NaOH (2 eq) to give the title compound.
	Yield (Purity): 70% (>99%).
	HPLC: (22% MeCN, 50 mM NaH₂PO₄buffer, pH 6.7).
	UV-VIS: λ_max= 273 nm (pH 7), ϵ = 40000 (est.).
	ESI-MS (+): m/z calculated for C₂₃H₂₁N₅O₈PS ([M + H]⁺): 558.08, found: 558.
	ESI-MS (−): m/z calculated for C₂₃H₁₉N₅O₈PS ([M − H]⁻): 556.07, found: 556.

115

	8-(7-(4-Methyl)coumarinyl)thio-β-phenyl-1,N²-ethenoguanosine-3′,5′-cyclic
	monophosphate (8-(7-(4-Me)-Cou)T-PET-cGMP)
	Using general procedure A, 8-Br-PET-cGMP (1 eq) was reacted with 7-mercapto-4-
	methylcoumarin (6 eq) in the presence of NaOH (2 eq) to give the title compound.
	Yield (Purity): 80% (>99%).
	HPLC: (22% MeCN, 50 mM NaH₂PO₄buffer, pH 6.7).
	UV-VIS: λ_max= 277 nm (pH 7), ϵ = 40000 (est.).
	ESI-MS (+): m/z calculated for C₂₈H₂₃N₅O₉PS ([M + H]⁺): 636.09, found: 636.
	ESI-MS (−): m/z calculated for C₂₈H₂₁N₅O₉PS ([M − H]⁻): 634.08, found: 634.

116
	8-(2-Naphthyl)thio-β-phenyl-1,N²-ethenoguanosine-3′,5′-cyclic monophosphate
	(8-(2-N)T-PET-cGMP)
	Using general procedure A, 8-Br-PET-cGMP (1 eq) was reacted with 2-naphthalenethiol
	(6 eq) in the presence of NaOH (2 eq) to give the title compound.
	Yield (Purity): 30% (>99%).
	HPLC: (22% MeCN, 50 mM NaH₂PO₄buffer, pH 6.7).
	UV-VIS: λ_max= 275 nm (pH 7), ϵ = 40000 (est.).
	ESI-MS (+): m/z calculated for C₂₈H₂₃N₅O₇PS ([M + H]⁺): 604.11, found: 604.
	ESI-MS (−): m/z calculated for C₂₈H₂₁N₅O₇PS ([M − H]⁻): 602.09, found: 602.

117

	β-Phenyl-1,N²-etheno-8-(2-thiophenyl)thioguanosine-3′,5′-cyclic monophosphate
	(PET-8-(2-Tp)T-cGMP)
	Using general procedure A, 8-Br-PET-cGMP (1 eq) was reacted with 2-thiophenethiol (6 eq)
	in the presence of NaOH (2 eq) to give the title compound.
	Yield (Purity): 60% (>99%).
	HPLC: (22% MeCN, 50 mM NaH₂PO₄buffer, pH 6.7).
	UV-VIS: λ_max= 275 nm (pH 7), ϵ = 40000 (est.).
	ESI-MS (+): m/z calculated for C₂₂H₁₉N₅O₇PS₂([M + H]⁺): 560.05, found: 560.
	ESI-MS (−): m/z calculated for C₂₂H₁₇N₅O₇PS₂([M − H]⁻): 558.03, found: 558.

118

	β-Phenyl-1,N²-etheno-8-(2-phenylethyl)thioguanosine-3′,5′-cyclic
	monophosphate (PET-8-PhEtT-cGMP)
	Using general procedure A, 8-Br-PET-cGMP (1 eq) was reacted with 2-phenylethanethiol
	(6 eq) in the presence of NaOH (2 eq) to give the title compound.
	Yield (Purity): % (>99%).
	HPLC: (22% MeCN, 50 mM NaH₂PO₄buffer, pH 6.7).
	UV-VIS: λ_max= 276 nm (pH 7), ϵ = 40000 (est.).
	ESI-MS (+): m/z calculated for C₂₆H₂₅N₅O₇PS ([M + H]⁺): 581.12, found: 582.
	ESI-MS (−): m/z calculated for C₂₆H₂₃N₅O₇PS ([M − H]⁻): 580.11, found: 580.

119

	8-Carboxymethylthio-β-phenyl-1,N²-ethenoguanosine-3′,5′-cyclic
	monophosphate (8-CMT-PET-cGMP)
	Using general procedure C, 8-Br-PET-cGMP was reacted with mercaptoacetic acid to give
	the title compound.
	Yield (Purity): 68% (>99%).
	HPLC: (14% MeCN, 50 mM NaH₂PO₄buffer, pH 6.8).
	UV-VIS: λ_max= 273 nm (pH 7), ϵ = 40000 (est.).
	ESI-MS (+): m/z calculated for C₂₀H₁₉N₅O₉PS ([M + H]⁺): 536.06, found: 536.
	ESI-MS (−): m/z calculated for C₂₀H₁₇N₅O₉PS ([M − H]⁻): 534.05, found: 534.

120

	8-Amidomethylthio-β-phenyl-1,N²-ethenoguanosine-3′,5′-cyclic monophosphate
	(8-AmdMT-PET-cGMP)
	In a one pot synthesis 8-T-cGMP was reacted with 2-bromoacetamide following general
	procedure D to give 8-AmdMT-cGMP, which was transformed to the title compound by
	applying general procedure Y.
	Yield (Purity): 19% (>99%).
	HPLC: (35% MeOH, 10 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 273 nm (pH 7), ϵ = 40000 (est.).
	ESI-MS (+): m/z calculated for C₂₀H₂₀N₆O₈PS ([M + H]⁺): 535.08, found: 535.
	ESI-MS (−): m/z calculated for C₂₀H₁₈N₆O₈PS ([M − H]⁻): 533.06, found: 533.

121

	8-(4-Boronatephenylthio)-β-phenyl-1,N²-ethenoguanosine-3′,5′-cyclic
	monophosphate (8-(pB(OH)₂P)T-PET-cGMP)
	Using general procedure A, 8-Br-PET-cGMP was reacted with 4-mercaptophenylboronic
	acid to give the title compound.
	Yield (Purity): 25% (>99%).
	HPLC: (52% MeOH, 10 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 276 nm (pH 7), ϵ = 40000 (est.).
	ESI-MS (+): m/z calculated for C₂₄H₂₂BN₅O₉PS ([M + H]⁺): 598.10, found: 598.
	ESI-MS (−): m/z calculated for C₂₄H₂₀BN₅O₉PS ([M − H]⁻): 596.08, found: 596.

122

	8-Ethylthio-β-phenyl-1,N2-ethenoguanosine-3′,5′-cyclic monophosphate (8-ET-
	PET-cGMP)
	Using general procedure A, 8-Br-PET-cGMP (1 eq) was reacted with ethanethiol (12 eq) in
	a tube with screw cap at 70° C. to give the title compound.
	Yield (Purity): 46% (>99%).
	HPLC: (15% MeCN, 50 mM NaH₂PO₄buffer, pH 6.7).
	UV-VIS: λ_max= 274 nm (pH 7), ϵ = 40000 (est.).
	ESI-MS (+): m/z calculated for C₂₀H₂₁N₅O₇PS ([M + H]⁺): 506.09, found: 506.
	ESI-MS (−): m/z calculated for C₂₀H₁₉N₅O₇PS ([M − H]⁻): 504.07, found: 504.

123

	8-(4-Fluorophenylthio)-β-phenyl-1,N²-ethenoguanosine-3′,5′-cyclic
	monophosphate (8-pFPT-PET-cGMP)
	Using general procedure A, 8-Br-PET-cGMP (1 eq) was reacted with 4-fluorothiophenol
	(4 eq) in the presence of NaOH (2 eq) to give the title compound.
	Yield (Purity): 36% (>99%).
	HPLC: (22% MeCN, 50 mM NaH₂PO₄buffer, pH 6.7).
	UV-VIS: λ_max= 276 nm (pH 7), ϵ = 40000 (est.).
	ESI-MS (+): m/z calculated for C₂₄H₂₀FN₅O₇PS ([M + H]⁺): 572.08, found: 572.
	ESI-MS (−): m/z calculated for C₂₄H₁₈FN₅O₇PS ([M − H]⁻): 570.06, found: 570.

124

	8-Methylthio-β-phenyl-1,N²-ethenoguanosine-3′,5′-cyclic monophosphate (8-
	MeS-PET-cGMP)
	Using general procedure A, 8-Br-PET-cGMP (1 eq) was reacted with sodium
	methanethiolate (4 eq) to give the title compound.
	Yield (Purity): 75% (>99%).
	HPLC: (15% MeCN, 50 mM NaH₂PO₄buffer, pH 6.7).
	UV-VIS: λ_max= 272 nm (pH 7), ϵ = 40000 (est.).
	ESI-MS (+): m/z calculated for C₁₉H₁₉N₅O₇PS ([M + H]⁺): 492.07, found: 492.
	ESI-MS (−): m/z calculated for C₁₉H₁₇N₅O₇PS ([M − H]⁻): 490.06, found: 490.

125

	β-Phenyl-1,N²-etheno-8-propylthio-guanosine-3′,5′-cyclic monophosphate (PET-
	8-PrT-cGMP)
	Using general procedure A, 8-Br-PET-cGMP (1 eq) was reacted with propanethiol (16 eq) in
	a tube with screw cap at 70° C. to give the title compound.
	Yield (Purity): 46% (>99%).
	HPLC: (15% MeCN, 50 mM NaH₂PO₄buffer, pH 6.7).
	UV-VIS: λ_max= 273 nm (pH 7), ϵ = 40000 (est.).
	ESI-MS (+): m/z calculated for C₂₁H₂₃N₅O₇PS ([M + H]⁺): 520.11, found: 521.
	ESI-MS (−): m/z calculated for C₂₁H₂₁N₅O₇PS ([M − H]⁻): 518.09, found: 518.

126

	8-Azidoethylthio-β-phenyl-1,N²-ethenoguanosine-3′,5′-cyclic monophosphate (8-
	N₃-ET-PET-cGMP)
	The title compound was synthesized from PET-8-T-cGMP and 1,2-dibromoethane using
	general procedure Q.
	Yield (Purity): 41% (>99%).
	HPLC: (23% MeCN, 40 mM NaH₂PO₄buffer, pH 6.8).
	UV-VIS: λ_max= 273 nm (pH 7), ϵ = 40000 (est.).
	ESI-MS (+): m/z calculated for C₂₀H₂₀N₈O₇PS ([M + H]⁺): 547.09, found: 547.
	ESI-MS (−): m/z calculated for C₂₀H₁₈N₈O₇PS ([M − H]⁻): 545.08, found: 545.

127

	β-Phenyl-1,N²-etheno-8-(4-trifluoromethylphenylthio)guanosine-3′,5′-cyclic
	monophosphate (PET-8-pTFMePT-cGMP)
	Using general procedure A, 8-Br-PET-cGMP (1 eq) was reacted with
	4-(trifluoromethyl)thiophenol (4 eq) in the presence of NaOH (2 eq) to give the title
	compound.
	Yield (Purity): 60% (>99%).
	HPLC: (22% MeCN, 50 mM NaH₂PO₄buffer, pH 6.7).
	UV-VIS: λ_max= 276 nm (pH 7), ϵ = 40000 (est.).
	ESI-MS (+): m/z calculated for C₂₅H₂₀F₃N₅O₇PS ([M + H]⁺): 622.08, found: 622.
	ESI-MS (−): m/z calculated for C₂₅H₁₈F₃N₅O₇PS ([M − H]⁻): 620.06, found: 620.

128

	8-(4-Chlorophenylsulfonyl)-β-phenyl-1,N²-ethenoguanosine-3′,5′-cyclic
	monophosphate (8-pCPS-PET-cGMP)
	Using general procedure Y, 8-pCPS-cGMP was reacted with 2-bromoacetophenone to give
	the title compound.
	Yield (Purity): 45% (>99%).
	HPLC: (30% MeCN, 10 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 287 nm (pH 7), ϵ = 40000 (est.).
	ESI-MS (+): m/z calculated for C₂₄H₂₀ClN₅O₉PS ([M + H]⁺): 620.04, found: 620.
	ESI-MS (−): m/z calculated for C₂₄H₁₈ClN₅O₉PS ([M − H]⁻): 618.03, found: 618.

129

	8-(4-Isopropylphenylthio)-β-phenyl-1,N²-ethenoguanosine-3′,5′-cyclic
	monophosphate (8-pIPrPT-PET-cGMP)
	Using general procedure A, 8-Br-PET-cGMP (1 eq) was reacted with 4-isopropylthiophenol
	(4 eq) in the presence of NaOH (2 eq) at 60° C. to give the title compound.
	Yield (Purity): 20% (>99%).
	HPLC: (27% MeCN, 50 mM NaH₂PO₄buffer, pH 6.8).
	UV-VIS: λ_max= 274 nm (pH 7), ϵ = 40000 (est.).
	ESI-MS (+): m/z calculated for C₂₇H₂₇N₅O₇PS ([M + H]⁺): 596.14, found: 596.
	ESI-MS (−): m/z calculated for C₂₇H₂₅N₅O₇PS ([M − H]⁻): 594.12, found: 594.

130

	8-(4-Isopropylphenylsulfonyl)-β-phenyl-1,N²-ethenoguanosine-3′,5′-cyclic
	monophosphate (8-pIPrPS-PET-cGMP)
	Using general procedure Y, 8-pIPrPS-cGMP was reacted with 2-bromoacetophenone to
	give the title compound.
	Yield (Purity): 38% (>99%).
	HPLC: (61% MeOH, 10 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 285 nm (pH 7), ϵ = 40000 (est.).
	ESI-MS (+): m/z calculated for C₂₇H₂₇N₅O₉PS ([M + H]⁺): 628.13, found: 628.
	ESI-MS (−): m/z calculated for C₂₇H₂₅N₅O₉PS ([M − H]⁻): 626.11, found: 626.

131

	8-(4-Chlorophenyl)-β-phenyl-1,N²-ethenoguanosine-3′,5′-cyclic monophosphate
	(8-pCP-PET-cGMP)
	Using general procedure X, 8-Br-PET-cGMP was reacted with 4-chlorphenylboronic acid to
	give the title compound.
	Yield (Purity): 29% (>99%).
	HPLC: (27% MeCN, 25 mM NaH₂PO₄buffer, pH 6.8).
	UV-VIS: λ_max= 279 nm (pH 7), ϵ = 40000 (est.).
	ESI-MS (+): m/z calculated for C₂₄H₂₀ClN₅O₇P ([M + H]⁺): 556.08, found: 556.
	ESI-MS (−): m/z calculated for C₂₄H₁₈ClN₅O₇P ([M − H]⁻): 554.06, found: 554.

132

	8-(4-Hydroxyphenyl)-β-phenyl-1,N²-ethenoguanosine-3′,5′-cyclic
	monophosphate (8-pHP-PET-cGMP)
	Using general procedure X, 8-Br-PET-cGMP was reacted with 4-hydroxyphenylboronic acid
	to give the title compound.
	Yield (Purity): 43% (>99%).
	HPLC: (20% MeCN, 25 mM NaH₂PO₄buffer, pH 6.8).
	UV-VIS: λ_max= 279 nm (pH 7), ϵ = 40000 (est.).
	ESI-MS (+): m/z calculated for C₂₄H₂₁N₅O₈P ([M + H]⁺): 538.11, found: 538.
	ESI-MS (−): m/z calculated for C₂₄H₁₉N₅O₈P ([M − H]⁻): 536.10, found: 536.

133

	8-(4-Isopropylphenyl)-β-phenyl-1,N²-ethenoguanosine-3′,5′-cyclic
	monophosphate (8-pIPrP-PET-cGMP)
	Using general procedure X, 8-Br-PET-cGMP was reacted with 4-isopropylphenylboronic
	acid to give the title compound.
	Yield (Purity): 25% (>99%).
	HPLC: (31% MeCN, 30 mM NaH₂PO₄buffer, pH 6.8).
	UV-VIS: λ_max= 277 nm (pH 7), ϵ = 40000 (est.).
	ESI-MS (+): m/z calculated for C₂₇H₂₇N₅O₇P ([M + H]⁺): 564.16, found: 564.
	ESI-MS (−): m/z calculated for C₂₇H₂₅N₅O₇P ([M − H]⁻): 562.15, found: 562.

134

	8-Bromo-(4-methoxy-β-phenyl-1,N2-etheno)guanosine-3′,5′-cyclic
	monophosphate (8-Br-pMeO-PET-cGMP)
	Using general procedure Y, 8-Br-cGMP was reacted with 2-bromo-4′-methoxyacetophenone
	to give the title compound.
	Yield (Purity): 17% (>97%).
	HPLC: (50% MeOH, 25 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 267 nm (pH 7), ϵ = 40000 (est.).
	ESI-MS (+): m/z calculated for C₁₉H₁₇BrN₅O₈PNa ([M + Na]⁺): 575.99, found: 576.
	ESI-MS (−): m/z calculated for C₁₉H₁₆BrN₅O₈P ([M − H]⁻): 551.99, found: 552.

135

	8-Bromo-(4-methyl-β-phenyl-1,N2-etheno)guanosine-3′,5′-cyclic
	monophosphate (8-Br-pMe-PET-cGMP)
	Using general procedure Y, 8-Br-cGMP was reacted with 2-bromo-4′-methylacetophenone
	to give the title compound.
	Yield (Purity): 11% (>98%).
	HPLC: (23% i-PrOH, 50 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 262 nm (pH 7), ϵ = 40000 (est.).
	ESI-MS (+): m/z calculated for C₁₉H₁₇BrN₅O₇PNa ([M + Na]⁺): 559.99, found: 560.
	ESI-MS (−): m/z calculated for C₁₉H₁₆BrN₅O₇P ([M − H]⁻): 536.00, found: 536.

136

	alpha-Benzoyl-beta-phenyl-1,N2-etheno-8-bromoguanosine-3′,5′-cyclic
	monophosphate (Bnz-PET-8-Br-cGMP)
	Using general procedure Y, 8-Br-cGMP was reacted with 2-bromo-1,3-diphenylpropane-1,3-
	dione to give the title compound.
	Yield (Purity): 12% (>94%).
	HPLC: (45% MeOH, 25 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 255 nm (pH 7), ϵ = 40000 (est.).
	ESI-MS (+): m/z calculated for C₂₅H₁₉BrN₅O₈PNa ([M + Na]⁺): 650.01, found: 650.
	ESI-MS (−): m/z calculated for C₂₅H₁₈BrN₅O₈P ([M − H]⁻): 626.01, found: 626.

137

	8-Bromo-(4-chloro-β-phenyl-1,N2-etheno)guanosine-3′,5′-cyclic
	monophosphate (8-Br-pCl-PET-cGMP)
	Using general procedure Y, 8-Br-cGMP was reacted with 2-bromo-4′-chloroacetophenone
	to give the title compound.
	Yield (Purity): 28% (>99%).
	HPLC: (57% MeOH, 20 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 263 nm (pH 7), ϵ = 40000 (est.).
	ESI-MS (+): m/z calculated for C₁₈H₁₃BrClN₅O₇PNa₂([M + 2Na − H]⁺): 601.92, found: 602.
	ESI-MS (−): m/z calculated for C₁₈H₁₃BrClN₅O₇P ([M − H]⁻): 555.94, found: 556.

138

	8-Bromo-(3-nitro-β-phenyl-1,N2-etheno)guanosine-3′,5′-cyclic monophosphate
	(8-Br-mN-PET-cGMP)
	Using general procedure Y, 8-Br-cGMP was reacted with 2-bromo-3′-nitroacetophenone to
	give the title compound.
	Yield (Purity): 14% (>98%).
	HPLC: (45% MeOH, 25 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 261 nm (pH 7), ϵ = 40000 (est.).
	ESI-MS (+): m/z calculated for C₁₈H₁₅BrN₆O₉P ([M + H]⁺): 568.98, found: 569.
	ESI-MS (−): m/z calculated for C₁₈H₁₃BrN₆O₉P ([M − H]⁻): 566.97, found: 567.

139

	8-Bromo-(β-tert.-butyl-1,N2-etheno)guanosine-3′,5′-cyclic monophosphate (8-
	Br-tBuET-cGMP)
	Using general procedure Y, 8-Br-cGMP was reacted with 1-bromo-3,3-dimethyl-2-butanone
	to give the title compound.
	Yield (Purity): 38% (>99%).
	HPLC: (48% MeOH, 10 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= nm (pH 7), ϵ = 40000 (est.).
	ESI-MS (+): m/z calculated for C₁₆H₂₀BrN₅O₇P ([M + H]⁺): 504.03, found: 504.
	ESI-MS (−): m/z calculated for C₁₆H₁₈BrN₅O₇P ([M − H]⁻): 502.01, found: 502.

140

	8-Bromo-(2-methoxy-β-phenyl-1,N2-etheno)guanosine-3′,5′-cyclic
	monophosphate (8-Br-oMeO-PET-cGMP)
	Using general procedure Y, 8-Br-cGMP was reacted with 2-bromo-2′-methoxyacetophenone
	to give the title compound.
	Yield (Purity): 4% (>99%).
	HPLC: (52% MeOH, 10 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 256 nm (pH 7), ϵ = 40000 (est.).
	ESI-MS (+): m/z calculated for C₁₉H₁₈BrN₅O8P ([M + H]⁺): 554.01, found: 554.
	ESI-MS (−): m/z calculated for C₁₉H₁₆BrN₅O8P ([M − H]⁻): 551.99, found: 552.

141

	8-Bromo-(3-methoxy-β-phenyl-1,N2-etheno)guanosine-3′,5′-cyclic
	monophosphate (8-Br-mMe-PET-cGMP)
	Using general procedure Y, 8-Br-cGMP was reacted with 2-bromo-3′-methoxyacetophenone
	to give the title compound.
	Yield (Purity): 10% (>99%).
	HPLC: (23% i-PrOH, 50 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 258 nm (pH 7), ϵ = 40000 (est.).
	ESI-MS (+): m/z calculated for C₁₉H₁₈BrN₅O8P ([M + H]⁺): 554.01, found: 554.
	ESI-MS (−): m/z calculated for C₁₉H₁₆BrN₅O8P ([M − H]⁻): 551.99, found: 552.

142

	8-Bromo-(2,4-dimethoxy-β-phenyl-1,N2-etheno)guanosine-3′,5′-cyclic
	monophosphate (8-Br-o,pDMeO-PET-cGMP)
	Using general procedure Y, 8-Br-cGMP was reacted with 2-bromo-2′,4′-
	dimethoxyacetophenone to give the title compound.
	Yield (Purity): 14% (>98%).
	HPLC: (18% MeCN, 50 mM NaH₂PO₄buffer, pH 6.7).
	UV-VIS: λ_max= 267 nm (pH 7), ϵ = 40000 (est.).
	ESI-MS (+): m/z calculated for C₂₀H₂₀BrN₅O₉P ([M + H]⁺): 584.02, found: 584.
	ESI-MS (−): m/z calculated for C₂₀H₁₈BrN₅O₉P ([M − H]⁻): 582.00, found: 582.

143

	8-Bromo-(4-pyridinyl-1,N2-etheno)guanosine-3′,5′-cyclic monophosphate (8-Br-
	(4-Pyr)ET-cGMP)
	Using general procedure Y, 8-Br-cGMP was reacted with 2-bromo-1-(4-pyridinyl)-1-
	ethanone hydrochloride to give the title compound.
	Yield (Purity): 31% (>98%).
	HPLC: (15% MeCN, 50 mM NaH₂PO₄buffer, pH 6.7).
	UV-VIS: λ_max= 266 nm (pH 7), ϵ = 40000 (est.).
	ESI-MS (+): m/z calculated for C₁₇H₁₅BrN₆O₇P ([M + H]⁺): 524.99, found: 525.
	ESI-MS (−): m/z calculated for C₁₇H₁₃BrN₆O₇P ([M − H]⁻): 522.98, found: 523.

144

	8-Bromo-(3-thiophen-yl-1,N2-etheno)guanosine-3′,5′-cyclic monophosphate (8-
	Br-(3-Tp)ET-cGMP)
	Using general procedure Y, 8-Br-cGMP was reacted with 3-(bromoacetyl)-thiophene to give
	the title compound.
	Yield (Purity): 52% (>99%).
	HPLC: (15% MeCN, 50 mM NaH₂PO₄buffer, pH 6.7).
	UV-VIS: λ_max= 261 nm (pH 7), ϵ = 40000 (est.).
	ESI-MS (+): m/z calculated for C₁₆H₁₄BrN₅O₇PS ([M + H]⁺): 529.95, found: 530.
	ESI-MS (−): m/z calculated for C₁₆H₁₂BrN₅O₇PS ([M − H]⁻): 527.94, found: 528.

145

	8-Bromo-(4-fluoro-β-phenyl-1,N2-etheno)guanosine-3′,5′-cyclic
	monophosphate (8-Br-pF-PET-cGMP)
	Using general procedure Y, 8-Br-cGMP was reacted with 4-fluorophenacyl bromide to give
	the title compound.
	Yield (Purity): 25% (>99%).
	HPLC: (15% MeCN, 50 mM NaH₂PO₄buffer, pH 6.7).
	UV-VIS: λmax = 257 nm (pH 7), ϵ = 40000 (est.).
	ESI-MS (+): m/z calculated for C₁₈H₁₅BrFN₅O₇P ([M + H]⁺): 541.99, found: 542.
	ESI-MS (−): m/z calculated for C₁₈H₁₃BrFN₅O₇P ([M − H]⁻): 539.97, found: 540.

146

	8-Bromo-1,N2-ethenoguanosine-3′,5′-cyclic monophosphate (8-Br-ET-cGMP)
	Using general procedure Y, 8-Br-cGMP was reacted with chloroacetaldehyde to give the
	title compound.
	Yield (Purity): 10% (>99%).
	HPLC: (9% MeCN, 20 mM TEAF buffer, pH 6.9).
	UV-VIS: λmax = 287 nm (pH 7), ϵ = 14600 (est.).
	ESI-MS (+): m/z calculated for C₁₂H₁₂BrN₅O₇P ([M + H]⁺): 447.97, found: 448.
	ESI-MS (−): m/z calculated for C₁₂H₁₀BrN₅O₇P ([M − H]⁻): 445.95, found: 446.

147

	8-Bromo-(3-hydroxy-β-phenyl-1,N2-etheno)guanosine-3′,5′-cyclic
	monophosphate (8-Br-mH-PET-cGMP)
	Using general procedure Y, 8-Br-cGMP was reacted with 3′-benzoyloxy-2-
	bromoacetophenone to give the O-benzoyl protected analogue of the title compound. Prior
	to chromatographic workup the crude product was subjected to NaOH (2M, 10 eq), to effect
	cleavage of the protecting group, and neutralized with HCl (1M).
	Yield (Purity): 13% (>99%).
	HPLC: (43% MeOH, 10 mM TEAF buffer, pH 6.8), second chromatographic workup
	(20% MeCN, 10 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 258 nm (pH 7), ϵ= 40000 (est.).
	ESI-MS (+): m/z calculated for C₁₈H₁₆BrN₅O₈P ([M + H]⁺): 539.99, found: 540.
	ESI-MS (−): m/z calculated for C₁₈H₁₄BrN₅O₈P ([M − H]⁻): 537.98, found: 538.

148

	8-Bromo-(4-hydroxy-β-phenyl-1,N²-etheno)guanosine-3′,5′-cyclic
	monophosphate (8-Br-pHPET-cGMP)
	Using general Y, 8-Br-cGMP was reacted with 4-(bromoacetyl)-phenyl benzoate
	to give the O-benzoyl protected analogue of the title compound. Prior to chromatographic
	workup the crude product was subjected to NaOH (2M, 10 eq), to effect cleavage of the
	protecting group, and neutralized with HCl (1M).
	Yield (Purity): 12% (>99%).
	HPLC: (43% MeOH, 10 mM TEAF buffer, pH 6.8), second chromatographic
	workup (20% MeCN, 10 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 267 nm (pH 7), ϵ = 40000 (est.).
	ESI-MS (+): m/z calculated for C₁₈H₁₆BrN₅O₈P ([M + H]⁺): 539.99, found: 540.
	ESI-MS (−): m/z calculated for C₁₈H₁₄BrN₅O₈P ([M − H]⁻): 537.98, found: 538.

149

	8-Bromo-(β-(2,3-dihydro-1,4-benzodioxin)-1,N2-etheno)guanosine-3′,5′-cyclic
	monophosphate (8-Br-(2,3-DHy-1,4-BnzDiox)-ET-cGMP)
	Using general procedure Y, 8-Br-cGMP was reacted with 2-bromo-1-(2,3-dihydro-1,4-
	benzodioxin-6-yl)ethanone to give the title compound.
	Yield (Purity): 8% (>99%).
	HPLC: (52% MeOH, 10 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 267 nm (pH 7), ϵ = 40000 (est.).
	ESI-MS (+): m/z calculated for C₂₀H₁₈BrN₅O₉P ([M + H]⁺): 582.00, found: 582.
	ESI-MS (−): m/z calculated for C₂₀H₁₆BrN₅O₉P ([M − H]⁻): 579.99, found: 580.

150

	8-Bromo-(4-methylsulfonamido-β-phenyl-1,N2-etheno)guanosine-3′,5′-cyclic
	monophosphate (8-Br-pMSulfAmd-PET-cGMP)
	Using general procedure Y, 8-Br-cGMP was reacted with N-[4-(2-bromoacetyl)phenyl]-
	methanesulfonamide to give the title compound.
	Yield (Purity): 25% (>97%).
	HPLC: (45% MeOH, 10 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 270 nm (pH 7), ϵ = 40000 (est.).
	ESI-MS (+): m/z calculated for C₁₉H₁₉BrN₆O₉PS ([M + H]⁺): 616.99, found: 617.
	ESI-MS (+): m/z calculated for C₁₉H₁₇BrN₆O₉PS ([M − H]⁻): 614.97, found: 615.

151

	8-Bromo-(4-cyano-β-phenyl-1,N²-etheno)guanosine-3′,5′-cyclic monophosphate
	(8-Br-pCN-PET-cGMP)
	Using general procedure Y, 8-Br-cGMP was reacted with 2-bromo-4′-cyanoacetophenone to
	give the title compound.
	Yield (Purity): 12% (>99%).
	HPLC: (45% MeOH, 10 mM TEAF buffer, pH 6.8), second chromatographic workup
	(22% MeCN, 10 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 272 nm (pH 7), ϵ = 40000 (est.).
	ESI-MS (+): m/z calculated for C₁₉H₁₅BrN₆O₇P ([M + H]⁺): 548.99, found: 549.
	ESI-MS (−): m/z calculated for C₁₉H₁₃BrN₆O₇P ([M − H]⁻): 546.98, found: 547.

152

	8-Bromo-(α-phenyl-β-methyl-1,N²-etheno)guanosine-3′,5′-cyclic
	monophosphate (8-Br-alpha-Phe-beta-Me-ET-cGMP)
	Using general procedure Y, 8-Br-cGMP was reacted with 1-bromo-1-phenylpropan-2-one to
	give the title compound.
	Yield (Purity): 9% (>99%).
	HPLC: (45% MeOH, 10 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 294 nm (pH 7), ϵ = 40000 (est.).
	ESI-MS (+): m/z calculated for C₁₉H₁₈BrN₅O₇P ([M + H]⁺): 538.01, found: 538.
	ESI-MS (−): m/z calculated for C₁₉H₁₆BrN₅O₇P ([M − H]⁻): 536.00, found: 536.

153

	β-(4-Aminophenyl)-1,N²-etheno-8-bromoguanosine-3′,5′-cyclic monophosphate
	(pNH₂-PET-8-Br-cGMP)
	The title compound was synthesized from 4-N₃-PET-8-Br-cGMP using general procedure V.
	Yield (Purity): 10% (>98%).
	HPLC: (18% MeCN, 50 mM NaH₂PO₄buffer, pH 6.7).
	UV-VIS: λ_max= 274 nm (pH 7), ϵ = 40000 (est.).
	ESI-MS (+): m/z calculated for C₁₈H₁₇BrN₆O₇P ([M + H]⁺): 539.01, found: 539.
	ESI-MS (−): m/z calculated for C₁₈H₁₇BrN₆O₇P ([M − H]⁻): 536.99, found: 537.

154

	8-Bromo-(6-methoxy-2-naphthyl-1,N²-etheno)guanosine-3′,5′-cyclic
	monophosphate (8-Br-(6-MeO-2-N)ET-cGMP)
	Using general procedure Y, 8-Br-cGMP was reacted with 2-bromo-6′-methoxy-2′-
	acethonaphtone to give the title compound.
	Yield (Purity): 6% (>99%).
	HPLC: (27% MeCN, 10 mM NaH₂PO₄buffer, pH 7.3).
	UV-VIS: λ_max= 260 nm (pH 7), ϵ = 40000 (est.).
	ESI-MS (+): m/z calculated for C₂₃H₂₀BrN₅O₈P ([M + H]⁺): 604.02, found: 604.
	ESI-MS (−): m/z calculated for C₁H₁N₅O₉PS ([M − H]⁻): 602.01, found: 602.

155

	8-Bromo-(9-phenanthrenyl-1,N²-etheno)guanosine-3′,5′-cyclic monophosphate
	(8-Br-(9-Phethr)ET-cGMP)
	Using general procedure Y, 8-Br-cGMP was reacted with 9-(2-bromoacetyl)phenantrene to
	give the title compound.
	Yield (Purity): 10% (>99%).
	HPLC: (28% MeCN, 50 mM NaH₂PO₄buffer, pH 6.7).
	HPLC (analytical): (26% MeCN, 50 mM NaH₂PO₄buffer, pH 7.2)
	UV-VIS: λ_max= 254 nm (pH 7), ϵ = 40000 (est.).
	ESI-MS (+): m/z calculated for C₂₆H₂₀BrN₅O₇P ([M + H]⁺): 624.03, found: 624.
	ESI-MS (−): m/z calculated for C₂₆H₁₈BrN₅O₇P ([M − H]⁻): 622.01, found: 622.

156

	8-Bromo-(4-trifluoromethyl-β-phenyl-1,N²-etheno)guanosine-3′,5′-cyclic
	monophosphate (8-Br-pTFMe-PET-cGMP)
	Using general procedure Y, 8-Br-cGMP was reacted with 2-bromo-4′-(trifluoromethyl)-
	acetophenon to give the title compound.
	Yield (Purity): 18% (>99%).
	HPLC: (27% MeCN, 50 mM NaH₂PO₄buffer, pH 6.7).
	UV-VIS: λ_max= 258 nm (pH 7), ϵ = 40000 (est.).
	ESI-MS (+): m/z calculated for C₁₉H₁₅BrF₃N₅O₇P ([M + H]⁺): 591.98, found: 592.
	ESI-MS (−): m/z calculated for C₁₉H₁₃BrF₃N₅O₇P ([M − H]⁻): 589.97, found: 590.

157

	(4-Fluoro-β-phenyl-1,N2-etheno)-8-methylthioguanosine-3′,5′-cyclic
	monophosphate (pF-PET-8-MeS-cGMP)
	Using general procedure A, 8-Br-pF-PET-cGMP (1 eq) was reacted with sodium
	methanethiolate (4 eq) to give the title compound.
	Yield (Purity): 30% (>99%).
	HPLC: (18% MeCN, 50 mM NaH₂PO₄buffer, pH 6.7).
	UV-VIS: λ_max= 270 nm (pH 7), ϵ = 40000 (est.).
	ESI-MS (+): m/z calculated for C₁₉H₁₈FN₅O₇PS ([M + H]⁺): 510.06, found: 510.
	ESI-MS (−): m/z calculated for C₁₉H₁₆FN₅O₇PS ([M − H]⁻): 508.05, found: 508.

158

	(4-Methoxy-β-phenyl-1,N2-etheno)-8-methylthioguanosine-3′,5′-cyclic
	monophosphate (pMeO-PET-MeS-cGMP)
	Using general procedure A 8-Br-pMeO-PET-cGMP (1 eq) was reacted with sodium
	methanethiolate (4 eq) to give the title compound.
	Yield (Purity): 88% (>99%).
	HPLC: (18% MeCN, 50 mM NaH₂PO₄buffer, pH 6.7).
	UV-VIS: λ_max= 275 nm (pH 7), ϵ = 40000 (est.).
	ESI-MS (+): m/z calculated for C₂₀H₂₁N₅O₈PS ([M + H]⁺): 522.08, found: 522.
	ESI-MS (−): m/z calculated for C₂₀H₁₉N₅O₈PS ([M − H]⁻): 520.07, found: 520.

159

	1,N²-Etheno-8-(2-phenylethyl)thioguanosine-3′,5′-cyclic monophosphate (ET-8-
	PhEtT-cGMP)
	Using general procedure A, 8-Br-Et-cGMP (B 177) was reacted with 2-phenyethanethiol to
	give the title compound.
	Yield (Purity): 18% (>99%).
	HPLC: (18% MeCN, 50 mM NaH₂PO₄buffer, pH 6.7).
	UV-VIS: λ_max= 288 nm (pH 7), ϵ = 19900 (est.).
	ESI-MS (+): m/z calculated for C₂₀H₂₁N₅O₇PS ([M + H]⁺): 506.09, found: 506.
	ESI-MS (−): m/z calculated for C₂₀H₁₉N₅O₇PS ([M − H]⁻): 504.07, found: 504.

160

	(4-Methoxy-β-phenyl-1,N2-etheno)-8-propylthioguanosine-3′,5′-cyclic
	monophosphate (pMeO-PET-8-PrT-cGMP)
	Using general procedure A, 8-Br-pMeO-PET-cGMP (1 eq) was reacted with
	1-Propanethiol (40 eq) to give the title compound.
	Yield (Purity): 40% (>99%).
	HPLC: (22% MeCN, 50 mM NaH₂PO₄buffer, pH 6.7).
	UV-VIS: λ_max= 276 nm (pH 7), ϵ = 40000 (est.).
	ESI-MS (+): m/z calculated for C₂₂H₂₄N₅O₈PS ([M + H]⁺): 550.12, found: 550.
	ESI-MS (−): m/z calculated for C₂₂H₂₄N₅O₈PS ([M − H]⁻): 548.10, found: 548.

161

	β-1,N²-Acetyl-8-bromoguanosine-3′,5′-cyclic monophosphate (β-1,N²-Ac-8-Br-
	cGMP)
	Using general procedure Y2, 8-Br-cGMP was reacted with methyl bromoacetate to give the
	title compound.
	Yield (Purity): 34% (>99%).
	HPLC: (7% MeCN, 30 mM TEAF buffer, pH 6.7).
	UV-VIS: λ_max= 266 nm (pH 7), ϵ = 15000 (est.).
	ESI-MS (+): m/z calculated for C₁₂H₁₂N₅O₈PBr ([M + H]⁺): 463.96, found: 464.
	ESI-MS (−): m/z calculated for C₁₂H₁₀N₅O₈PBr ([M − H]⁻): 461.95, found: 462.

162

	8-Bromo-β-1,N²-butyrylguanosine-3′,5′-cyclic monophosphate (8-Br-δ-1,N²-But-
	cGMP)
	Using general procedure Y3, 8-Br-1-CPr-cGMP was transformed into the title compound.
	Yield (Purity): 50% (>99%).
	HPLC: (26% MeOH, 50 mM TEAF buffer, pH 6.7).
	UV-VIS: λ_max= 265 nm (pH 7), ϵ = 16200 (est.).
	ESI-MS (+): m/z calculated for C₁₄H₁₆N₅O₈PBr ([M + H]⁺): 491.99, found: 492.
	ESI-MS (−): m/z calculated for C₁₄H₁₄N₅O₈PBr ([M − H]⁻): 489.98, found: 490.

163

	8-Bromo-1-(3-carboxypropyl)guanosine-3′,5′-cyclic monophosphate (8-Br-1-CPr-
	cGMP)
	Using general procedure Z, 8-Br-cGMP was reacted with ethyl 4-bromobutyrate to give the
	corresponding ethylester of the title compound. The crude product was transformed into the
	titel compound without prior chromatographic workup using general procedure F.
	Yield (Purity): 62%, 2 steps (>99%).
	HPLC: (22% MeOH, 200 mM TEAF buffer, pH 6.7).
	UV-VIS: λ_max= 266 nm (pH 7), ϵ = 16200 (est.).
	ESI-MS (+): m/z calculated for C₁₄H₁₈N₅O₉PBr ([M + H]⁺): 510.00, found: 510.
	ESI-MS (−): m/z calculated for C₁₄H₁₆N₅O₉PBr ([M − H]⁻): 507.99, found: 508.

164

	1-[Aminomethyl-(pentaethoxy)-propylamidopropyl]-8-bromoguanosine-3′,5′-
	cyclic monophosphate (1-AM-(EO)₅-PrAmdPr-8-Br-cGMP)
	Using adapted general procedure L, 8-Br-1-CPr-cGMP was reacted with NH₂CH₂-PEG₅-
	(CH₂)₃-NH₂(3 eq) to receive the title compound.
	Yield (Purity): 53% (>99%).
	HPLC: (36% MeOH, 100 mM TEAF buffer, pH 6.8).
	UV-VIS: λ_max= 267 nm (pH 7), ϵ = 16200 (est.).
	ESI-MS (+): m/z calculated for C₂₈H₄₈N₇O₁₃PBr ([M + H]⁺): 800.22, found: 800.
	ESI-MS (+): m/z calculated for C₂₈H₄₆N₇O₁₃PBr ([M − H]⁻): 798.20, found: 798.

165

	1-Benzyl-8-bromoguanosine-3′,5′-cyclic monophosphate
	(1-Bn-8-Br-cGMP)
	Using general procedure Z, 8-Br-cGMP was reacted with benzyl bromide to give the title
	compound.
	Yield (Purity): 70% (>99%).
	HPLC: (16% MeCN, 100 mM NaH₂PO₄buffer, pH 6.7).
	UV-VIS: λ_max265 nm (pH 7), ϵ = 16200 (est.).
	ESI-MS (+): m/z calculated for C₁₇H₁₈N₅O₇PBr ([M + H]⁺): 514.01, found: 514.
	ESI-MS (−): m/z calculated for C₁₇H₁₆N₅O₇PBr ([M − H]⁻): 512.00, found: 512.

166

	2′-O-(2-Azidoacetyl)-8-bromo-β-phenyl-1,N²-ethenoguanosine-3′,5′-cyclic
	monophosphate (2′-O-(2-N₃Ac)-8-Br-PET-cGMP)
	Using general procedure ZZ, 8-Br-PET-cGMP was transformed into the title compound.
	Yield (Purity): 60% (>99%).
	HPLC: (22% MeCN, 50 mM NaH₂PO₄buffer, pH 6.8).
	UV-VIS: λ_max= 258 nm (pH 7), ϵ = 40000 (est.).
	ESI-MS (+): m/z calculated for C₂₀H₁₇N₈O₈PBr ([M + H]⁺): 607.01, found: 607.
	ESI-MS (−): m/z calculated for C₂₀H₁₅N₈O₈PBr ([M − H]⁻): 604.99, found: 605.

2. Activation of PKG Isoforms by cGMP Derivatives
Experimental Part
In vitro activation experiments with PKG isozymes Iα, Iß and II were performed with the commercially available luminescence assay ADP-Glo™ Kinase Assay (Cat. #V9101) from Promega Corporation (Madison, Wis., USA) according to the manufacturer's instruction manual (The ADP-Glo™ Kinase Assay Technical Manual #TM313), standardized and conducted by BIAFFIN GmbH & Co KG (Kassel, Germany). Luminescence detection was accomplished with a LUMlstar Optima microplate luminometer from BMG LABTECH GmbH (Ortenberg, Germany). Bovine PKG type Iα was purified from bovine lung. Human PKGIß and PKGII were expressed in Sf9 cells and purified by affinity chromatography.²Concentrations of enzymes given below refer to the dimeric form. VASPtide (GL Biochem Ltd., Shanghai, China) was used as PKG-selective phosphorylation substrate peptide.²
Assay Conditions:
PKG Iα (0.2 nM), 20 mM Tris (pH 7.4), 10 mM Mg₂Cl₂, 1 mg/mL BSA, 0.15 mM β-mercaptoethanol, 2.5% DMSO, 130 μM VASPtide, 50 μM ATP, room temperature, 60 min.
PKG Iß (0.15 nM), 20 mM Tris (pH 7.4), 10 mM Mg₂Cl₂, 1 mg/mL BSA, 2.5% DMSO, 130 μM VASPtide, 50 μM ATP, room temperature, 60 min.
PKG II (0.5 nM), 20 mM Tris (pH 7.4), 10 mM Mg₂Cl₂, 1 mg/mL BSA, 5 mM β-mercaptoethanol, 2.5% DMSO, 130 μM VASPtide, 50 μM ATP, room temperature, 120 min.
Different concentrations (10 μM to 6 μM) of the compounds of the invention and cGMP as reference compound were incubated with the respective PKG isozyme. To increase assay sensitivity in case of PKG II, cGMP and compounds of the invention were preincubated at room temperature for 30 min. The activation values of the compounds are expressed as relative PKG activation compared to cGMP with cGMP set as 1 for each kinase isozyme. The K_a-values of cGMP for half-maximal kinase activation were 28 nM for Iα, 425 nM for Iß and 208 nM for II.
Results
FIGS. 3 to 5 show that all tested PLMs produce significantly higher relative PKG activation for at least 2 of the 3 PKG isozymes compared to the reference compound cGMP. Furthermore, it has to be noted, that the applied standard assay conditions only allowed to determine increased activation potencies of up to 140-fold for PKG Iα, 2832-fold for PKG Iß and 416-fold for PKG II, which is due to the employed enzyme concentration in the assays and the phenomenon that the isozymes were activity-titrated in some cases by the highly active compounds of the invention. The actual PKG activation potentials of these particular compounds of the invention appear to be significantly higher and are therefore expressed as ≥140-fold for PKG Iα, ≥2832-fold for PKG Iß and ≥416-fold for PKG II. A careful and more detailed analysis of the results is provided in the detailed description of the invention-section.
3. 661W Cell Line: Assessment of Cell Death Using the Ethidium Homodimer Assay
Background
To test the effect of PKG activators, the 661W cell line was used and increase in cell death after treatment was assessed. The 661W cell line is a photoreceptor precursor cell line, immortalized with the SV40 T antigen. As shown in the FIG. 6, the 661W cells express PKG. This makes them a suitable model for examining PKG activity using cell death as readout since increased PKG activity was previously associated with increased cell deaths. Because of potentially complex outcomes from the activation of different PKG isoforms this analysis is interpreted as a proof of principle on the use of these compounds in PKG-expressing cells or tissues.

Experimental Part

The 661W cells were cultured in DMEM with 10% FBS (Fetal Bovine Serum), 2 mM Glutamine, 100 U/ml penicillin, and 100 μg/ml streptomycin. For the Ethidium Homodimer Assay, cells were plated in a 24 well plate on slides coated with ECM (extracellular matrix) at 20 000 cells/well and left for 24 hours to attach to the slides. The next day the cells were treated with the compounds. Compounds were dissolved in water and then diluted in the medium at concentrations of 1 nM to 10 μM. 16 hours after treatment cells were washed with PBS and fixed in 4% paraformaldehyde. Afterwards slides were dipped into 2 μM Ethidium Homodimer for 2 minutes and nuclei were stained with DAPI (4′,6-diamidino-2-phenylindole). Ethidium Homodimer stains nuclei of dying cells. To assess cell death, microphotographs were taken from three different slides for each compound concentration and the total number of cells, as well as the number of dying Ethidium Homodimer positive cells, were counted in each picture. The value for untreated cells was set to 1. To statistically assess significant differences between untreated and treated cells, the unpaired Student's t-test was used and a P value ≤0.05 was considered significant (*≤0.05, **≤0.01, ***≤0.001).
Results
FIG. 7 shows percentage of cells undergoing cell death after treatment with non limiting exemplary polymer linked dimeric cGMP analogues of the invention (12 compounds). Six of the tested compounds led to significantly increased cell death at one or more concentrations when compared to untreated cells. The most potent compounds of the invention display a 5-6 fold increase in cell death when compared to untreated cells and 3-4 fold increase in cell death when compared to the reference 8-Br-PET-cGMP.


Acronyms

			spectrometry
		est.	estimated
BAP	bio-activatable protecting	Et₃NH⁺	triethylammonium
	group	FGF	fibroblast growth factor
cAMP	adenosine-3′,5′-cyclic	HCN	hyperpolarization-
	monophosphate		activated
			cyclic nucleotide-gated
cGMP	guanosine-3′,5′-cyclic	HEPES	4-(2-hydroxyethyl)-1-
	monophosphate		piperazineethanesulfonic
cGMPS	guanosine-3′,5′-cyclic		acid
	monophosphorothioate	HOBt	1-hydroxybenzotriazole
CNGC	cyclic nucleotide gated	HPLC	high performance liquid
	ion channel		chromatography
Cy	cyclohexyl	(i-Pr)₂EtNH⁺	diisopropylethy-
			lammonium
Cyp	cyclopentyl	i-PrOH	2-propanol
Da	Dalton	m/z	mass-to-charge ratio
DAPI	4′,6-diamidino-2-	MeCN	acetonitrile
	phenylindole	MTBE	tert-butyl methyl ether
DBU	1,8-	M_w	molecular weight
	diazabicyclo[5.4.0]	N2	N2-supplement for cell
	undec-7-ene		culture
DMEM/	Dulbecco's modifiziertes	NHS	N-hydroxysuccinimid
F12	eagle medium in	PAP	photo-activatable
			protecting
	combination with Ham′s		group
	F-12 medium	PBS	phosphate buffered
			saline
DMF	N,N-dimethylformamide	Pd(dppf)Cl₂	1,1′-
DMSO	dimethyl sulfoxide		bis(diphenylphosphino)
			ferro
dppf	1,1′-		cene-palladium(I1)
			dichloride
	Bis(diphenylphosphino)	PDE	phosphodiesterase
	ferro cene	PEG	poly(ethylene glycol)
ECM	extracellular matrix	PET	ß- Phenyl- 1, N²- etheno
EDC	1-ethyl-3-(3-	PFA	paraformaldehyde
	dimethylaminopropyl)	PKG	cGMP-dependent
	carbod iimide		protein kinase
EGTA	ethylene glycol-bis(2-	PLD	polymer linked dimer
	aminoethylether)-	PLM	polymer linked
	N,N,N′N′- tetraacetic		multimer
	acid	PN	postnatal
ESI-MS	electrospray Ionization	PyBOP	benzotriazole-1-yl-
	mass		oxytripyrrolid-
			inophosphonium
			hexafluorophosphate
Rp	as in Rp-cGMPS refers	VS	vinylsulfone
	to configuration of the
	chiral phosphorus,	ε	extinction coefficient
	wherein R/S	λ_max	wavelength at which
	follows the Cahn-Ingold-		absorbance is highest
	Prelog rules while
	“p” stands
	for phosphorus.
RP-18	reversed phase octadecyl
	modified material

SD standard deviation
TEA triethylammonium
TEAF triethylammonium formate
UV-VIS ultraviolet and visible (spectroscopy)

LITERATURE

1. (a) Schwede, F.; Maronde, E.; Genieser, H.; Jastorff, B., Cyclic nucleotide analogs as biochemical tools and prospective drugs. Pharmacol Ther 2000, 87 (2-3), 199-226; (b) Schmidt, H. H.; Hofmann, F.; Stasch, J., In cGMP: Generators, Effectors and Therapeuti Implications, Springer-Verlag Heidelberg: Berlin, 2009; pp 447-506; (c) Schlossmann, J.; Schinner, E., cGMP becomes a drug target. Naunyn Schmiedebergs Arch Pharmacol 2012, 385 (3), 243-52.
2. Poppe, H.; Rybalkin, S. D.; Rehmann, H.; Hinds, T. R.; Tang, X. B.; Christensen, A. E.; Schwede, F.; Genieser, H.; Bos, J. L.; Doskeland, S. O.; Beavo, J. A.; Butt, E., Cyclic nucleotide analogs as probes of signaling pathways. Nat. Methods 2008, 5, pp 277-278.
3. Schmidt, H. H.; Hofmann, F.; Stasch, J., In cGMP: Generators, Effectors and Therapeutic Implications, Springer-Verlag Heidelberg: Berlin, 2009; pp 409-421.
4. Herfindal, L.; Krakstad, C.; Myhren, L.; Hagland, H.; Kopperud, R.; Teigen, K.; Schwede, F.; Kleppe, R.; Doskland, S. O., Introduction of Aromatic Ring-Containing Substituents in Cyclic Nucleotides Is Associated with Inhibition of Toxin Uptake by the Hepatocyte Transporters OATP 1B1 and 1B3. PLoS ONE 2014, 9 (4), e94926.
5. Kramer, R. H.; Karpen, J. W., Spanning binding sites on allosteric proteins with polymer-linked ligand dimers. Nature 1998, 395, 710-713.
6. Sekhar, K. R.; Hatchett, R. J.; Shabb, J. B.; Wolfe, L.; Francis, S. H.; Wells, J. N.; Jastorff, B.; Butt, E.; Chakinala, M. M.; Corbin, J. D., Relaxation of Pig Arteries by New and Potent cGMP Analogs that Selectively Activate Type Iα, Compared with Iß, cGMP-Dependent Protein Kinase. Mol. Pharmacol 1992, pp 103-108.
7. Strassmaier, T.; Karpen, J., Novel N7- and N1-substituted cGMP Derivatives Are Potent Activators of Cyclic Nucleotide-Gated Channels. J. Med. Chem. 2007, 50, 4186-4194.
8. Kramer, R. H.; Karpen, J. W. Multimeric Tethered Ligands and Their Use in Receptor-Ligand Interaction. WO 99/25384, 1999.
9. Paquet-Durand, F.; Hauck, S. M.; van Veen, T.; Ueffing, M.; Ekstrom, P., PKG activity causes photoreceptor cell death in two retinitis pigmentosa models. J Neurochem 2009, 108 (3), 796-810.
10. (a) Wang, Y.; Chen, Y.; Wu, M.; Lan, T.; Wu, Y.; Li, Y.; Qian, H., Type II cyclic guanosine monophosphate-dependent protein kinase inhibits Rad activation in gastric cancer cells. Oncol Left 2015, 10 (1), 502-508; (b) Zhu, M.; Yao, X.; Wu, M.; Qian, H.; Wu, Y.; Chen, Y., Type II cGMP-dependent protein kinase directly inhibits HER2 activation of gastric cancer cells. Mol Med Rep 2016, 13 (2), 1909-13.
11. (a) Bala, I.; Hariharan, S.; Kumar, M. N., PLGA nanoparticles in drug delivery: the state of the art. Crit Rev Ther Drug Carrier Syst 2004, 21 (5), 387-422; (b) Basu, S. C.; Basu, M., Liposome Methods and Protocols. Humana Press: 2002; (c) Gregoriadis, G., Liposome Technology. Informa Healthcare: 2006; (d) Paquet-Durand, F.; Gaillard, P. J.; Maringo, V.; Ekström, P.; Genieser, H.-G.; Rentsch, A. Targeted liposomal delivery of cGMP analogues.
12. Genieser, H.-G.; Walter, U.; Butt, E. Derivatives of cyclic guanosine-3′,5′-monophosphorothioate. U.S. Pat. No. 5,625,056, Apr. 29, 1997.
13. Freudenberg, K.; Eichel, H.; Leutert, F., Synthesen von Abkömmlingen der Amino-säuren. Berichte der deutschen chemischen Gesellschaft (A and B Series) 1932, 65 (7), 1183-1191.
14. Genieser, H.-G., New boranophosphate Analogues of cyclic nucleotides. WO/2012/130829, 2012.

Claims

1. A compound having the formula (I) or (II)

wherein:

G units G¹and G²independently are compounds of formula (III) and G units G³and G⁴independently from G¹and G²and independently from each other are compounds of formula (III) or absent, wherein in case of formula (II) G⁴is always absent if G³is absent

and wherein in formula (III)

X, Y and Z are N

R₁, R₄, R₅, R₇and R₅can be equal or individual for each G unit (G¹, G², G³and G⁴),

while

R₁can independently be H, halogen, azido, cyano, acyl, aracyl, nitro, alkyl, aryl, aralkyl, OH, O-alkyl, O-aryl, O-aralkyl, O-acyl, O-aracyl, SH, S-alkyl, S-aryl, S-aralkyl, S-acyl, S-aracyl, S(O)-alkyl, S(O)-aryl, S(O)-aralkyl, S(O)-acyl, S(O)-aracyl, S(O)₂-alkyl, S(O)₂-aryl, S(O)₂-aralkyl, S(O)₂-acyl, S(O)₂-aracyl, SeH, Se-alkyl, Se-aryl, Se-aralkyl, NR9R10, SiR13R14R15 wherein R9, R10, R13, R14, R15 independently from each other can be H, alkyl, aryl, aralkyl;

R₂is absent;

R₃is OH;

R₄can independently be absent, H, amino, alkyl, aralkyl, nitro, N-oxide, or can form together with R₃, Y and the carbon bridging Y and R₃an imidazole ring which can be unsubstituted or substituted with alkyl, aryl or aralkyl, or can form together with Y and R₅and the carbon bridging Y and R₅an imidazole ring which can be unsubstituted or substituted with alkyl, aryl or aralkyl, or can form together with Y and R₅and the carbon bridging Y and R₅an imidazolinone as depicted (structure IV, V, n=1) or an homologous ring (n=2 to 8) which each can be unsubstituted or substituted (not depicted) with alkyl, aryl or aralkyl;

R₅can independently be H, halogen, NR30R31, NH-carbamoylR32R33, wherein R30, R31, R32, R33, independently from each other can be H, alkyl, aryl, aralkyl, or can form together with R₄, Y and the carbon bridging Y and R₅an imidazole ring which can be unsubstituted or substituted with alkyl, aryl or aralkyl, or can form together with R₄, Y and the carbon bridging Y and R₅an imidazolinone ring as depicted (structure IV, V, n=

1) or an homologous ring (n=2 to 8) which each can be unsubstituted or substituted (not depicted) with alkyl, aryl or aralkyl;

R₆is OH;

R₇is O-carbamoyl-alkyl, O-carbamoyl-aryl, O-carbamoyl-aralkyl, OH, O-alkyl, O-aryl, O-aralkyl, O-acyl, SH, S-alkyl, S-aryl, S-aralkyl, borano (BH₃), methylborano, dimethylborano, cyanoborano (BH₂CN)S-PAP, O-PAP, S-BAP, or O-BAP,

wherein PAP is a photo-activatable protecting group, optionally PAP=o-nitrobenzyl, 1-(o-nitrophenyl)-ethylidene, 4,5-dimethoxy-2-nitro-benzyl, 7-dimethylamino-coumarin-4-yl (DMACM-caged), 7-diethylamino-coumarin-4-yl (DEACM-caged) and 6,7-bis(carboxymethoxy)coumarin-4-yl)methyl (BCMCM-caged),

and wherein BAP is a bio-activatable protecting group, optionally BAP=methyl, acetoxymethyl, pivaloyloxymethyl, methoxymethyl, propionyloxymethyl, butyryloxymethyl, cyanoethyl, phenyl, benzyl, 4-acetoxybenzyl, 4-pivaloyloxybenzyl, 4-isobutyryloxybenzyl, 4-octanoyloxybenzyl, 4-benzoyloxybenzyl;

and

R₅is O-carbamoyl-alkyl, O-carbamoyl-aryl, O-carbamoyl-aralkyl, OH, O-alkyl, O-aryl, O-aralkyl, O-acyl, O-PAP or O-BAP,

wherein PAP is a photo-activatable protecting group with non limiting examples of, optionally, PAP=o-nitro-benzyl, 1-(o-nitrophenyl)-ethylidene, 4,5-dimethoxy-2-nitro-benzyl, 7-dimethylamino-coumarin-4-yl (DMACM-caged), 7-diethylamino-coumarin-4-yl (DEACM-caged) and 6,7-bis(carboxymethoxy)coumarin-4-yl)methyl (BCMCM-caged);

and wherein BAP is a bio-activatable protecting group with non limiting examples of, optionally, BAP=methyl, acetoxymethyl, pivaloyloxymethyl, methoxymethyl, propionyloxymethyl, butyryloxymethyl, cyanoethyl, phenyl, benzyl, 4-acetoxybenzyl, 4-pivaloyloxybenzyl, 4-isobutyryloxybenzyl, 4-octanoyloxybenzyl, 4-benzoyloxybenzyl;

and wherein

linking residues LR¹, LR², LR³and LR⁴independently can replace or covalently bind to any of the particular residues R₁, R₄and/or R₅of the G units (G^1-4) they connect,

wherein in case the particular linking residue (LR^1-4) covalently binds to any of the residues R₁, R₄and/or R₅, an endstanding group of the particular residue (R₁, R₄and/or R₅), as defined above, is transformed or replaced within the assembled compound and is then further defined as part of the particular linking residue (LR^1-4) within the assembled compound,

while

LR¹is (a) a tri- or tetravalent branched hydrocarbon moiety or (b) a divalent hydrocarbon moiety each with or without incorporated heteroatoms, optionally heteroatoms O, N, S, Si, Se, B, wherein the backbone preferably contains 1 to 28 carbon atoms and can be, saturated or unsaturated, substituted or unsubstituted,

while

each attachment point independently can be a substituted or unsubstituted carbon- or heteroatom

and

in case poly ethylene glycole (PEG) moieties are incorporated in accordance to the definition, the preferred number of carbon atoms can be exceeded by the number present in the PEG moieties, wherein all PEG moieties together can contain a total amount of

1 to 500 ethylene glycol groups (—(CH₂CH₂O)_n— with n=1 to 500) in case of divalent linking residue (LR¹)

or

1 to 750 ethylene glycol groups (—(CH₂CH₂O)_n— with n=1 to 750) in case of trivalent linking residue (LR¹)

or

1 to 1000 ethylene glycol groups (—(CH₂CH₂O)_n— with n=1 to 1000) in case of tetravalent linking residue (LR¹),

and if substituted,

substituents include optionally one or more alkyl groups, halogen atoms, haloalkyl groups, (un)substituted aryl groups, (un)substituted heteroaryl groups, amino, oxo, nitro, cyano, azido, hydroxy, mercapto, keto, carboxy, carbamoyl, expoxy, methoxy, ethynyl,

and/or substituents can further be connected to each other, forming a ringsystem with 1 to 4 rings, with or without incorporated heteroatoms, saturated or unsaturated, substituted or unsubstituted, aliphatic or aromatic;

LR², LR³and LR⁴are divalent hydrocarbon moieties with or without incorporated heteroatoms, optionally heteroatoms O, N, S, Si, Se, B, wherein the backbone preferably contains 1 to 28 carbon atoms and can be, saturated or unsaturated, substituted or unsubstituted,

while

and

in case poly ethylene glycole (PEG) moieties are incorporated in accordance to the definition, the preferred number of carbon atoms can be exceeded by the number present in the PEG moieties, wherein all PEG moieties together can contain a total amount of 1 to 500 ethylene glycol groups (—(CH₂CH₂O)_n— with n=1 to 500) and, if substituted,

substituents include optionally one or more alkyl groups, halogen atoms, haloalkyl groups, (un)substituted aryl groups, (un)substituted heteroaryl groups, amino, oxo, nitro, cyano, azido, hydroxy, mercapto, keto, carboxy, carbamoyl, expoxy, methoxy, ethynyl

wherein in case of formula (II) if G⁴is absent, LR⁴is absent, too, and

wherein in case of formula (II) if G³and G⁴are absent, LR³and LR⁴are absent, too, and wherein

G¹, G², G³and G⁴can further be salts and/or hydrates

while optionally suitable salts of the particular phosphate moiety are lithium, sodium, potassium, calcium, magnesium, zinc or ammonium, and trialkylammonium, dialkylammonium, alkylammonium, e.g., triethylammonium, trimethylammonium, diethylammonium and octylammonium;

and wherein

G¹, G², G³and G⁴can optionally be isotopically or radioactively labeled, be PEGylated, immobilized or be labeled with a dye or another reporting group,

wherein

the reporting group(s) and/or dye(s)

(a) are coupled to G¹, G², G³and/or G⁴via a linking residue (LR⁵), bound covalently to or replacing any of the particular residues R₁, R₄and/or R₅independently for each G unit (G¹, G², G³and/or G⁴) while LR⁵can be as defined for LR²

or

(b) in case of formula (I) can replace G³and/or G⁴

and wherein

optionally suitable dyes include fluorescent dyes such as fluorescein, anthraniloyl, N-methylanthraniloyl, dansyl or the nitro-benzofurazanyl (NBD) system, rhodamine-based dyes such as Texas Red or TAMRA, cyanine dyes such as Cy™3, Cy™5, Cy™7, EVOblue™10, EVOblue™30, EVOblue™90, EVOblue™100 (EVOblue™-family), the BODIPY™-family, Alexa Fluor™-family, DY-547P1, DY-647P1, coumarines, acridines, oxazones, phenalenones, fluorescent proteins such as GFP, BFP and YFP, and near and far infrared dyes

and wherein

reporting groups optionally include quantum dots, biotin and tyrosylmethyl ester

and wherein

PEGylated refers to the attachment of a single or multiple LR^PEGgroup(s) independently, wherein LR^PEGcan be as defined for LR², with the provisos that in this case (i) of LR²only one terminus is connected to a G unit (G¹, G², G³and/or G⁴) by covalently binding to or replacing any of the particular residues R₁, R₄and/or R₅independently for each G unit (G¹, G², G³and/or G⁴), and (ii) the other terminus of LR²is either an alkyl group or a reactive group that allows for conjugation reactions and/or hydrogen bonding

while, optionally, non limiting examples of reactive groups are, —NH₂, —SH, —OH, —COOH, —N₃, —NHS-ester, halogen group, epoxide, ethynyl, allyl

and with the proviso (iii) that LR^PEGhas incorporated ethylene glycol moieties ((CH₂CH₂O)_n— with n=2 to 500)

with the proviso that the compound of formula (I) and/or (II), is not selected from

Guanosine-3′, 5′-cyclic monophosphate-[8-thioethylsulfonyl-(ethyloxy)_n-ethylsulfonylethylthio-8]-guanosine-3′, 5′-cyclic monophosphate (polydispers compound arising from synthesis with polydispers bis vinylsulfonyl-PEG_nwith M_wof 800 g/mol, 1.2 kg/mol, 3.4 kg/mol or 20 kg/mol; or monodispers compound with n=6).

2. A compound according to claim 1, wherein at least two G units are unequally substituted.

3. A compound according to claim 1, wherein

(a) in case of formula (I) G³and G⁴are absent

or

in case of formula (II) G³, G⁴, LR³and LR⁴are absent;

and wherein

R₄is not H and/or R₅is not NH₂

or

(b) in case of formula (I) G³and G⁴are absent

or

in case of formula (II) G³, G⁴, LR³and LR⁴are absent.

4. A compound according to claim 1 wherein

all R₇are SH and all R₅are O

or

all R₇are O and all R₅are OH.

5. A compound according to claim 1 wherein the linking residues LR¹, LR², LR³and LR⁴are further subdivided as depicted in formula (Ib) and (IIb),

wherein:

coupling functions C¹, C^1′, C², C^2′, C³, C^3′, C⁴and C^4′ independently from each other can be absent or as defined by structures selected from the group consisting of

while connectivity can be as depicted or reversed as exemplified by

G¹-O—C(O)—NH—S²versus G¹-NH—C(O)—O—S²

an wherein in case the coupling function (C¹, C^1′, C², C^2′, C³, C^3′, C⁴and/or C^4′) does not replace the residue of the G unit (R₁, R₄and/or R₅of G^1-4) but bind to it, the particular residue (R₁, R₄and/or R₅) involved in coupling of G units (or G unit with dye(s) or other reporting group(s)) independently from each other is

as defined in any of the preceding claims, wherein an endstanding group is replaced by or transformed to the coupling function

or

selected from the group depicted hereinafter (wherein if present, Q1 connects to the G unit)

and wherein

the linker (L) is selected from the group consisting of

Dimeric Linkers Trimeric Linkers Tetrameric linkers

while

n for each sidechain within a particular linker of the list herebefore can have an equal or individual value as defined

and

all chiral, diastereomeric, racemic, epimeric, and all geometric isomeric forms of linkers (L) of the list herebefore, though not explicitly depicted, are included herein

and

cationic linkers (L) such as ammonium-derivatives are salts containing chloride-, bromide-, iodide- phosphate-, carbonate-, sulfate-, acetate- or any other physiologically accepted counterion

and wherein

spacers (S¹, S², S³and S⁴) can be equal or individual within a particular compound, be absent or be —(CH₂)_n1—(CH₂CH₂ß)_m-(CH₂)_n2— (with ß=O, S or NH; m=1 to 500, n1=0 to 8, n2=0 to 8, while both n1 and n2 can independently be equal or individual) or —(CH₂)_n— (with n=1 to 24).

6. A compound according to claim 1

wherein

R₁can independently be H, halogen, azido, nitro, alkyl, acyl, aryl, OH, O-alkyl, O-aryl, SH, S-alkyl, S-aryl, S-aralkyl, S(O)-alkyl, S(O)-aryl, S(O)-aralkyl, S(O)-benzyl, S(O)₂-alkyl, S(O)₂-aryl, S(O)₂-aralkyl, amino, NH-alkyl, NH-aryl, NH-aralkyl, NR9R10, SiR13R14R15 wherein R9, R10, R13, R14, R15 are alkyl;

and/or

R₄can independently be absent, H, amino, alkyl, aralkyl, nitro, N-oxide or can form together with Y and R₅and the carbon bridging Y and R₅an imidazole ring which can be unsubstituted or substituted with alkyl, aryl or aralkyl, or can form together with Y and R₅and the carbon bridging Y and R₅an imidazolinone as depicted above (structure IV, V, n=1) or an homologous ring (n=2 to 8) which each can be unsubstituted or substituted (not depicted) with alkyl, aryl or aralkyl.

and/or

R₅can independently H, halogen, NH-carbamoyl-alkyl, NH-carbamoyl-aryl, NH-carbamoyl-aralkyl, amino, NH-alkyl, NH-aryl, NH-aralkyl, NR30R31 wherein R30 and R31 are alkyl, or can form together with R₄, Y and the carbon bridging Y and R₅an imidazole ring which can be unsubstituted or substituted with alkyl, aryl or aralkyl, or can form together with R₄, Y and the carbon bridging Y and R₅an imidazolinone ring as depicted above (structure IV, V, n=1) or an homologous ring (n=2 to 8) which each can be unsubstituted or substituted (not depicted) with alkyl, aryl or aralkyl.;

and/or

R₇is OH, O-alkyl, O-aryl, O-aralkyl, O-acyl, SH, S-alkyl, S-aryl, S-aralkyl, borano (BH₃), methylborano, dimethylborano, cyanoborano (BH₂CN),

and

R₈is OH, O-alkyl, O-aryl, O-aralkyl, O-acyl,

7. A compound according to claim 1

wherein

R₁is H, Cl, Br, I, F, N₃, NO₂, OH, SH, NH₂, CF₃, 2-furyl, 3-furyl, 2-bromo-5-furyl, (2-furyl)thio, (3-(2-methyl)furyl)thio, (3-furyl)thio, 2-thienyl, 3-thienyl, (5-(1-methyl)tetrazolyl)thio, 1,1,2-trifluoro-1-butenthio, (2-(4-phenyl)imidazolyl)thio, (2-benzothiazolyl)thio, (2,6-dichlorophenoxypropyl)thio, 2-(N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino)ethylthio, (4-bromo-2,3-dioxobutyl)thio, [2-[(fluoresceinylthioureido)amino]ethyl]thio, 2,3,5,6-tetrafluorophenylthio, (7-(4-methyl)coumarinyl)thio, (4-(7-methoxy)coumarinyl)thio, (2-naphtyl)thio, (2-(1-bromo)naphtyl)thio, benzimidazolyl-2-thiobenzothiazolylthio, 4-pyridyl, (4-pyridyl)thio, 2-pyridylthio, 5-amino-3-oxopentylamino, 8-amino-3,6-dioxaoctylamino, 19-amino-4,7,10,13,16-pentaoxanonadecylamino, 17-amino-9-aza-heptadecylamino, 4-(N-methylanthranoyl)aminobutylamino, dimethylamino, diethylamino, 4-morpholino, 1-piperidino, 1-piperazino, triphenyliminophosphoranyl or residue R₁is as depicted hereinafter as residue entry 1 or 2:

residue entry 1:

wherein

m=0-6;

Q=S, S(O), S(O)₂, O, NH, Se, CH₂, C(O);

X₁, X₂and X₃can be equal or independently be H, OH, NH₂, N₃, SH, CN, NO₂, F, Cl, Br, I, (CH₂)_nCH₃(with n=0-5), i-Pr, t-Bu, (CH₂)_nC≡CH (with n=0-5), (CH₂)_nC═CH₂(with n=0-5), CH₂OH, (CH₂)_nOCH₃(with n=1-2), CH₂N(CH₃)₂, O(CH₂)_nCH₃(with n=0-5), Oi-Pr, OCy, OCyp, OBn, OC(O)CH₃, OC(O)Ph, OCF₃, N(CH₃)₂, NH(CH₂)_nCH₃(with n=0-5), NHC(O)t-Bu, NHC(O)Ph, NHC(O)Ot-Bu, NHC(O)CH₃, NHC(O)CH₂N₃, B(OH)₂, CF₃, C(O)OH, C(O)OCH₃, C(O)Oi-Pr, C(O)Ot-Bu, C(O)OPh, C(O)OBn, C(O)NH₂, C(O)N(CH₃)₂, C(O)NHPh, C(O)NHBn, C(O)CF₃, CH₂C(O)OH, CH₂C(O)OCH₃, CH₂C(O)Oi-Pr, CH₂C(O)Ot-Bu, CH₂C(O)OBn, S(CH₂)_nCH₃(with n=0-5), S(CH₂)_nOEt (with n=1-4), SBn, SO₂CH₃, SO₂CF₃,

(with Y₁═H, SH, CN, Ph, F, CH₃, OCH₃, SCH₃, 4-thiophenyl, NO₂, pentyl),

(with Y₂═H, SH, F),

(with Y₃═H, SH),

residue entry 2:

wherein

m=0-6;

n=1-6;

Q=S, S(O), S(O)₂, O, NH, Se;

and/or

R₄is absent, amino, N-oxide or residue R₄is as depicted hereinafter as residue entry 1, 2, 3 or 4:

residue entry 1:

wherein

m=1-6;

X₁, X₂and X₃can be equal or independently be H, N₃, CN, NO₂, F, Cl, Br, I, (CH₂)_nCH₃(with n=0-5), i-Pr, t-Bu, (CH₂)_nC≡CH (with n=0-5), (CH₂)_nC═CH₂(with n=0-5), (CH₂)_nOCH₃(with n=1-2), CH₂N(CH₃)₂, O(CH₂)_nCH₃(with n=0-5), Oi-Pr, OBn, OC(O)CH₃, OC(O)Ph, OCF₃, N(CH₃)₂, NHC(O)t-Bu, NHC(O)Ph, NHC(O)Ot-Bu, NHC(O)CH₃, NHC(O)CH₂N₃, CF₃, C(O)OCH₃, C(O)Oi-Pr, C(O)Ot-Bu, C(O)OPh, C(O)OBn, C(O)NH₂, C(O)N(CH₃)₂, C(O)NHPh, C(O)NHBn, C(O)CF₃, CH₂C(O)OCH₃, CH₂C(O)Oi-Pr, CH₂C(O)Ot-Bu, CH₂C(O)OBn, S(CH₂)_nCH₃(with n=0-5), S(CH₂)_nOEt (with n=1-4), SBn, SPh, SO₂CF₃,

(with Y₁═H, CN, Ph, F, CH₃, OCH₃, SCH₃, NO₂, pentyl),

(with Y₂═H, F),

residue entry 2:

wherein

X₁can be H, CH₃, Ph;

X₂can be H, Ph, 2-naphtyl, 9-phenanthryl, 1-pyrenyl, 2,3-dihydro-1,4-benzodioxin-6-yl, dibenzo[b,d]furan-2-yl, 2,3-dihydro-1-benzofuran-5-yl, 1-benzothien-5-yl, 1-benzofuran-5-yl, cyclopropyl, 1-adamantyl, C(Ph)₃, 2-thienyl, 3-chloro-2-thienyl, 3-thienyl, 1,3-thiazol-2-yl, 2-pyridinyl, 5-chloro-2-thienyl, 1-benzofuran-2-yl,

X₃, X₄and X₅can independently be H, OH, NH, CH₃, Cl, Br, F, CN, N₃, CF₃, OCF₃, NO₂, C(O)OH, C(O)OCH₃, OCH₃, OBn, O-benzoyl, SCH₃, t-Bu, N(CH₃)₂, S-phenyl, Ph, S(O)₂CH₃, C(O)NH₂, NHS(O)₂CH₃;

residue entry 3:

wherein deviating from the definition above, any hydrogen atom attached to any of the ring carbon atoms including depicted, implied, or expressly defined hydrogen, or both hydrogen atoms (m=2) attached to the same particular carbon atom, can be replaced by one or two (equal) “floating groups” X₁respectively, as long as a stable structure is formed,

while m=1 or 2;

n=1-4;

X₁can be H, CH₃, Et, Pr, i-Pr, Bu, F, Ph, (CH₂)₂OH*,

Only for first case;

residue entry 4:

wherein

m=1-6;

n=1-6.

and/or

R₅is H, NH₂, F, Cl, Br, I, methylamino, NH-benzyl, NH-phenyl, NH-4-azidophenyl, NH-phenylethyl, NH-phenylpropyl, 2-aminoethylamino, n-hexylamino, 6-amino-n-hexylamino, 8-amino-3,6-dioxaoctylamino, dimethylamino, 1-piperidino, 1-piperazino or can form together with R₄, Y and the carbon bridging Y and R₅a ring system as depicted in the list hereinabove (entry 2 and 3);

and/or

R₇OH, methyloxy, ethyloxy, cyanoethyloxy, acetoxymethyloxy, pivaloyloxymethyloxy, methoxymethyloxy, propionyloxymethyloxy, butyryloxymethyloxy, acetoxyethyloxy, acetoxybutyloxy, acetoxyisobutyloxy, phenyloxy, benzyloxy, 4-acetoxybenzyloxy, 4-pivaloyloxybenzyloxy, 4-isobutyryloxybenzyloxy, 4-octanoyloxybenzyloxy, 4-benzoyloxybenzyloxy, acetyloxy, propionyloxy, benzoyloxy, SH, methylthio, acetoxymethylthio, pivaloyloxymethylthio, methoxymethylthio, propionyloxymethylthio, butyryloxymethylthio, cyanoethylthio, phenylthio, benzylthio, 4-acetoxybenzylthio, 4-pivaloyloxybenzylthio, 4-isobutyryloxybenzylthio, 4-octanoyloxybenzylthio, 4-benzoyloxybenzylthio, borano (BH₃), methylborano, dimethylborano, cyanoborano (BH₂CN);

and

R8 is OH, methyloxy, ethyloxy, cyanoethyloxy, acetoxymethyloxy, pivaloyloxymethyloxy, methoxymethyloxy, propionyloxymethyloxy, butyryloxymethyloxy, acetoxyethyloxy, acetoxybutyloxy, acetoxyisobutyloxy, phenyloxy, benzyloxy, 4-acetoxybenzyloxy, 4-pivaloyloxybenzyloxy, 4-isobutyryloxybenzyloxy, 4-octanoyloxybenzyloxy, 4-benzoyloxybenzyloxy, acetyloxy, propionyloxy, benzoyloxy;

8. A compound according to claim 1

wherein

R₁is H, Cl, Br, I, F, N₃, NO₂, OH, SH, NH₂, CF₃, 2-furyl, 3-furyl, (2-furyl)thio, (3-(2-methyl)furyl)thio, (3-furyl)thio, 2-thienyl, 3-thienyl, (5-(1-methyl)tetrazolyl)thio, 1,1,2-trifluoro-1-butenthio, (2-(4-phenyl)imidazolyl)thio, (2-benzothiazolyl)thio, (2,6-dichlorophenoxypropyl)thio, 2-(N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino)ethylthio, (4-bromo-2,3-dioxobutyl)thio, [2-[(fluoresceinylthioureido)amino]ethyl]thio, 2,3,5,6-tetrafluorophenylthio, (7-(4-methyl)coumarinyl)thio, (4-(7-methoxy)coumarinyl)thio, (2-naphtyl)thio, (2-(1-bromo)naphtyl)thio, benzimidazolyl-2-thio, benzothiazolylthio, 4-pyridyl, (4-pyridyl)thio, 2-pyridylthio, 5-amino-3-oxopentylamino, 8-amino-3,6-dioxaoctylamino, 19-amino-4,7,10,13,16-pentaoxanonadecylamino, 17-amino-9-aza-heptadecylamino, 4-(N-methylanthranoyl)aminobutylamino, dimethylamino, diethylamino, 4-morpholino, 1-piperidino, 1-piperazino, triphenyliminophosphoranyl or residue R₁is as depicted hereinafter as residue entry 1 or 2:

residue entry 1:

wherein

m=0-6;

Q=S, S(O), S(O)₂, NH;

X₁, X₂and X₃can be equal or independently be H, OH, NH₂, N₃, SH, CN, NO₂, F, Cl, Br, I, (CH₂)_nCH₃(with n=0-5), i-Pr, t-Bu, Ph, (CH₂)_nC≡CH (with n=0-5), (CH₂)_nC═CH₂(with n=0-5), CH₂OH, (CH₂)_nOCH₃(with n=1-2), CH₂N(CH₃)₂, O(CH₂)_nCH₃(with n=0-5), Oi-Pr, OCy, OCyp, OPh, OBn, OC(O)CH₃, OC(O)Ph, OCF₃, N(CH₃)₂, NH(CH₂)_nCH₃(with n=0-5), NHC(O)t-Bu, NHC(O)Ph, NHC(O)Ot-Bu, NHC(O)CH₃, NHC(O)CH₂N₃, B(OH)₂, CF₃, C(O)OH, C(O)OCH₃, C(O)Oi-Pr, C(O)Ot-Bu, C(O)OPh, C(O)OBn, C(O)NH₂, C(O)N(CH₃)₂, C(O)NHPh, C(O)NHBn, C(O)CF₃, CH₂C(O)OH, CH₂C(O)OCH₃, CH₂C(O)Oi-Pr, CH₂C(O)Ot-Bu, CH₂C(O)OBn, S(CH₂)_nCH₃(with n=0-5), S(CH₂)_nOEt (with n=1-4), SBn, SPh,

residue entry 2:

wherein

m=0-6;

n=1-6;

Q=S, S(O), S(O)₂, NH;

and/or

residue entry 1:

wherein

m=1-3;

X₁, X₂and X₃can be equal or independently be H, N₃, CN, NO₂, F, Cl, Br, I, (CH₂)_nCH₃(with n=0-5), i-Pr, t-Bu, Ph, (CH₂)_nC≡CH (with n=0-5), (CH₂)_nC═CH₂(with n=0-5), (CH₂)_nOCH₃(with n=1-2), CH₂N(CH₃)₂, O(CH₂)_nCH₃(with n=0-5), Oi-Pr, OPh, OBn, OC(O)CH₃, OC(O)Ph, OCF₃, N(CH₃)₂, NHC(O)t-Bu, NHC(O)Ph, NHC(O)Ot-Bu, NHC(O)CH₃, NHC(O)CH₂N₃, CF₃, C(O)OCH₃, C(O)Oi-Pr, C(O)Ot-Bu, C(O)OPh, C(O)OBn, C(O)NH₂, C(O)N(CH₃)₂, C(O)NHPh, C(O)NHBn, C(O)CF₃, CH₂C(O)OCH₃, CH₂C(O)Oi-Pr, CH₂C(O)Ot-Bu, CH₂C(O)OBn, S(CH₂)_nCH₃(with n=0-5), S(CH₂)_nOEt (with n=1-4), SBn, SPh, SO₂CF₃,

residue entry 2:

wherein

X₁can be H, CH₃, Ph;

X₂can be H, Ph, 2-naphtyl, 9-phenanthryl, 1-pyrenyl, 2,3-dihydro-1,4-benzodioxin-6-yl, dibenzo[b,d]furan-2-yl, 2,3-dihydro-1-benzofuran-5-yl, 1-benzothien-5-yl, 1-benzofuran-5-yl, cyclopropyl, 1-adamantyl, C(Ph)₃, 2-thienyl, 3-chloro-2-thienyl, 3-thienyl, 1,3-thiazol-2-yl, 2-pyridinyl, 1-benzofuran-2-yl;

residue entry 3:

wherein deviating from the definition above, any hydrogen atom attached to any of the ring carbon atoms including depicted, implied, or expressly defined hydrogen, or both hydrogen atoms (m=2) attached to the same particular carbon atom, can be replaced by one or two (equal) “floating groups” X₁respectively, as long as a stable structure is formed;

while m=1 or 2;

n=1-4;

X₁can be H, CH₃, Et, Pr, i-Pr, Bu, F, Ph, (CH₂)₂OH*,

* Only for first case;

residue entry 4:

or

wherein

m=1-6;

n=1-6;

and/or

residues involved in connecting a G unit with another G unit or a dye or another reporting group can be R1, R4 and/or R5 in which case the particular residue is

as defined just above (for the particular residue), wherein an endstanding group is transformed to or replaced by a coupling function

or

as depicted in the following list (if present Q1 connects to the G unit):

and/or

coupling functions (C^1-4and C^1′-4) are selected from the group consisting of

and/or

the linker (L) can be absent or as depicted in the following list of linker (L):

while n for each sidechain within a particular linker can have an equal or individual value as defined. and/or

R₇is OH, methyloxy, ethyloxy, cyanoethyloxy, acetoxymethyloxy, pivaloyloxymethyloxy, methoxymethyloxy, propionyloxymethyloxy, butyryloxymethyloxy, acetoxyethyloxy, acetoxybutyloxy, acetoxyisobutyloxy, phenyloxy, benzyloxy, 4-acetoxybenzyloxy, 4-pivaloyloxybenzyloxy, 4-isobutyryloxybenzyloxy, 4-octanoyloxybenzyloxy, 4-benzoyloxybenzyloxy, acetyloxy, propionyloxy, benzoyloxy, SH, methylthio, acetoxymethylthio, pivaloyloxymethylthio, methoxymethylthio, propionyloxymethylthio, butyryloxymethylthio, cyanoethylthio, phenylthio, benzylthio, 4-acetoxybenzylthio, 4-pivaloyloxybenzylthio, 4-isobutyryloxybenzylthio, 4-octanoyloxybenzylthio, 4-benzoyloxybenzylthio, borano (BH₃), methylborano, dimethylborano, cyanoborano (BH₂CN);

and

R₈is OH, methyloxy, ethyloxy, cyanoethyloxy, acetoxymethyloxy, pivaloyloxymethyloxy, methoxy, methyloxy, propionyloxymethyloxy, butyryloxymethyloxy, acetoxyethyloxy, acetoxybutyloxy, acetoxyisobutyloxy, phenyloxy, benzyloxy, 4-acetoxybenzyloxy, 4-pivaloyloxybenzyloxy, 4-isobutyryloxybenzyloxy, 4-octanoyloxybenzyloxy, 4-benzoyloxybenzyloxy, acetyloxy, propionyloxy, benzoyloxy;

9. A compound according to claim 1

wherein

R₁is H, Cl, Br, SH, 2-furyl, 3-furyl, (2-furyl)thio, (3-(2-methyl)furyl)thio, (3-furyl)thio, 2-thienyl, 3-thienyl, (5-(1-methyl)tetrazolyl)thio, 1,1,2-trifluoro-1-butenthio, (2-(4-phenyl)imidazolyl)thio, (2-benzothiazolyl)thio, (2,6-dichlorophenoxypropyl)thio, 2-(N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino)ethylthio, (4-bromo-2, 3-dioxobutyl)thio, [2-[(fluoresceinylthioureido)amino]ethyl]thio, 2,3,5,6-tetrafluorophenylthio, (7-(4-methyl)coumarinyl)thio, (4-(7-methoxy)coumarinyl)thio, (2-naphtyl)thio, (2-(1-bromo)naphtyl)thio, benzimidazolyl-2-thio, benzothiazolylthio, 4-pyridyl, (4-pyridyl)thio, 2-pyridylthio, triphenyliminophosphoranyl or residue R₁is as depicted hereinafter as residue entry 1 or 2:

residue entry 1:

wherein

m=0-3;

Q=S;

residue entry 2:

wherein

m=0-6;

n=1-6;

Q=S.

and/or

R₄is absent or as depicted hereinafter as residue entry 1, 2, 3 or 4:

residue entry 1:

wherein

m=1-3;

residue entry 2:

wherein

X₁can be H, CH₃, Ph;

residue entry 3:

wherein

n=1-4;

residue entry 4:

wherein

m=1-3;

n=1-6.

and/or

R₅is NH₂, or can form together with R₄, Y and the carbon bridging Y and R₅a ring system as depicted in the list hereinabove (entry 2 and 3);

and/or

residues involved in connecting a G unit with another G unit or a dye or another reporting group can be R₁, R₄and R₅, in which case the particular residue is

or

as depicted in the following list (if present Q1 connects to the G unit)

R₁ R₄ R₄+ R₅

and/or

coupling functions (C_1-4and C^1′-4′) are as depicted by structures selected from the group consisting of

and/or

R₇OH, methyloxy, ethyloxy, cyanoethyloxy, acetoxymethyloxy, pivaloyloxymethyloxy, methoxy, methyloxy, propionyloxymethyloxy, butyryloxymethyloxy, acetoxy ethyloxy, acetoxybutyloxy, acetoxyisobutyloxy, phenyloxy, benzyloxy, 4-acetoxybenzyl oxy, 4-pivaloyloxybenzyloxy, 4-isobutyryloxybenzyloxy, 4-octanoyloxybenzyloxy, 4-benzoyloxybenzyloxy, acetyloxy, propionyloxy, benzoyloxy, SH, methylthio, acetoxymethylthio, pivaloyloxymethylthio, methoxymethylthio, propionyloxymethylthio, butyryloxymethylthio, cyanoethylthio, phenylthio, benzylthio, 4-acetoxybenzylthio, 4-pivaloyloxybenzylthio, 4-isobutyryloxybenzylthio, 4-octanoyloxybenzylthio, 4-benzoyloxybenzylthio;

and

R₅is OH, methyloxy, ethyloxy, cyanoethyloxy, acetoxymethyloxy, pivaloyloxymethyloxy, methoxy, methyloxy, propionyloxymethyloxy, butyryloxymethyloxy, acetoxy ethyloxy, acetoxybutyloxy, acetoxyisobutyloxy, phenyloxy, benzyloxy, 4-acetoxybenzyloxy, 4-pivaloyloxybenzyloxy, 4-isobutyryloxybenzyloxy, 4-octanoyloxybenzyloxy, 4-benzoyloxybenzyloxy, acetyloxy, propionyloxy, benzoyloxy;

10. A compound according to claim 1 selected from the group consisting of

(1) Guanosine-3′, 5′-cyclic monophosphate-[8-thio-(pentaethoxy)-ethylthio-8]-guanosine-3′, 5′-cyclic monophosphate

(2) Guanosine-3′, 5′-cyclic monophosphate-[8-thiomethylamidomethyl-(pentaethoxy)propylamidomethylthio-8]-guanosine-3′, 5′-cyclic monophosphate

(3) Guanosine-3′, 5′-cyclic monophosphate-[8-thiomethylamido-(octaethoxy)ethylamidomethylthio-8]-guanosine-3′, 5′-cyclic monophosphate

(4) Guanosine-3′, 5′-cyclic monophosphate-[8-(4-thiophenylthio)-(pentaethoxy)-ethyl(4-thiophenylthio)-8]-guanosine-3′, 5′-cyclic monophosphate

(5) β-Phenyl-1, N²-ethenoguanosine-3′, 5′-cyclic monophosphate-[8-thiomethylamido(octaethoxy)-ethylamidomethylthio-8]-β-phenyl-1, N²-ethenoguanosine-3′, 5′-cyclic monophosphate

(6) 8-Bromoguanosine-3′, 5′-cyclic monophosphate-[1, N²-etheno-β-phenyl-4-yl-(1-[1, 2, 3]-triazole-4-yl)-methoxy-(hexaethoxy)-methyl-(4-(4-[1, 2, 3]-triazole-1-yl)-β-phenyl-1, N²-etheno)]-8-bromoguanosine-3′, 5′-cyclic monophosphate

(7) Guanosine-3′, 5′-cyclic monophosphate-[8-thiomethylamido-(octaethoxy)ethylamidomethylthio-8]-β-phenyl-1, N²-ethenoguanosine-3′, 5′-cyclic monophosphate

(8) Guanosine-3′, 5′-cyclic monophosphate-[8-thiomethylamido-(nonadecaethoxy)ethylamidomethylthio-8]-guanosine-3′, 5′-cyclic monophosphate

(9) Guanosine-3′, 5′-cyclic monophosphate-[8-(1-[1, 2, 3]-triazole-4-yl)-methoxy(hexaethoxy)-methyl-(4-[1, 2, 3]-triazole-1-yl)-8]-guanosine-3′, 5′-cyclic monophosphate

(10) Guanosine-3′, 5′-cyclic monophosphate-[8-thiomethylamido-(PEG pd 2000)amidomethylthio-8]-guanosine-3′, 5′-cyclic monophosphate

(11) β-Phenyl-1, N²-ethenoguanosine-3′, 5′-cyclic monophosphate-[8-thiomethylamido(nonadecaethoxy)-ethylamidomethylthio-8]-guanosine-3′, 5′-cyclic monophosphate

(12) β-Phenyl-1, N²-ethenoguanosine-3′, 5′-cyclic monophosphate-[8-thiomethylamido(nonadecaethoxy)-ethylamidomethylthio-8]-β-phenyl-1, N²-ethenoguanosine-3′, 5′-cyclic monophosphate

(13) β-Phenyl-1, N²-ethenoguanosine-3′, 5′-cyclic monophosphate-[8-thiomethylamido(PEG pd 2000)-amidomethylthio-8]-0-phenyl-1, N²-ethenoguanosine-3′, 5′-cyclic monophosphate

(14) Benzene-1, 3, 5-tri-[(8-amidomethyl-(pentaethoxy)propylamidomethylthio)guanosine-3′, 5′-cyclic monophosphate]

(15) Ethylene glycol-bis(2-aminoethylether)-N, N, N′, N′-tetra-[(8-methylamidoethylthio)guanosine-3′, 5′-cyclic monophosphate]

(16) Guanosine-3′, 5′-cyclic monophosphate-[8-thio-(dodecanyl)-thio-8]-guanosine-3′, 5′-cyclic monophosphate

(17) β-Phenyl-1, N²-ethenoguanosine-3′, 5′-cyclic monophosphate-[8-thio-(dodecanyl)thio-8]-0-phenyl-1, N²-ethenoguanosine-3′, 5′-cyclic monophosphate, triethyl ammonium salt

(18) Guanosine-3′, 5′-cyclic monophosphate-[8-thioethylamidomethyl-(1-[1, 2, 3]-triazole-4-yl)-methoxy-(hexaethoxy)-methyl-(4-[1, 2, 3]-triazole-1-yl)methylamidoethylthio-8]-guanosine-3′, 5′-cyclic monophosphate

(19) Guanosine-3′, 5′-cyclic monophosphate-[8-thioethylthio-8]-guanosine-3′, 5′-cyclic monophosphate

(20) Guanosine-3′, 5′-cyclic monophosphate-[8-thioethyl-(1-[1, 2, 3]-triazole-4-yl)methoxy-(hexaethoxy)-methyl-(4-[1, 2, 3]-triazole-1-yl)-ethylthio-8]-guanosine-3′, 5′-cyclic monophosphate

(21) Guanosine-3′, 5′-cyclic monophosphate-[8-thio-(dodecanyl)-(4-thiophenyl-4″-thiophenylthio)-(dodecanyl)-thio-8]-guanosine-3′, 5′-cyclic monophosphate

(22) Guanosine-3′, 5′-cyclic monophosphate-[8-thioethylamidomethyl-(1-[1, 2, 3]-triazole-4-yl)-methoxy-(hexaethoxy)-methyl-(4-(4-[1, 2, 3]-triazole-1-yl)-β-phenyl-1, N²-etheno)]-8-bromoguanosine-3′, 5′-cyclic monophosphate

(23) β-Phenyl-1, N²-ethenoguanosine-3′, 5′-cyclic monophosphate-[8-thioethyl-(1-[1, 2, 3]-triazole-4-yl)-methoxy-(hexaethoxy)-methyl-(4-[1, 2, 3]-triazole-1-yl)-ethylthio-8]-β-phenyl-1, N²-ethenoguanosine-3′, 5′-cyclic monophosphate

(24) 8-Bromoguanosine-3′, 5′-cyclic monophosphate-[1-propylamidomethyl(pentaethoxy)-propylamidomethylthio-8]-β-phenyl-1, N²-ethenoguanosine-3′, 5′-cyclic monophosphate

(25) 8-Bromoguanosine-3′, 5′-cyclic monophosphate-[1-(pentaethoxy)-ethyl-1]-8-bromoguanosine-3′, 5′-cyclic monophosphate

(26) 8-Bromoguanosine-3′, 5′-cyclic monophosphate-[1-propylamidomethyl(pentaethoxy)-propylamidopropyl-1]-8-bromoguanosine-3′, 5′-cyclic monophosphate

(27) 8-Bromoguanosine-3′, 5′-cyclic monophosphate-[1-propylamidomethyl(pentaethoxy)-propylamidomethylthio-8]-guanosine-3′, 5′-cyclic monophosphate

(28) Guanosine-3′, 5′-cyclic monophosphate-[8-(phenyl-4-thio)-(pentaethoxy)-ethyl-(4-thiophenyl)-8]-guanosine-3′, 5′-cyclic monophosphate

(29) β-1, N²—Acetyl-guanosine-3′, 5′-cyclic monophosphate-[8-thiomethylamido(octaethoxy)-ethylamidomethylthio-8]-β-1,N²-acetyl-guanosine-3′, 5′-cyclic monophosphate

(30) 8-Phenylguanosine-3′, 5′-cyclic monophosphate-[1, N²-etheno-0-phenyl-4-yl-(1-[1, 2, 3]-triazole-4-yl)-methoxy-(hexaethoxy)-methyl-(4-(4-[1, 2, 3]-triazole-1-yl)-β-phenyl-1, N²-etheno)]-8-phenylguanosine-3′, 5′-cyclic monophosphate

11. A monomeric compound of formula (III) and/or a monomeric precursor of formula (III) of a compound according to claim 1, wherein the monomeric compound of formula (III) and/or the monomeric precursor of formula (III) is defined as in any of the preceding claims, and preferably wherein the monomeric compound of formula (III) and/or the monomeric precursor of formula (III) complies with the following proviso:

R₇is O and R₈is OH

and further complies with at least one of the following provisos:

R₄is not H and R₅is NH₂

wherein R₄is attached via a —CH₂— bridge, which is part of R₄

or

R₅together with R₄, Y and the carbon bridging Y and R₅form a ring system, which can be

a) an imidazolinone ring as depicted hereinafter (n=1) or an homologous ring (n=2 to 8)

or

b) an imidazole ring, which can be unsubstituted or substituted as depicted hereinafter as residue entry 1 and 2

residue entry 1

wherein

X₁is H;

X₂can be H, 2-naphtyl, 9-phenanthryl, 1-pyrenyl, 2,3-dihydro-1,4-benzodioxin-6-yl, dibenzo[b,d]furan-2-yl, 2,3-dihydro-1-benzofuran-5-yl, 1-benzothien-5-yl, 1-benzofuran-5-yl, cyclopropyl, 1-adamantyl, C(Ph)₃, 2-thienyl, 3-chloro-2-thienyl, 3-thienyl, 1,3-thiazol-2-yl, 2-pyridinyl, 5-chloro-2-thienyl, 1-benzofuran-2-yl;

X₃, X₄and X₅can independently be OH, NH, CH₃, Cl, Br, F, CN, N₃, CF₃, OCF₃, NO₂, C(O)OH, C(O)OCH₃, OCH₃, OBn, O-benzoyl, SCH₃, t-Bu, N(CH₃)₂, S-phenyl, Ph, S(O)₂CH₃, C(O)NH₂, NHS(O)₂CH₃, while X₄and X₅can also independently be H;

residue entry 2

while R₁is as in any of the compounds 31 to 107.

or

R₁is attached via a —S(O)— or —S(O)₂— bridge or via a carbon atom of an aromatic ring system, which in each case is part of R₁

while R₄is H and R₅is NH₂

and in addition complies with the proviso that the monomeric compound of formula (III) and/or the monomeric precursor compound of formula (III) is not selected from the group of compounds consisting of

and/or

the monomeric compound of formula (III) and/or the monomeric precursor of formula (III) is selected from the group consisting of

(31) 8-Amidomethylthioguanosine-3′, 5′-cyclic monophosphate

(32) 8-(4-Boronatephenylthio)-guanosine-3′, 5′-cyclic monophosphate

(33) 8-(4-Cyanobenzylthio)guanosine-3′, 5′-cyclic monophosphate

(34) 8-(4-(2-Cyanophenyl)-benzylthio)guanosine-3′, 5′-cyclic monophosphate

(35) 8-Cyclohexylmethylthioguanosine-3′, 5′-cyclic monophosphate

(36) 8-(2, 4-Dichlorophenylthio)guanosine-3′, 5′-cyclic monophosphate

(37) 8-Diethylphosphonoethylthio-guanosine-3′, 5′-cyclic monophosphate

(38) 8-Ethylthioguanosine-3′, 5′-cyclic monophosphate

(39) 8-Hexylthioguanosine-3′, 5′-cyclic monophosphate

(40) 8-(4-Isopropylphenylthio)guanosine-3′, 5′-cyclic monophosphate

(41) 8-(3-(2-Methyl)furanyl)thioguanosine-3′, 5′-cyclic monophosphate

(42) 8-(5-(1-Methyl)tetrazolyl)thioguanosine-3′, 5′-cyclic monophosphate

(43) 8-(4-Methoxybenzylthio)guanosine-3′, 5′-cyclic monophosphate

(44) 8-(7-(4-Methyl)coumarinyl)thio-guanosine-3′, 5′-cyclic monophosphate

(45) 8-Methylacetylthioguanosine-3′, 5′-cyclic monophosphate

(46) 8-(5-(1-Phenyl)tetrazolyl)thioguanosine-3′, 5′-cyclic monophosphate

(47) 8-(2-Phenylethyl)thioguanosine-3′, 5′-cyclic monophosphate

(48) 8-(2-(4-Phenyl)imidazolyl)thioguanosine-3′, 5′-cyclic monophosphate

(49) 8-(2-Thiophenyl)thioguanosine-3′, 5′-cyclic monophosphate

(50) 8-(1, 1, 2-Trifluoro-1-butenthio)guanosine-3′, 5′-cyclic monophosphate

(51) 8-Amidopropylthioguanosine-3′, 5′-cyclic monophosphate

(52) 8-Amidoethylthioguanosine-3′, 5′-cyclic monophosphate

(53) 8-Amidobutylthioguanosine-3′, 5′-cyclic monophosphate

(54) 8-Acetamidoethylthioguanosine-3′, 5′-cyclic monophosphate

(55) 8-(2-Benzothiazolyl)thioguanosine-3′, 5′-cyclic monophosphate

(56) 8-(2-Boronatebenzylthio)guanosine-3′, 5′-cyclic monophosphate

(57) 8-(4-Boronatebutylthio)guanosine-3′, 5′-cyclic monophosphate

(58) 8-(4-Boronatebenzylthio)guanosine-3′, 5′-cyclic monophosphate

(59) 8-(3-Boronatebenzylthio)guanosine-3′, 5′-cyclic monophosphate

(60) 8-Azidomethylamidoethylthio-guanosine-3′, 5′-cyclic monophosphate

(61) 8-(3-Boronatephenyl)amidobutylthio-guanosine-3′, 5′-cyclic monophosphate

(62) 8-Benzylamidobutylthioguanosine-3′, 5′-cyclic monophosphate

(63) 8-Benzamidoethylthioguanosine-3′, 5′-cyclic monophosphate

(64) 8-(3-Boronatephenyl)amidomethyl-thioguanosine-3′, 5′-cyclic monophosphate

(65) 8-Benzylamidomethylthio-guanosine-3′, 5′-cyclic monophosphate

(66) 8-(3-Boronatephenyl)amidoethylthio-guanosine-3′, 5′-cyclic monophosphate

(67) 8-(3-Boronatephenyl)amidopropylthioguanosine-3′, 5′-cyclic monophosphate

(68) 8-Carboxypropylthioguanosine-3′, 5′-cyclic monophosphate

(69) 8-Carboxybutylthioguanosine-3′, 5′-cyclic monophosphate

(70) 8-(2, 6-Dichlorophenoxypropyl)thio-guanosine-3′, 5′-cyclic monophosphate

(71) 8-(4-Dimethylaminophenyl)amido-methylthioguanosine-3′, 5′-cyclic monophosphate

(72) 8-(4-Dimethylaminophenyl)amido-butylthioguanosine-3′, 5′-cyclic monophosphate

(73) 8-Ethylbutyrylthioguanosine-3′, 5′-cyclic monophosphate

(74) 8-Methylpropionylthioguanosine-3′, 5′-cyclic monophosphate

(75) 8-Methylvalerianylthioguanosine-3′, 5′-cyclic monophosphate

(76) 8-Methoxyethylamidobutylthio-guanosine-3′, 5′-cyclic monophosphate

(77) 8-Methoxyethylamidomethylthio-guanosine-3′, 5′-cyclic monophosphate

(78) 8-Methoxyethylamidoethylthio-guanosine-3′, 5′-cyclic monophosphate

(79) 8-Phenylamidomethylthio-guanosine-3′, 5′-cyclic monophosphate

(80) 8-Phenylpropylthioguanosine-3′, 5′-cyclic monophosphate

(81) 8-(3-Butynylthio)guanosine-3′, 5′-cyclic monophosphate

(82) 8-(4-Acetamidophenylthio)guanosine-3′, 5′-cyclic monophosphate

(83) 8-(4-Chlorophenylsulfonyl)guanosine-3′, 5′-cyclic monophosphate

(84) 8-(4-Chlorophenylsulfoxide)-guanosine-3′, 5′-cyclic monophosphate

(85) 8-((2-Ethoxyethyl)-4-thiophenylthio)guanosine-3′, 5′-cyclic monophosphate

(86) 8-(4-Thiophenyl-4″-thiophenylthio)guanosine-3′, 5′-cyclic monophosphate

(87) 8-(2-Azidoethylthio)guanosine-3′, 5′-cyclic monophosphate

(88) 8-(3-Aminopropyl)-(pentaethoxy)-methylamidomethylthio-guanosine-3′, 5′-cyclic monophosphate

(89) 8-(2-Aminoethyl)-(octaethoxy)-amidomethylthioguanosine-3′, 5′-cyclic monophosphate

(90) 8-(2-Bromoethyl)-(pentaethoxy)-(4-thiophenylthio)guanosine-3′, 5′-cyclic monophosphate

(91) 8-(4-(Propargyloxy-(hexaethoxy)-methyl)-[1, 2, 3]-triazole-1-yl)methylamidoethylthio guanosine-3′, 5′-cyclic monophosphate

(92) 8-(4-Carboxyphenylthio)guanosine-3′, 5′-cyclic monophosphate

(93) 8-(4-Hydroxyphenylsulfonyl)-guanosine-3′, 5′-cyclic monophosphate

(94) 8-(4-Isopropylphenylsulfonyl)-guanosine-3′, 5′-cyclic monophosphate

(95) 8-(4-Methylcarboxyphenylthio)-guanosine-3′, 5′-cyclic monophosphate

(96) 8-Methylsulfonylguanosine-3′, 5′-cyclic monophosphate

(97) 8-(1-Bromo-2-naphthyl)methylthioguanosine-3′, 5′-cyclic monophosphate

(98) 8-(2-(1-Benzyl-[1, 2, 3]-triazole-4-yl)-ethylthio)guanosine-3′, 5′-cyclic monophosphate

(99) 8-(3-Fluoro-5-methoxybenzylthio)guanosine-3′, 5′-cyclic monophosphate

(100) 8-Pentafluorobenzylthioguanosine-3′, 5′-cyclic monophosphate

(101) 8-Triphenyliminophosphoranyl-guanosine-3′, 5′-cyclic monophosphate

(102) 8-(4-Chlorophenyl)guanosine-3′, 5′-cyclic monophosphate

(103) 8-(4-Fluorophenyl)guanosine-3′, 5′-cyclic monophosphate

(104) 8-(2-Furyl)guanosine-3′, 5′-cyclic monophosphate

(105) 8-(4-Hydroxyphenyl)guanosine-3′, 5′-cyclic monophosphate

(106) 8-(4-Isopropylphenyl)guanosine-3′, 5′-cyclic monophosphate

(107) 8-Phenylguanosine-3′, 5′-cyclic monophosphate

(108) β-Phenyl-1, N²-etheno-8-thioguanosine-3′, 5′-cyclic monophosphate

(109) 8-(2-Aminophenylthio)-ß-phenyl-1, N²-ethenoguanosine-3′, 5′-cyclic monophosphate

(110) 8-Cyclohexylthioß-phenyl-1, N²-ethenoguanosine-3′, 5′-cyclic monophosphate

(111) 8-Cyclopentylthio-ß-phenyl-1, N²-ethenoguanosine-3′, 5′-cyclic monophosphate

(112) 8-(4-Methylphenylthio)-ß-phenyl-1, N²-ethenoguanosine-3′, 5′-cyclic monophosphate

(113) 8-(4-Methoxyphenylthio)-ß-phenyl-1, N²-ethenoguanosine-3′, 5′-cyclic monophosphate

(114) 8-(3-(2-Methyl)furanyl)thio-ß-phenyl-1, N²-ethenoguanosine-3′, 5′-cyclic monophosphate

(115) 8-(7-(4-Methyl)coumarinyl)thio-ß-phenyl-1, N²-ethenoguanosine-3′, 5′-cyclic monophosphate

(116) 8-(2-Naphthyl)thio-ß-phenyl-1, N²-ethenoguanosine-3′, 5′-cyclic monophosphate

(117) ß-Phenyl-1, N²-etheno-8-(2-thiophenyl)thioguanosine-3′, 5′-cyclic monophosphate

(118) ß-Phenyl-1, N²-etheno-8-(2-phenylethyl)thioguanosine-3′, 5′-cyclic monophosphate

(119) 8-Amidomethylthio-ß-phenyl-1, N²-ethenoguanosine-3′, 5′-cyclic monophosphate

(120) 8-Carboxymethylthio-ß-phenyl-1, N²-ethenoguanosine-3′, 5′-cyclic monophosphate

(121) 8-(4-Boronatephenylthio)-ß-phenyl-1, N²-ethenoguanosine-3′, 5′-cyclic monophosphate

(122) 8-Ethylthio-ß-phenyl-1, N2-ethenoguanosine-3′, 5′-cyclic monophosphate

(123) 8-(4-Fluorophenylthio)-ß-phenyl-1, N²-ethenoguanosine-3′, 5′-cyclic monophosphate

(124) 8-Methylthio-ß-phenyl-1, N²-ethenoguanosine-3′, 5′-cyclic monophosphate

(125) ß-Phenyl-1, N²-etheno-8-propylthio-guanosine-3′, 5′-cyclic monophosphate

(126) 8-Azidoethylthio-ß-phenyl-1, N²-ethenoguanosine-3′, 5′-cyclic monophosphate

(127) ß-Phenyl-1, N²-etheno-8-(4-trifluoromethylphenylthio)guanosine-3′, 5′-cyclic monophosphate

(128) 8-(4-Chlorophenylsulfonyl)-ß-phenyl-1, N²-ethenoguanosine-3′, 5′-cyclic monophosphate

(129) 8-(4-Isopropylphenylthio)-ß-phenyl-1, N²-ethenoguanosine-3′, 5′-cyclic monophosphate

(130) 8-(4-Isopropylphenylsulfonyl)-ß-phenyl-1, N²-ethenoguanosine-3′, 5′-cyclic monophosphate

(131) 8-(4-Chlorophenyl)-ß-phenyl-1, N²-ethenoguanosine-3′, 5′-cyclic monophosphate

(132) 8-(4-Hydroxyphenyl)-ß-phenyl-1, N²-ethenoguanosine-3′, 5′-cyclic monophosphate

(133) 8-(4-Isopropylphenyl)-ß-phenyl-1, N²-ethenoguanosine-3′, 5′-cyclic monophosphate

(134) 8-Bromo-(4-methoxy-ß-phenyl-1, N2-etheno)guanosine-3′, 5′-cyclic monophosphate

(135) 8-Bromo-(4-methyl-ß-phenyl-1, N2-etheno)guanosine-3′, 5′-cyclic monophosphate

(136) alpha-Benzoyl-beta-phenyl-1, N2-etheno-8-bromoguanosine-3′, 5′-cyclic monophosphate

(137) 8-Bromo-(4-chloro-ß-phenyl-1, N2-etheno)guanosine-3′, 5′-cyclic monophosphate (138) 8-Bromo-(3-nitro-ß-phenyl-1, N2-etheno)guanosine-3′, 5′-cyclic monophosphate

(139) 8-Bromo-(ß-tert.-butyl-1, N2-etheno)guanosine-3′, 5′-cyclic monophosphate (140) 8-Bromo-(2-methoxy-ß-phenyl-1, N2-etheno)guanosine-3′, 5′-cyclic monophosphate

(141) 8-Bromo-(3-methoxy-ß-phenyl-1, N2-etheno)guanosine-3′, 5′-cyclic monophosphate

(142) 8-Bromo-(2, 4-dimethoxy-ß-phenyl-1, N2-etheno)guanosine-3′, 5′-cyclic monophosphate

(143) 8-Bromo-(4-pyridinyl-1, N2-etheno)guanosine-3′, 5′-cyclic monophosphate

(144) 8-Bromo-(3-thiophen-yl-1, N2-etheno)guanosine-3′, 5′-cyclic monophosphate

(145) 8-Bromo-(4-fluoro-ß-phenyl-1, N2-etheno)guanosine-3′, 5′-cyclic monophosphate

(146) 8-Bromo-1, N2-ethenoguanosine-3′, 5′-cyclic monophosphate

(147) 8-Bromo-(3-hydroxy-ß-phenyl-1, N2-etheno)guanosine-3′, 5′-cyclic monophosphate

(148) 8-Bromo-(4-hydroxy-ß-phenyl-1, N²-etheno)guanosine-3′, 5′-cyclic monophosphate

(149) 8-Bromo-(ß-(2, 3-dihydro-1, 4-benzodioxin)-1, N2-etheno)guanosine-3′, 5′-cyclic monophosphate

(150) 8-Bromo-(4-methylsulfonamido-ß-phenyl-1, N2-etheno)guanosine-3′, 5′-cyclic monophosphate

(151) 8-Bromo-(4-cyano-β-phenyl-1, N²-etheno)guanosine-3′, 5′-cyclic monophosphate

(152) 8-Bromo-(α-phenyl-β-methyl-1, N²-etheno)guanosine-3′, 5′-cyclic monophosphate

(153) β-(4-Aminophenyl)-1, N²-etheno-8-bromoguanosine-3′, 5′-cyclic monophosphate

(154) 8-Bromo-(6-methoxy-2-naphthyl-1, N²-etheno)guanosine-3′, 5′-cyclic monophosphate

(155) 8-Bromo-(9-phenanthrenyl-1, N²-etheno)guanosine-3′, 5′-cyclic monophosphate

(156) 8-Bromo-(4-trifluoromethyl-β-phenyl-1, N²-etheno)guanosine-3′, 5′-cyclic monophosphate

(157) (4-Fluoro-ß-phenyl-1, N2-etheno)-8-methylthioguanosine-3′, 5′-cyclic monophosphate

(158) (4-Methoxy-ß-phenyl-1, N2-etheno)-8-methylthioguanosine-3′, 5′-cyclic monophosphate

(159) 1, N²-Etheno-8-(2-phenylethyl)thioguanosine-3′, 5′-cyclic monophosphate

(160) (4-Methoxy-ß-phenyl-1, N2-etheno)-8-propylthioguanosine-3′, 5′-cyclic monophosphate

(161) β-1, N²-Acetyl-8-bromoguanosine-3′, 5′-cyclic monophosphate

(162) 8-Bromo-δ-1, N²-butyrylguanosine-3′, 5′-cyclic monophosphate

(163) 8-Bromo-1-(3-carboxypropyl)guanosine-3′, 5′-cyclic monophosphate

(164) 1-[Aminomethyl-(pentaethoxy)-propylamidopropyl]-8-bromoguanosine-3′, 5′-cyclic monophosphate

(165) 1-Benzyl-8-bromoguanosine-3′, 5′-cyclic monophosphate

(166) 2′-O-(2-Azidoacetyl)-8-bromo-β-phenyl-1, N²-ethenoguanosine-3′, 5′-cyclic monophosphate

12. A compound according to claim 1 or a monomeric compound of formula (III) according to claim 11 for use in the treatment of a disease or disorder, preferably a disease or disorder selected from the group consisting of cancer, cardiovascular disease or disorder, or autoimmune disease or disorder, or neurodegenerative disease or disorder.

13. A compound according to claim 1 or a monomeric compound of formula (III) for use in the treatment of at least one of:

a) neurodegenerative diseases associated with insufficient synaptic function and learning and memory defects.

b) neuromuscular junction defects including motor neuron diseases (including Amyotrophic lateral sclerosis (ALS), Primary lateral sclerosis), also forms caused by certain infectious diseases (including paralytic Poliomyelitis)

c) cancer, including the initiation of cancer cell apoptosis and the prevention of metastasis

d) cardiovascular diseases, including hypertension, cardiac hypertrophy, angina pectoris, ischemia and stroke

e) parasitic diseases caused by trypanosomes, including Malaria, Chagas, sleeping sickness

f) borelliosis (lyme disease)

g) pulmonary diseases and conditions, such as pulmonary fibrosis and pulmonary hypertension

h) osteoporosis

i) Autoimmune diseases associated with an excessive proliferation of B- and T-cells, including multiple sclerosis, Crohn's disease, Hashimoto's disease, juvenile arthritis, myocarditis, and rheuma.

14. Method of compound according to claim 1 or of a monomeric compound of formula (III) as research tool compound, preferably as research tool compound in regard of a disease or disorder, preferably a disease or disorder selected from the group consisting of cancer, cardiovascular disease or disorder, or autoimmune disease or disorder, or neurodegenerative disease or disorder.