RU2815444C2 - Cxcl8-binding nucleic acids - Google Patents

Abstract

FIELD: biotechnology.

SUBSTANCE: present invention relates to biotechnology and can be used in medicine and diagnostics. Invention discloses an L-nucleic acid molecule containing a nucleotide sequence in a central fragment: 5’-GGAAGUACGUGGAAAGCCRA(Xu)RAGUGUGUCCCG-3’ [SEQ ID NO: 27], where Xu is U or is absent. L-nucleic acid according to the present invention is capable of specifically binding to human CXCL8 (IL-8), without interacting with other ELR-positive chemokines, and more preferably without binding with ELR-positive chemokines, which bind to receptors CXCR1 and/or CXCR2.

EFFECT: invention can be used as a diagnostic or drug for diagnosing or treating a disease characterized by the presence of CXCL8, which is directly or indirectly involved in the pathogenesis of the disease.

98 cl, 7 ex, 1 tbl, 9 dwg

Description

The present invention relates to a nucleic acid molecule capable of binding to CXCL8; to a nucleic acid molecule for its use in a method of treating and/or preventing a disease; to a nucleic acid molecule for use in a CXCL8 detection method; to a nucleic acid molecule to obtain a detection agent or biosensor; to a pharmaceutical composition containing a nucleic acid molecule; to the use of a nucleic acid molecule to obtain a medicinal product; to the use of a nucleic acid molecule to obtain a diagnostic agent, diagnostic agent or biosensor; to the use of a nucleic acid molecule for detecting CXCL8; to a set containing a nucleic acid molecule; to a method for detecting CXCL8 using a nucleic acid molecule; to a complex containing a nucleic acid molecule; to a method for screening an antagonist of CXCL8-mediated activity using a nucleic acid molecule; and a method for detecting a nucleic acid molecule.

CXCL8 (UniProtKB/Swiss-Prot PI 0145, IL8_HUMAN; SEQ ID NO: 2), also known as interleukin 8 (abbreviated IL-8), neutrophil-activating protein 1 (abbreviated NAP-1), monocyte neutrophil chemotaxis factor (abbreviated MDNCF ) or granulocyte chemotaxis protein (abbreviated GCP-1) is a small basic protein that belongs to the CXC subfamily of chemokines with pro-inflammatory and pro-angiogenic properties characterized by the presence of a glutamate-leucine-arginine (abbreviated ELR) motif at the N-terminus. ELR-positive chemokines CXCL1 (also known as growth-regulating protein alpha, Gro-alpha, protein with melanoma growth-promoting activity, MGSA, or NAP-3), CXCL2 (also known as Gro-beta or macrophage inflammatory protein 2- alpha, MIP2-alpha), CXCL3 (also known as Gro-gamma or MIP2-beta), CXCL5 (also known as epithelial neutrophil-activating protein 78, ENA-78 or weakly inducible cytokine B5), CXCL6 (also known as chemokine alpha- 3, CKA-3, granulocyte chemotaxis protein 2, GCP-2, or weakly inducible cytokine B6), CXCL7 (also known as platelet basic protein, PBP, leukocyte growth factor, LDGF, macrophage growth factor, MDGF, or weakly inducible cytokine B7) and CXCL8 are agonists of the CXCR2 receptor (also known as IL8RB, IL8R type 2, CD182, CDwl28b or GRO/MGSA receptor). CXCL6, CXCL7 and CXCL8 are agonists of the CXCR1 receptor (also known as IL8RA, IL8R type 1, CD181 or CDwl28a). CXCL8 binds to the CXCR1 and CXCR2 receptors with similar affinities at a concentration of approximately 4 nM. Binding of CXCL8 to CXCR1/2 triggers Gαi-dependent signaling pathways and, for example, induces neutrophil migration, degranulation, and oxidative burst. Receptor sensitivity can be regulated through phosphorylation, beta-arrestin recruitment, and receptor internalization (Ha, Theranostics 2017). CXCL7 has the highest homology to CXCL8 with 33 identical amino acids. Non-human primate (macaca mulata, canine ape) CXCL8 is 95% identical (73 amino acids out of 77 amino acids are identical) to human CXCL8. There is no CXCL8 orthologue in mice and rats.

CXCL8 is secreted by various cell types, including monocytes, macrophages, fibroblasts, endothelial and epithelial cells in response to, for example, pathogen-associated molecular pattern molecules (abbreviated PAMPs) (e.g. LPS), proinflammatory mediators (e.g. IL-1, IL-6 and TNF-α), hypoxia, reactive oxygen or environmental stressors (e.g. cigarette smoke) (Ha, Theranostics 2017). CXCL8 acts as a chemotactic and activating cytokine for neutrophils and monocytes and plays an important physiological role in host defense.

CXCL8 is used as a biomarker for diagnosis, assessment of disease status, prognosis, and therapeutic efficacy in infectious diseases with elevated CXCL8 levels observed in viral infections (eg, respiratory syncytial virus, herpes simplex virus, hepatitis B and C virus, human cytomegalovirus), bacterial infections (for example, Streptococcus Pneumoniae and Mycobacterium tuberculosis ) and fungal infections (for example, Candida albicans and Aspergillus fumigatus ). When children are infected with respiratory syncytial virus (abbreviated RSV), plasma CXCL8 levels correlate with disease severity. Thus, CXCL8 may serve as a biomarker for assessing the severity of RSV infection and designing treatment regimens. Combination with other immunological biomarkers, namely lymphocyte counts and plasma CCL5 levels, improves the accuracy of the assessment (Brand, Pediatr Res. 2013). Additional examples of infectious diseases in which CXCL8 can be used as a biomarker include neonatal sepsis (Zhou, PLoS One 2015), bacterial meningitis (Yao, Int J Clin Exp Med 2015), pneumonia (Morris, Thorax 2009) and neurosyphilis (Wang , Sci Rep 2016). CXCL8 can also be used as a biomarker in inflammatory diseases, including chronic prostatitis, acute pyelonephritis, cystic fibrosis and various autoimmune diseases (Shahzad, Int Arch Med 2010). In cancer patients, CXCL8 levels correlate with tumor burden and poor treatment outcome (Sanmamed, Clin Cancer Res. 2014). For example, CXCL8 correlates with lower survival in breast cancer (Fang, Anticancer Res. 2017), pancreatic cancer (Chen, World J. Gastroenterol 2012) and childhood sarcoma (Highfill, Sci Transl Med 2014). In melanoma, elevated baseline levels of CXCL8 correlate with refractoriness to combination anti-CTLA 4 antibody/chemotherapy treatment and poor patient outcome (Jamal, J Immunother Cancer 2017).

CXCL8 and its receptors are involved in the pathogenesis of several inflammatory diseases, including inflammatory diseases of the airways (eg, chronic obstructive pulmonary disease, acute respiratory distress syndrome, asthma, cystic fibrosis, pulmonary fibrosis), inflammatory skin diseases (eg, neutrophilic dermatoses, psoriasis, bullous pemphigoid), autoimmune diseases (eg, inflammatory bowel disease, ulcerative colitis, multiple sclerosis, rheumatoid arthritis), inflammatory nerve diseases (eg, Sweet's nervous disease, Alzheimer's disease), ischemic diseases (eg, stroke, myocardial infarction, ischemia and cerebral infarction brain) and other inflammatory diseases (for example, atherosclerosis) (Ha, Theranostics 2017; Russo, Expert Rev Clin Immunol 2014). The effectiveness of CXCL8 inhibition in the treatment of such inflammatory diseases is confirmed in animal models of the disease. For example, in rabbits, administration of anti-CXCL8 antibody eliminates LPS-induced dermatitis, LPS-induced pleurisy, LPS/IL-1-induced arthritis, IC-type glomerulonephritis, acute lung injury and reperfusion lung injury (Bao, Int Immunopharmacol 2010; Harada, J Leukoc Biol 1994). Blockade of CXCR1/2 reduces ischemic brain damage in a rat model (Garau, Cytokine 2005).

CXCL8 is involved in the development, progression, and treatment resistance of human cancers, including lung cancer, colon cancer, prostate cancer, pancreatic cancer, breast cancer, ovarian cancer, and melanoma, and promotes tumor growth and metastasis (Brat, neurooncol 2005; Ha, Theranotics 2017; Koch, Science 1992; Li, Angiogenesis 2005; Liu, Cytokine Growth Factor Rev 2016; Xu, Oncol Res 2000). Mechanisms of cancer stimulation by CXCL8 include stimulation of angiogenesis, self-renewal of cancer stem cells, conversion of epithelial cells to mesenchymal cells and formation of an immunosuppressive tumor microenvironment (Ginestier, J Clin Invest 2010; Visvader, Nat Rev Cancer 2008; David, Vaccines 2016). The effectiveness of CXCL8 inhibition in the treatment of human cancer is confirmed in animal models of the disease. For example, knockdown of CXCL8 reduces the growth of ovarian cancer cells that are resistant to chemotherapy (Merritt, J Natl Cancer Inst 2008). Inhibition of CXCL8 enhances the antiangiogenic responses produced by docetaxel in ovarian and prostate cancer models (Campbell, Pharmaceuticals 2013). Overexpression of CXCL8 in tumors deficient in phosphatase and tensin homolog (PTEN) levels results in increased cancer cell viability and contributes to the development of treatment-resistant tumors (Campbell, Pharmaceuticals 2013; Maxwell, Eur Urol 2013; Maxwell, Oncotarget 2014). Inhibition of CXCL8 signaling results in sensitization of PTEN-deficient as well as p53-mutant cancer cells in response to DNA-disrupting agent, antimetabolite, or androgen receptor targeting strategies (Campbell, Pharmaceuticals 2013). Blockade of CXCR2 increases the effectiveness of anti-PD-1 antibody as an immune checkpoint inhibitor in mouse models of rhabdomyosarcoma and pancreatic ductal adenocarcinoma (Highfill, Sci Transl Med 2014; Steele, Cancer Cell 2016).

Several compounds targeting CXCL8 or one or both of the corresponding receptors CXCR1 and CXCR2 are known in the art and have been successfully tested in in vivo models. Some of them have also been tested in clinical trials. CXCR1 and/or CXCR2 antagonists, namely reparixin, ladarixin, danirixin, navarixin, SX-682 and AZD-5069, have been tested in clinical trials for pancreatic islet transplantation, organ transplantation, type 1 diabetes, COPD, asthma, bronchiectasis, psoriasis, bullous pemphigoid and cancer. A CXCR1/2 antagonist, reparixin, has undergone additional clinical trials in patients with three-symptom negative metastatic breast cancer. The CXCR2 antagonist AZD-5069 was tested in combination with an anti-PD-11 antibody, namely durvalumab, or chemotherapy in solid cancers.

Although the CXCR1 and CXCR2 receptors share common properties, however, ELR-positive chemokines have different expression patterns, namely, in vivo distribution (extracellular matrix binding properties) and mechanisms of interaction with the receptors (Feniger-Barish, Cytokine 1999; Sawant, Sci Rep . 2016). The respective physiological functions of the various ELR-positive chemokine family members are still poorly understood. Thus, selective inhibition of disease-causing CXCL8 may potentially be more effective in treating the disease and reducing the risk of side effects compared with broad blockade of CXCR1/CXCR2. For example, CXCL8 is required for the maintenance of cancer stem cells, but not CXCL1 (Liotti F Et al. Stem Cells 2017; 35: 135-146). During infection, chemokines have overlapping activities. Therefore, inhibition of a wide range of ELR-positive chemokines (eg, simultaneous inhibition of CXCL8 and CXCL1) may be associated with an increased risk of infection (Yung SC and Murphy PM; frontiers in Immunology, September 2012, vol. 3, Art. 276). CXCR1/2 ligands share an N-terminal ELR motif that can serve as a binding epitope (described in US Pat. No. 9783605). The presence of this common structural feature in the CXCR1/2 ligands CXCL1, CXCL2, CXCL3, CXCL5, CXCL6, CXCL7 and CXCL8 presents particular challenges when attempting to identify a compound that will selectively bind to and inhibit CXCL8 without negatively affecting other ELRs -positive chemokines.

Antibodies have been identified that bind to CXCL8, but not to the ELR-positive chemokines CXCL1 (also known as growth regulatory protein alpha, GRO-alpha) and CXCL2 (also known as growth regulatory protein beta, GRO-beta) (Skov, J Immunol 2008). CXCL8-specific monoclonal antibodies have so far failed to provide any efficacy in clinical trials that may be associated with the pharmacological properties of this class of substance, including pharmacokinetic profile, biodistribution and elimination. For example, high levels of chemokine production may lead to rapid saturation of therapeutic antibodies and lack of efficacy using commonly used long dosing intervals. In addition, antibodies may not have a negative effect on the binding of chemokines to glycosaminoglycans, which would result in a lack of destruction of the chemokine gradient and insufficient inhibition of leukocyte migration and extravasation. Nucleic acids preferentially bind to glycosaminoglycan binding sites on proteins (Oberthur, Nat Commun 2015). Thus, other classes of nucleic acid-like substances may be even better for therapeutic targeting of CXCL8.

Nucleic acids are sensitive to degradation by enzymes (nucleases) present in physiological fluids. This lack of biostability makes it difficult to prepare nucleic acid-based drugs for human treatment, as well as the preparation of nucleic acid-based diagnostic agents for diagnosing and/or treating disease. Chemical modifications can improve the stability of nucleic acids, but they may not be sufficient to provide stability sufficient for drug use. For example, nucleic acid with 2'-fluoro-modified pyrimidines has a plasma half-life comparable to that of all DNA-based nucleic acids, and 2'-O-methyl modifications are required to achieve long half-lives (Kratschmer, Nucleic Acid Ther 2017). Macugen, the first aptamer approved for therapeutic use, was stabilized by a combination of 2'-fluoropyrimidines, 2'-O-methylpurines, and a 3'-cap. In addition, chemical modifications may increase the risk of side effects when used in drug formulation. Conversely, nucleic acids with the unnatural L configuration are highly stable and have been shown to be safe, well tolerated by patients, and effective in clinical trials (Ludwig, Leukemia 2017; Menne, Nephrol Dial Transplant 2016).

2'-Fluoropyrimidine RNA aptamers that bind to CXCL8 have been described previously (Sung, Biomaterials 2014). However, in functional assays, the high binding affinity of one of the described aptamers, namely aptamer 8A-35, did not lead to improved functional properties such as inhibition of CXCL8-induced neutrophil migration with an IC ₅₀ >125 nM. Semi-quantitative analysis showed no or low binding of the precursor aptamer to the ELR-positive chemokines CXCL1 (also known as GRO-alpha) and CXCL2 (also known as GRO-beta).

The problem underlying the present invention is to create a nucleic acid molecule, preferably an L-nucleic acid, that specifically interacts with CXCL8, and preferably does not specifically interact with other ELR-positive chemokines, and more preferably does not bind with ELR-positive chemokines. chemokines that bind to the CXCR1 and/or CXCR2 receptors.

Another problem underlying the present invention is to provide a nucleic acid molecule, preferably an L-nucleic acid molecule, that binds CXCL8 with high affinity and thereby effectively inhibits CXCL8-stimulated chemotaxis, preferably with an inhibition constant in the single digit range. nanomolar range, and more preferably in the picomolar range.

A further problem underlying the present invention is the production of a nucleic acid molecule, and preferably an L-nucleic acid molecule, for the purpose of preparing a medicament for the treatment of a human and/or animal disease, wherein such disease is characterized by the presence of CXCL8, which directly or indirectly involved in the pathogenesis of this disease, and preferably a drug that suppresses or inhibits CXCL8 to such an extent that it provides a therapeutic effect.

Another problem underlying the present invention is to obtain a nucleic acid molecule, and preferably an L-nucleic acid molecule, in order to obtain a diagnostic agent for identifying, diagnosing and/or treating a disease, wherein such disease is characterized by the presence of CXCL8, which directly or indirectly participates in the pathogenesis of this disease.

These and other problems underlying the present invention can be solved by using the subject matter of the invention as claimed in the accompanying independent claims. Preferred embodiments of the invention can be taken from the dependent claims.

More specifically, the problem underlying the present invention can be solved in its first aspect by using an L-nucleic acid molecule capable of binding to human CXCL8, wherein the L-nucleic acid molecule comprises a central nucleotide fragment, wherein the central nucleotide fragment comprises a nucleotide sequence

5'-GGAAGUACGUGGAAAGCCRA(X _U )RAGUGUGUCCCG-3' [SEQ ID. NO: 27], where X _U is U or is absent.

More specifically, the problem underlying the present invention can be solved in its second aspect by using an L-nucleic acid molecule of the first aspect, including any variant thereof, for use in a method of treating and/or preventing a disease.

More specifically, the problem underlying the present invention can be solved in its third aspect by using an L-nucleic acid molecule of the first aspect, including any variant thereof, for use in a CXCL8 detection method.

More specifically, the problem underlying the present invention can be solved in its fourth aspect by using an L-nucleic acid molecule of the first aspect, including any variant thereof, to produce a detection agent or biosensor.

More specifically, the problem underlying the present invention can be solved in its fifth aspect by using a pharmaceutical composition comprising an L-nucleic acid molecule according to the first aspect, including any variant thereof, and optionally an additional component, wherein the additional component is selected from the group, comprising a pharmaceutically acceptable excipient, a pharmaceutically acceptable carrier and a pharmaceutically active agent.

More specifically, the problem underlying the present invention can be solved in its sixth aspect by using the L-nucleic acid molecule according to the first aspect, including any variant thereof, to produce a drug.

More specifically, the problem underlying the present invention can be solved in its seventh aspect by using the L-nucleic acid molecule according to the first aspect, including any variant thereof, to produce a diagnostic agent, diagnostic agent or biosensor.

More specifically, the problem underlying the present invention can be solved in its eighth aspect by using the L-nucleic acid molecule according to the first aspect, including any variant thereof, for detecting CXCL8, and preferably human CXCL8.

More specifically, the problem underlying the present invention can be solved in its ninth aspect by using a CXCL8 detection kit, wherein the kit contains an L-nucleic acid molecule according to the first aspect, including any variant thereof and at least an instruction or reaction insert vessel.

More specifically, the problem underlying the present invention can be solved in its tenth aspect by using a method for detecting CXCL8 using an L-nucleic acid determined according to the first aspect, including any variant thereof, in a sample, wherein said method includes the steps of:

a) obtaining a sample with an unknown concentration of CXCL8;

b) contacting the sample or diluting it with a diagnostic agent, diagnostic agent or biosensor as defined in the seventh aspect, including any variant thereof;

c) measuring a signal using a diagnostic agent, diagnostic tool or biosensor as defined in the seventh aspect, including any variant thereof;

d) comparing, but not necessarily, the signal with a standard,

where optionally, the concentration of CXCL8 in a sample with an unknown concentration of CXCL8 is determined by comparison with signals obtained from at least one sample with a known concentration of CXCL8, and preferably at least one sample with a known concentration of CXCL8 is subjected to steps b)-c).

More specifically, the problem underlying the present invention can be solved in its eleventh aspect by using a complex comprising an L-nucleic acid molecule according to the first aspect, including any variant thereof and CXCL8, where preferably the complex is a crystalline complex.

More specifically, the problem underlying the present invention can be solved in its twelfth aspect by using a method for screening an antagonist of CXCL8-mediated activity, wherein said method includes the following steps:

- obtaining a candidate antagonist of CXCL8-mediated activity,

- producing an L-nucleic acid molecule according to the first aspect, including any variant thereof,

- creating a test system that will transmit a signal in the presence of an antagonist of CXCL8-mediated activity, and

- confirming that the candidate antagonist of CXCL8-mediated activity is an antagonist of CXCL8-mediated activity.

More specifically, the problem underlying the present invention can be solved in its 13th aspect by using a method for detecting an L-nucleic acid molecule according to the first aspect, including any variant thereof, in a sample, wherein said method includes the steps of:

a) providing a capture probe, wherein the capture probe is at least partially complementary to a first portion of the L-nucleic acid molecule of the first aspect, including any variant thereof, and a detection probe, wherein the detection probe is at least partially complementary to a second portion of the L molecule -nucleic acid according to the first aspect, including any variant thereof, or, alternatively, wherein the capture probe is at least partially complementary to the second part of the L-nucleic acid molecule according to the first aspect, including any variant thereof, and the detection probe is at least partially complementary the first part of the L-nucleic acid molecule according to the first aspect, including any variant thereof;

b) adding a capture probe and a detection probe, alone or in combination, to a sample containing an L-nucleic acid molecule according to the first aspect, including any variant thereof, or purportedly containing an L-nucleic acid molecule according to the first aspect, including any option;

c) causing the capture probe and the detection probe to interact, either simultaneously or sequentially in any order, with the L-nucleic acid molecule according to the first aspect, including any variant thereof;

d) optionally, determining whether the capture probe hybridizes to the L-nucleic acid molecule according to the first aspect, including any variant thereof, as described in step a); And

e) detecting the complex formed in step c) and consisting of an L-nucleic acid molecule according to the first aspect, including any variant thereof, a capture probe and a detection probe.

More specifically, the present invention can also be implemented in accordance with the following embodiments 1 to 103, where option 1 corresponds to the first aspect, option 47 corresponds to the second aspect, option 49 corresponds to the third aspect, option 50 corresponds to the fourth aspect, option 51 corresponds to the fifth aspect, option Option 53 corresponds to the sixth aspect, option 56 corresponds to the seventh aspect, option 79 corresponds to the eighth aspect, option 96 corresponds to the ninth aspect, option 97 corresponds to the tenth aspect, option 98 corresponds to the eleventh aspect, option 99 corresponds to the twelfth aspect, and option 100 corresponds to the 13th aspect:

Option 1 : An L-nucleic acid molecule capable of binding to human CXCL8, wherein the L-nucleic acid molecule contains a central nucleotide fragment, wherein the central nucleotide fragment contains the nucleotide sequence:

5'-GGAAGUACGUGGAAAGCCRA(X _U )RAGUGUGUCCCG-3' [SEQ ID. NO: 27], where X _U is U or is absent.

Option 2 : L-nucleic acid molecule according to option 1, where the central nucleotide fragment contains a nucleotide sequence selected from the group:

a) 5' GGAAGUACGUGGAAAGCCAAUGAGUGUGUCCCG 3' [SEQ. ID. NO: 28],

b) 5' GGAAGUACGUGGAAAGCCGAUGAGUGUGUCCCG 3' [SEQ. ID. NO: 29] and

c) 5' GGAAGUACGUGGAAAGCCGAAAGUGUGUCCCG 3' [SEQ. ID. NO: 30].

Option 3 : L-nucleic acid molecule according to option 2, where the central nucleotide fragment contains the nucleotide sequence:

5' GGAAGUACGUGGAAAGCCGAAAGUGUGUCCCG 3' [SEQ. ID. NO: 30] or 5' GGAAGUACGUGGAAAGCCAAUGAGUGUGUCCCG 3' [SEQ. ID. NO: 28], and preferably 5' GGAAGUACGUGGAAAGCCGAAAGUGUGUCCCG 3' [SEQ. ID. NO: 30].

Option 4 : An L-nucleic acid molecule according to any one of options 1 to 3, wherein the L-nucleic acid molecule contains, in the 5'→3' direction, a first terminal nucleotide fragment, a central nucleotide fragment and a second terminal nucleotide fragment, wherein:

the first terminal nucleotide fragment contains from three to six nucleotides and

the second terminal nucleotide fragment contains from three to six nucleotides,

where preferable:

the first terminal nucleotide fragment contains from five to six nucleotides and

the second terminal nucleotide fragment contains from five to six nucleotides,

where it is more preferable:

the first terminal nucleotide fragment contains five nucleotides, and

the second terminal nucleotide fragment contains five nucleotides.

Option 5 : L-nucleic acid molecule of option 4, where the first terminal nucleotide fragment contains the nucleotide sequence 5'-Z ₁ Z ₂ Z ₃ Z ₄ Z ₅ C-3', and the second terminal nucleotide fragment contains the nucleotide sequence 5'-GZ ₆ Z ₇ Z ₈ Z ₉ Z ₁₀ -3', where

Z ₁ is G or absent, Z ₂ is S or absent; Z ₃ is K or absent, Z ₄ is S; Z ₅ is D; Z ₆ represents H; Z ₇ represents S; Z ₈ is M or absent; Z ₉ is S or absent, and Z ₁₀ is C or absent;

where preferable:

a) Z ₁ represents G; Z ₂ represents S; Z ₃ represents K; Z ₄ represents S; Z ₅ represents D; Z ₆ represents H; Z ₇ represents S; Z ₈ represents M; Z ₉ represents S, and Z ₁₀ represents C; or

b) Z ₁ is missing; Z ₂ represents S; Z ₃ represents K; Z ₄ represents S; Z ₅ represents D; Z ₆ represents H; Z ₇ represents S; Z ₈ represents M; Z ₉ is S and Z ₁₀ is missing; or

c) Z ₁ is missing; Z ₂ is missing; Z ₃ represents K; Z ₄ represents S; Z ₅ represents D; Z ₆ represents H; Z ₇ represents S; Z ₈ represents M; Z ₉ is missing and Z ₁₀ is missing; or

d) Z ₁ is missing; Z ₂ is missing; Z ₃ is missing; Z ₄ represents S; Z ₅ represents D; Z ₆ represents H; Z ₇ represents S; Z ₈ missing; Z ₉ is missing and Z ₁₀ is missing; or

e) Z ₁ is missing; Z ₂ represents S; Z ₃ represents K; Z ₄ represents S; Z ₅ represents D; Z ₆ represents H; Z ₇ represents S; Z ₈ represents M; Z ₉ represents S and Z ₁₀ represents C; or

f) Z ₁ is missing; Z ₂ is missing; Z ₃ represents K; Z ₄ represents S; Z ₅ represents D; Z ₆ represents H; Z ₇ represents S; Z ₈ represents M; Z ₉ represents S, and Z ₁₀ represents C; or

g) Z ₁ is missing; Z ₂ is missing; Z ₃ is missing; Z ₄ represents S; Z ₅ represents D; Z ₆ represents H; Z ₇ represents S; Z ₈ represents M; Z ₉ represents S, and Z ₁₀ represents C; or

h) Z ₁ represents G; Z ₂ represents S; Z ₃ represents K; Z ₄ represents S; Z ₅ represents D; Z ₆ represents H; Z ₇ represents S; Z ₈ represents M; Z ₉ is S and Z ₁₀ is absent; or

i) Z ₁ represents G; Z ₂ represents S; Z ₃ represents K; Z ₄ represents S; Z ₅ represents D; Z ₆ represents H; Z ₇ represents S; Z ₈ represents M; Z ₉ is missing and Z ₁₀ is missing; or

j) Z ₁ represents G; Z ₂ represents S; Z ₃ represents K; Z ₄ represents S; Z ₅ represents D; Z ₆ represents H; Z ₇ represents S; Z ₈ missing; Z ₉ is missing and Z ₁₀ is missing; or

k) Z ₁ is missing; Z ₂ represents S; Z ₃ represents K; Z ₄ represents S; Z ₅ represents D; Z ₆ represents H; Z ₇ represents S; Z ₈ represents M; Z ₉ is missing and Z ₁₀ is missing; or

l) Z ₁ is missing; Z ₂ represents S; Z ₃ represents K; Z ₄ represents S; Z ₅ represents D; Z ₆ represents H; Z ₇ represents S; Z ₈ missing; Z ₉ is missing and Z ₁₀ is missing; or

m) Z ₁ is missing; Z ₂ is missing; Z ₃ represents K; Z ₄ represents S; Z ₅ represents D; Z ₆ represents H; Z ₇ represents S; Z ₈ represents M; Z ₉ is S and Z ₁₀ is absent; or

n) Z ₁ is missing; Z ₂ is missing; Z ₃ is missing; Z ₄ represents S; Z ₅ represents D; Z ₆ represents H; Z ₇ represents S; Z ₈ represents M; Z ₉ is S and Z ₁₀ is absent; or

o) Z ₁ is missing; Z ₂ is missing; Z ₃ represents K; Z ₄ represents S; Z ₅ represents D; Z ₆ represents H; Z ₇ represents S; Z ₈ missing; Z ₉ is missing and Z ₁₀ is missing; or

p) Z ₁ is missing; Z ₂ is missing; Z ₃ is missing; Z ₄ represents S; Z ₅ represents D; Z ₆ represents H; Z ₇ represents S; Z ₈ represents M; Z ₉ is missing and Z ₁₀ is missing.

Option 6 : L-nucleic acid molecule according to any of options 4-5, where:

a) the first terminal nucleotide fragment contains the nucleotide sequence 5'-CUGAC-3', and the second terminal nucleotide fragment contains the nucleotide sequence 5'-GUCAG-3';

b) the first terminal nucleotide fragment contains the nucleotide sequence 5'-GCUGAC-3', and the second terminal nucleotide fragment contains the nucleotide sequence 5'-GUCAGC-3'; or

c) the first terminal nucleotide fragment contains the nucleotide sequence 5'-GCUGAC-3', and the second terminal nucleotide fragment contains the nucleotide sequence 5'-GUGAGC-3';

d) the first terminal nucleotide fragment contains the nucleotide sequence 5'-GUGAC-3', and the second terminal nucleotide fragment contains the nucleotide sequence 5'-GUCAC-3';

e) the first terminal nucleotide fragment contains a nucleotide sequence of 5'-UGAC-3', and the second terminal nucleotide fragment contains a nucleotide sequence of 5'-GUCA-3';

f) the first terminal nucleotide fragment contains the nucleotide sequence 5'-GGAC-3', and the second terminal nucleotide fragment contains the nucleotide sequence 5'-GUCC-3';

g) the first terminal nucleotide fragment contains the nucleotide sequence 5'-GCAC-3', and the second terminal nucleotide fragment contains the nucleotide sequence 5'-GUGC-3';

h) the first terminal nucleotide fragment contains the nucleotide sequence 5'-GGUC-3', and the second terminal nucleotide fragment contains the nucleotide sequence 5'-GACC-3';

i) the first terminal nucleotide fragment contains the nucleotide sequence 5'-UGGC-3', and the second terminal nucleotide fragment contains the nucleotide sequence 5'-GCCA-3';

j) the first terminal nucleotide fragment contains the nucleotide sequence 5'-GAC-3', and the second terminal nucleotide fragment contains the nucleotide sequence 5'-GUC-3';

where preferable:

the first terminal nucleotide fragment contains the nucleotide sequence 5'-CUGAC-3', and the second terminal nucleotide fragment contains the nucleotide sequence 5'-GUCAG-3'; or

the first terminal nucleotide fragment contains the nucleotide sequence 5'-GCUGAC-3', and the second terminal nucleotide fragment contains the nucleotide sequence 5'-GUCAGC-3' or

the first terminal nucleotide fragment contains the nucleotide sequence 5'-GUGAC-3', and the second terminal nucleotide fragment contains the nucleotide sequence 5'-GUCAC-3';

where it is more preferable:

the first terminal nucleotide fragment contains the nucleotide sequence 5'-CUGAC-3', and the second terminal nucleotide fragment contains the nucleotide sequence 5'-GUCAG-3';

or

the first terminal nucleotide fragment contains the nucleotide sequence 5'-GUGAC-3', and the second terminal nucleotide fragment contains the nucleotide sequence 5'-GUCAC-3'.

Option 7 : The L-nucleic acid molecule according to any one of embodiments 1 to 6, wherein the L-nucleic acid molecule contains 40 to 45 nucleotides, preferably 42 to 45 nucleotides, and more preferably 42 nucleotides.

Option 8 : An L-nucleic acid molecule according to any one of embodiments 1 to 7, wherein the L-nucleic acid molecule is composed of ribonucleotides.

Option 9 : An L-nucleic acid molecule according to any one of embodiments 1-8, wherein the L-nucleic acid molecule contains a nucleotide sequence selected from the group of SEQ ID NOs: 14-26 and 39-44, or the L-nucleic acid molecule contains an L molecule -nucleic acid that is at least 75% identical to an L-nucleic acid molecule containing a nucleotide sequence selected from the group of SEQ NOs: 14-26 and 39-44, or the L-nucleic acid molecule contains an L-nucleic acid molecule that homologous to an L-nucleic acid molecule containing a nucleotide sequence selected from the group SEQ. ID NOs: 14-26 and 39-44, where such homology is at least 75%.

Option 10 : The L-nucleic acid molecule according to option 9, wherein the L-nucleic acid molecule contains a nucleotide sequence selected from the group SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 20 and SEQ ID NO: 41-44, or the L-nucleic acid molecule contains an L-nucleic acid molecule that is at least 75% identical to the L-nucleic acid molecule an acid containing a nucleotide sequence selected from the group SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 20 and SEQ ID NO: 41-44, or the L-nucleic acid molecule contains an L-nucleic acid molecule that is homologous to an L-nucleic acid molecule containing a nucleotide sequence selected from the group SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 20 and SEQ 41-44%, where the homology is at least 75%.

Option 11 : An L-nucleic acid molecule according to any one of options 1 to 10, wherein the L-nucleic acid molecule has the ability to bind to CXCL8, where preferably CXCL8 is human CXCL8, simian CXCL8, rabbit CXCL8, porcine CXCL8, canine CXCL8, ovine CXCL8 or CXCL8 guinea pig.

Option 12 : An L-nucleic acid molecule according to any one of embodiments 1 to 11, wherein the L-nucleic acid molecule has the ability to specifically bind to CXCL8, preferably human CXCL8, simian CXCL8, rabbit CXCL8, porcine CXCL8, canine CXCL8, ovine CXCL8 or CXCL8 guinea pig, and more preferably with human CXCL8.

Option 13 : An L-nucleic acid molecule according to any one of embodiments 1-12, wherein the L-nucleic acid molecule does not bind or is not capable of binding to ELR-positive CXC chemokines other than CXCL8, wherein the ELR-positive CXC chemokines other than CXCL8 , preferably selected from the group consisting of CXCL1, CXCL2, CXCL3, CXCL5, CXCL6 and CXCL7.

Option 14 : The L-nucleic acid molecule of embodiment 13, wherein the L-nucleic acid molecule does not bind or is not capable of binding to ELR-positive human CXC chemokines other than human CXCL8, wherein the ELR-positive human CXC chemokines other than human CXCL8 , are preferably selected from the group consisting of human CXCL1, human CXCL2, human CXCL3, human CXCL5, human CXCL6 and human CXCL7.

Option 15 : The L-nucleic acid molecule according to any one of embodiments 1 to 14, wherein the L-nucleic acid molecule has a binding affinity for human CXCL8, expressed as KD, of 10 nM or less, preferably 1 nM or less, more preferably 100 pM or less, and most preferably 30 pM or less.

Option 16 : The L-nucleic acid molecule according to any one of embodiments 1 to 15, wherein the L-nucleic acid molecule has a binding affinity for human CXCL8, expressed as IC ₅₀ , of 10 nM or less, preferably 1 nM or less, more preferably 100 pM or less, and most preferably 30 pM or less.

Option 17 : The L-nucleic acid molecule according to any one of embodiments 1 to 16, wherein the L-nucleic acid molecule has a binding affinity for ELR-positive chemokines CXC other than CXCL8 expressed as KD and being 100 nM or more, preferably 500 nM or more, and more preferably 1000 nM or more.

Option 18 : The L-nucleic acid molecule according to any one of embodiments 1 to 17, wherein the L-nucleic acid molecule has a binding affinity for ELR-positive chemokines CXC other than CXCL8 expressed as IC ₅₀ and being 100 nM or more, preferably 500 nM or more, and more preferably 1000 nM or more.

Option 19 : The L-nucleic acid molecule according to any one of embodiments 1 to 18, wherein the L-nucleic acid molecule has a binding affinity for human CXCL8, expressed as KD, of 10 nM or less, preferably 1 nM or less, more preferably 100 pM or less, and most preferably 30 pM or less, wherein the L-nucleic acid molecule has a binding affinity for ELR-positive chemokines CXC other than CXCL8, expressed as KD and being 100 nM or more, preferably 500 nM or more, and more preferably 1000 nM or more, and/or

the L-nucleic acid molecule has a binding affinity for human CXCL8, expressed as IC ₅₀ , of 10 nM or less, preferably 1 nM or less, more preferably 100 pM or less, and most preferably 30 pM or less, wherein the L-nucleic acid molecule has a binding affinity for ELR-positive chemokines CXC other than CXCL8, expressed as IC ₅₀ and being 100 nM or more, preferably 500 nM or more, and more preferably 1000 nM or more.

Embodiment 20 : The L-nucleic acid molecule according to any one of embodiments 15 to 19, wherein the binding affinity is determined at room temperature, preferably at 25°C.

Embodiment 21 : The L-nucleic acid molecule according to any one of embodiments 15 to 19, wherein the binding affinity is determined at 37°C.

Option 22 : An L-nucleic acid molecule according to any one of embodiments 1-21, wherein the L-nucleic acid molecule is an antagonist of activity mediated by CXCL8, preferably human CXCL8, simian CXCL8, rabbit CXCL8, porcine CXCL8, canine CXCL8, ovine CXCL8, or CXCL8 guinea pig, and more preferably human CXCL8.

Option 23 : The L-nucleic acid molecule of embodiment 22, wherein the L-nucleic acid molecule is not an antagonist of ELR-positive human CXC chemokines other than human CXCL8 or is not capable of inhibiting the activity mediated by ELR-positive CXC chemokines other than CXCL8, wherein the ELR-positive CXC chemokines other than CXCL8 are preferably selected from the group consisting of CXCL1, CXCL2, CXCL3, CXCL5, CXCL6 and CXCL7.

Option 24 : The L-nucleic acid molecule according to option 23, wherein the L-nucleic acid molecule is not an antagonist of ELR-positive human CXC chemokines other than human CXCL8, or is not capable of inhibiting the activity mediated by ELR-positive CXC chemokines other than CXCL8, wherein the ELR-positive human CXC chemokines other than human CXCL8 are preferably selected from the group consisting of human CXCL1, human CXCL2, human CXCL3, human CXCL5, human CXCL6 and human CXCL7.

Option 25 : The L-nucleic acid molecule according to any one of embodiments 22 to 24, wherein the antagonistic activity of the L-nucleic acid molecule to the activity mediated by human CXCR1 and/or CXCR2 and expressed as IC ₅₀ is 10 nM or less, preferably 1 nM or less, more preferably 100 pM or less, and most preferably 30 pM or less.

Embodiment 26 : The L-nucleic acid molecule according to any one of embodiments 22-25, wherein the antagonistic activity of the L-nucleic acid molecule to human CXCL8-mediated activity is determined at 37°C.

Embodiment 27 : An L-nucleic acid molecule according to any one of embodiments 1 to 26, wherein the L-nucleic acid molecule contains a modifying group.

Option 28 : The L-nucleic acid molecule according to option 27, where the modifying group is attached to the 5'-terminal nucleotide and/or 3'-terminal nucleotide of the L-nucleic acid molecule and/or to the nucleotide of the L-nucleic acid molecule between the 5'-terminal a nucleotide of an L-nucleic acid molecule and a 3'-terminal nucleotide of an L-nucleic acid molecule.

Embodiment 29 : The L-nucleic acid molecule according to any one of embodiments 27 and 28, wherein the modifying group is attached to the L-nucleic acid molecule via a linker.

Embodiment 30 : The L-nucleic acid molecule of embodiment 29, wherein the linker is a hydrophilic linker, wherein the hydrophilic linker preferably comprises one or more ethylene glycol units, and more preferably the hydrophilic linker comprises triethylene glycol, hexaethylene glycol or polyethylene glycol.

Embodiment 31 : The L-nucleic acid molecule of embodiment 29, wherein the linker is a biodegradable linker.

Embodiment 32 : The L-nucleic acid molecule according to any one of embodiments 27 to 31, wherein the rate of excretion of the L-nucleic acid molecule containing the modifying group from the body is reduced compared to the rate of excretion of the L-nucleic acid molecule not containing the modifying group.

Embodiment 33 : The L-nucleic acid molecule according to any one of embodiments 27 to 31, wherein the L-nucleic acid molecule containing the modifying group has a longer retention time in the body compared to the retention time of the L-nucleic acid molecule not containing the modifying group.

Embodiment 34 : The L-nucleic acid molecule according to any one of embodiments 32-33, wherein the organism is a human or animal organism, and preferably a human organism.

Option 35 : An L-nucleic acid molecule according to any one of embodiments 27 and 34, wherein the modifying group is selected from the group consisting of a biodegradable modification and a non-biodegradable modification, and wherein the modifying group is preferably selected from the group consisting of polyethylene glycol, straight chain polyethylene glycol, branched polyethylene glycol, hydroxyethyl starch, peptide, protein, polysaccharide, styrene, polyoxypropylene, polyoxyamidate and poly(2-hydroxyethyl)-L-glutamine.

Embodiment 36 : The L-nucleic acid molecule of embodiment 35, wherein the modifying group is a polyethylene glycol, preferably a straight chain polyethylene glycol or a branched polyethylene glycol, wherein the molecular weight of the polyethylene glycol is preferably from about 20,000 to about 120,000 Da, more preferably from about 30,000 to about 30,000 to about 80,000 Da, and most preferably about 40,000 Da.

Embodiment 37 : The L-nucleic acid molecule of embodiment 36, wherein the molecular weight of the polyethylene glycol is about 20,000 to about 120,000 Da, preferably about 30,000 to about 80,000 Da, and most preferably about 40,000 Da.

Embodiment 38 : The L-nucleic acid molecule of embodiment 35, wherein the modifying group is hydroxyethylated starch, wherein the molecular weight of the hydroxyethylated starch is preferably about 50 to about 1000 kDa, more preferably about 100 to about 700 kDa, and most preferably, from 200 to 500 kDa.

Embodiment 39 : An L-nucleic acid molecule according to any one of embodiments 27-31, wherein the modifying group is designed to immobilize the L-nucleic acid molecule.

Embodiment 40 : The L-nucleic acid molecule of embodiment 39, wherein the immobilization involves covalently linking the L-nucleic acid molecule to a surface, where the preferred surface is the surface of a reaction vessel, the surface of a device in the reaction vessel, or the surface of a biosensor.

Embodiment 41 : The L-nucleic acid molecule of embodiment 39, wherein the immobilization involves non-covalently binding the L-nucleic acid molecule to a surface, where the preferred surface is the surface of a reaction vessel, the surface of a device in the reaction vessel, or the surface of a biosensor.

Option 42 : An L-nucleic acid molecule according to any one of options 39-41, wherein the modifying group is selected from the group consisting of carboxyl, hydroxyl, phosphatidyl, sulfonic acid ester, amine, thiol, epoxide, alkyne, steric cycloalkyne, maleimide, azide, hydrazide, wherein the steric cycloalkyne is preferably selected from the group consisting of dibenzocyclooctyl (DBCO), bicyclo[6.1.0]non-4-yne (BCN) and azadibenzocyclooctyl (ADIBO).

Embodiment 43 : The L-nucleic acid molecule of embodiment 42, wherein the modifying group is a steric cycloalkyne, preferably dibenzocyclooctyl (DBCO).

Embodiment 44 : The L-nucleic acid molecule of Embodiment 41, wherein the modifying group is biotin, bromodeoxyuridine, digoxigenin or an oligonucleotide, and preferably the modifying group is biotin.

Embodiment 45 : An L-nucleic acid molecule according to any one of embodiments 27-31, wherein the modifying group allows the L-nucleic acid molecule to be detected, and preferably the modifying group is a label.

Option 46 : An L-nucleic acid molecule according to option 45, wherein the label is selected from the group consisting of biotin, bromodeoxyuridine, digoxigenin, oligonucleotide, fluorescent label, electrochemiluminescent label, radioactive label, enzyme label, UV label, colloidal gold, nanoparticle and label in form of a molecule forming a chelate complex.

Option 47 : An L-nucleic acid molecule according to any one of embodiments 1-46 for use in a method of treating and/or preventing a disease.

Option 48 : An L-nucleic acid molecule for use according to embodiment 47, wherein the disease is associated with a CXCL8-mediated pathogenic mechanism and/or is selected from the group consisting of an inflammatory disease and cancer, wherein the inflammatory disease is preferably an inflammatory disease of the respiratory tract, an inflammatory disease of the skin , autoimmune disease, inflammatory nerve disease, ischemic disease, atherosclerosis, pancreatic islet transplant rejection, organ transplant rejection, delayed organ function, spinal cord injury, or cystitis.

Embodiment 49 : An L-nucleic acid molecule according to any one of embodiments 1 to 47 for use in a CXCL8 detection method.

Embodiment 50 : An L-nucleic acid molecule according to any one of embodiments 1 to 47 for producing a detection agent or biosensor.

Embodiment 51 : A pharmaceutical composition comprising an L-nucleic acid molecule as defined in any one of embodiments 1-50, and optionally an additional component, wherein the additional component is selected from the group consisting of a pharmaceutically acceptable excipient, a pharmaceutically acceptable carrier, and a pharmaceutically active agent.

Embodiment 52 : The pharmaceutical composition of embodiment 51, wherein the pharmaceutical composition comprises an L-nucleic acid molecule as defined in any one of embodiments 1 to 50 and a pharmaceutically acceptable carrier.

Option 53 : Use of an L-nucleic acid molecule according to any one of options 1 to 50 to produce a medicament.

Option 54 : Use according to option 53, where the medicinal product is intended for human or veterinary use.

Option 55 : Use according to option 54, wherein the medicament is for the treatment and/or prevention of a disease that is associated with a CXCL8-mediated pathogenic mechanism and/or is selected from the group consisting of an inflammatory disease and cancer, wherein the inflammatory disease is preferably an inflammatory disease of the respiratory tract pathways, inflammatory skin disease, autoimmune disease, inflammatory nerve disease, ischemic disease, atherosclerosis, pancreatic islet graft rejection, organ transplant rejection, delayed organ function, spinal cord injury, or cystitis.

Option 56 : Use of an L-nucleic acid molecule according to any one of options 1-50 for the production of a diagnostic agent, diagnostic agent or biosensor.

Embodiment 57 : Use according to Embodiment 56, wherein the diagnostic agent or diagnostic agent has a surface on the reaction vessel or a surface on a device in the reaction vessel.

Option 58 : Application according to option 56, where the biosensor has a surface.

Embodiment 59 : Use as in any one of embodiments 56-58, wherein the diagnostic agent, diagnostic agent or biosensor is suitable for detecting, either directly or indirectly, CXCL8, and preferably human CXCL8.

Embodiment 60 : Use as in any one of embodiments 56-57, wherein CXCL8 is detected by direct binding, in a sandwich binding assay, or in a competitive binding assay.

Embodiment 61 : Use as in Embodiment 60, wherein direct binding includes a detection method that uses only the L-nucleic acid molecule of any one of Embodiments 1 to 50 and does not use or does not involve the use of a second molecule other than the L-nucleic acid molecule. a nucleic acid according to any of options 1-50 that interacts with CXCL8.

Embodiment 62 : Use as in Embodiment 60, wherein the sandwich assay is a sandwich hybridization assay in which an L-nucleic acid molecule according to any one of embodiments 1 to 50 is immobilized on a surface, and a complex of an L-nucleic acid molecule according to any one of embodiments 1 -50 and CXCL8 are detected using a detection agent that binds to the epitope(s) of CXCL8 that are different from the epitope(s) associated with the L-nucleic acid molecule according to any of embodiments 1-50 .

Embodiment 63 : Use as per Embodiment 60, wherein the sandwich assay is a sandwich hybridization assay, wherein an agent that binds to a CXCL8 epitope(s) that is different from the epitope(s) bound with an L-nucleic acid molecule according to any one of embodiments 1-50, is immobilized on a surface, and the complex of such agents and CXCL8 is detected using the L-nucleic acid molecule according to any one of embodiments 1-50.

Embodiment 64 : Use as in Embodiment 60, wherein in a competitive binding assay, an L-nucleic acid molecule according to any one of embodiments 1-50 is immobilized on a surface, and binding of the L-nucleic acid molecule according to any one of embodiments 1-50 to CXCL8 competes with oligonucleotides probes at least partially complementary to the L-nucleic acid molecule of any one of embodiments 1-50 or CXCL8, wherein the complementarity is preferably base complementarity, and more preferably is based on Watson-Crick base pairing.

Embodiment 65 : Use as in Embodiment 60, wherein in a competitive binding assay, the L-nucleic acid molecule according to any one of embodiments 1-50 is not immobilized on a surface, and the binding of the L-nucleic acid molecule according to any one of embodiments 1-50 to CXCL8 competes with oligonucleotide probes complementary to an L-nucleic acid molecule according to any one of embodiments 1-50 or CXCL8, where preferably, oligonucleotide probes complementary to an L-nucleic acid molecule according to any one of embodiments 1-50 or CXCL8 are immobilized on a surface, where the complementarity is preferably base complementarity, and more preferably, such complementarity is based on Watson-Crick base pairing.

Embodiment 66 : Use as in any one of embodiments 62-65, wherein an L-nucleic acid molecule according to any one of embodiments 39-44 is used for immobilization on the surface.

Option 67 : Use as in any one of Options 57-66, wherein the surface, and preferably the biosensor surface, is selected from the group consisting of gold, silver, titanium, zirconium, vanadium, chromium, manganese, cobalt, tungsten, molybdenum, platinum, aluminum, iron, steel, copper, nickel, silicon, germanium, indium phosphide, gallium arsenide and oxide, nitride or alloys or mixtures thereof, indium tin oxide, glassy carbon, sapphire, silicate glass and borate glass.

Embodiment 68 : Use according to any one of Embodiments 57-67, wherein the surface, preferably the surface of the biosensor, is modified with polylysine; aminosilane; epoxysilane; nitrocellulose; carboxydextran; carbon nanomembrane; graphene oxide; carbon surface such as black carbon, carbon fiber, carbon plate, carbon cloth, activated carbon, glassy carbon, carbon, activated carbon, graphite powder, graphite fiber, carbon nanotube; fullerene; carboxyl group; an azide group; thiol group; hydroxy group; epoxide; maleimide; alkyne; steric alkyne or any combination thereof.

Embodiment 69 : Use as in Embodiment 68, wherein to immobilize an L-nucleic acid molecule according to any one of Embodiments 39-44, the surface is functionalized with a primary amine, a thiol group, an azide, an alkyne, an alkene, a tetrazine, a tetrazole, or a steric cycloalkyne, wherein the steric cycloalkyne is preferably selected from the group consisting of dibenzocyclooctyl (DBCO), bicyclo[6.1.0]non-4-yne (BCN) or azadibenzocyclooctyl (ADIBO).

Embodiment 70 : Use as in Embodiment 69, wherein to immobilize the L-nucleic acid molecule according to any one of Embodiments 42-43, a steric alkyne-promoted azide-alkyne cycloaddition reaction is carried out.

Embodiment 71 : Use as in any one of embodiments 56 to 70, wherein the L-nucleic acid molecule according to any of embodiments 44 to 46 is used for detection.

Embodiment 72 : Use as in any one of Embodiments 56-71, wherein the biosensor detection system is based on optical sensing, mass change, refractive index, charge, surface tension, and interatomic force microscopy.

Embodiment 73 : Application as per Embodiment 72, where the biosensor detection system based on optical sensing is fluorescence, absorption or luminescence.

Option 74 : Application according to Option 72, where the biosensor detection system based on mass change is a quartz crystal microbalance system or microelectromechanical systems.

Option 75 : Application as per Option 72, where the refractive index based biosensor detection system is surface plasmon resonance, circular resonator or ellipsometry.

Embodiment 76 : Application as per Embodiment 72, where the charge-based biosensor detection system is electrochemical resistance, voltammetry, amperometry, potentiometry, conductance or field effect transistor.

Embodiment 77 : Application as per Embodiment 72, where the surface tension based biosensor detection system is a cantilever biosensor.

Embodiment 78 : Use as in any one of embodiments 56-59, wherein CXCL8 is detected using a homogeneity assay device.

Option 79 : Use of an L-nucleic acid molecule according to any one of embodiments 1-50 for detecting CXCL8, and preferably human CXCL8.

Embodiment 80 : Use as in Embodiment 79, wherein CXCL8 is detected using a sandwich binding assay, a homogeneity assay, or a competitive binding assay.

Embodiment 81 : Use as in Embodiment 80, wherein a sandwich binding assay, a homogeneity assay, or a competitive binding assay is suitable for direct or indirect detection of CXCL8.

Embodiment 82 : Use as in any one of Embodiments 78-81, wherein the L-nucleic acid molecule is covalently immobilized on the surface of a diagnostic reaction vessel, on the surface of a device in a diagnostic reaction vessel, or on the surface of a biosensor.

Embodiment 83 : Use as in Embodiment 82, wherein an L-nucleic acid molecule according to any one of Embodiments 39-44 is used for immobilization.

Option 84 : Use as in any one of Options 82-83, wherein the surface, and preferably the biosensor surface, is selected from the group consisting of gold, silver, titanium, zirconium, vanadium, chromium, manganese, cobalt, tungsten, molybdenum, platinum, aluminum, iron , steel, copper, nickel, silicon, germanium, indium phosphide, gallium arsenide and oxide, nitride or alloys or mixtures thereof, indium tin oxide, glassy carbon, sapphire, silicate glass and borate glass.

Option 85 : Use according to any of options 82-84, wherein the surface, and preferably the surface of the biosensor, is modified with polylysine; aminosilane; epoxysilane; nitrocellulose; carboxydextran; carbon nanomembrane; graphene oxide; carbon surface such as black carbon, carbon fiber, carbon plate, carbon cloth, activated carbon, glassy carbon, carbon, activated carbon, graphite powder, graphite fiber, carbon nanotube; fullerene; carboxyl group; an azide group; thiol group; hydroxy group; epoxide; maleimide; alkyne; steric alkyne or any combination thereof.

Embodiment 86 : Use as in Embodiment 85, wherein to immobilize an L-nucleic acid molecule according to any one of Embodiments 38-44, the surface is functionalized with an activated primary amine, a thiol group, an azide, or a steric cycloalkyne, wherein the steric cycloalkyne is preferably selected from the group consisting of dibenzocyclooctyl (DBCO ), bicyclo[6.1.0]non-4-yne (BCN) or azadibenzocyclooctine (ADIBO).

Embodiment 87 : Use as in Embodiment 86, wherein to immobilize the L-nucleic acid molecule according to any one of Embodiments 42-43, a steric alkyne-promoted azide-alkyne cycloaddition reaction is performed.

Embodiment 88 : Use as in any one of Embodiments 82 to 87, wherein the biosensor detection system is based on optical sensing, mass change, refractive index, charge, surface tension, and interatomic force microscopy.

Embodiment 89 : Application as per Embodiment 88, where the biosensor detection system based on optical sensing is fluorescence, absorption or luminescence.

Option 90 : Application according to Option 85, where the biosensor detection system based on mass change is a quartz crystal microbalance system or microelectromechanical systems.

Option 91 : Application as per Option 88, where the refractive index based biosensor detection system is surface plasmon resonance, circular resonator or ellipsometry.

Option 92 : Application as per Option 88, where the biosensor detection system based on charge is electrochemical resistance, voltammetry, amperometry, potentiometry, conductivity or field effect transistor.

Option 93 : Application according to Option 88, where the biosensor detection system based on surface tension is a cantilever biosensor.

Embodiment 94 : Use as in any one of Embodiments 79 to 81, wherein the L-nucleic acid molecule contains a modifying group that allows the L-nucleic acid molecule to be detected, and preferably the modifying group is a label.

Option 95 : Use as in Option 94, wherein the label is selected from the group consisting of biotin, bromodeoxyuridine, digoxigenin, oligonucleotide, fluorescent label, electrochemiluminescent label, radioactive label, enzyme label, UV label, colloidal gold, nanoparticle and molecule-forming label chelate complex.

Embodiment 96 : A kit for detecting CXCL8, wherein the kit comprises an L-nucleic acid molecule according to any one of embodiments 1-50 and at least an instruction insert or reaction vessel.

Option 97 : A method for detecting CXCL8 using an L-nucleic acid determined in any of embodiments 1-50 in a sample, wherein the method comprises the steps of:

a) obtaining a sample with an unknown concentration of CXCL8;

b) contacting the sample or diluting it with a diagnostic agent, diagnostic agent, or biosensor as defined in any of embodiments 56-78;

c) measuring a signal using a diagnostic agent, diagnostic agent, or biosensor as defined in any one of embodiments 56-78;

d) comparing, optionally, the signal with a reference;

where optionally, the concentration of CXCL8 in a sample with an unknown concentration of CXCL8 is determined by comparison with signals obtained from at least one sample with a known concentration of CXCL8, and preferably at least one sample with a known concentration of CXCL8 is subjected to steps b)-c).

Option 98 : A complex comprising an L-nucleic acid molecule according to any one of embodiments 1-50 and CXCL8, wherein the complex is preferably a crystalline complex.

Option 99 : A method of screening for an antagonist of CXCL8 mediated activity, wherein the method comprises the following steps:

- obtaining a candidate antagonist of CXCL8-mediated activity,

- obtaining an L-nucleic acid molecule defined in any of options 1-50;

- creating a test system that will transmit a signal in the presence of an antagonist of CXCL8-mediated activity, and

- confirming that the candidate antagonist of CXCL8-mediated activity is an antagonist of CXCL8-mediated activity.

Option 100 : A method of detecting an L-nucleic acid as defined in any of embodiments 1-50 in a sample, wherein said method comprises the steps of:

a) providing a capture probe, wherein the capture probe is at least partially complementary to the first portion of the L-nucleic acid molecule defined in any of embodiments 1-50, and a detection probe, wherein the detection probe is at least partially complementary to the second portion of the L molecule -nucleic acid defined in any of options 1-50; or, alternatively, wherein the capture probe is at least partially complementary to a second portion of the L-nucleic acid molecule as defined in any of embodiments 1-50 and the detection probe is at least partially complementary to the first portion of the L-nucleic acid molecule as defined in any one of embodiments 1-50. from options 1-50;

b) adding a capture probe and a detection probe, alone or in combination, to a sample containing an L-nucleic acid molecule defined in any of embodiments 1-50, or suspected of containing an L-nucleic acid molecule defined in any one of options 1-50;

c) causing the capture probe and the detection probe to interact, either simultaneously or sequentially in any order, with the L-nucleic acid molecule defined in any of embodiments 1-50, or a portion thereof;

d) optionally, determining whether the capture probe hybridizes with the L-nucleic acid molecule defined in any of options 1-50, as described in step a); And

e) detecting the complex obtained in step c) and consisting of an L-nucleic acid molecule defined in any of options 1-50, a capture probe and a detection probe.

Embodiment 101 : Method according to Embodiment 100, wherein the detection probe comprises a detection agent, and/or where the capture probe is immobilized on a support, and preferably a solid support.

Option 102 : The method according to option 100 or 101, wherein any detection probe that is not part of the complex obtained in step c) is removed from the reaction mixture so that only the detection probe that is part of the complex is detected in step e).

Embodiment 103 : The method according to any one of embodiments 100-102, wherein step e) includes the step of comparing the signal generated by the detection means if the capture probe and the detection probe hybridize in the presence of an L-nucleic acid molecule defined in any of embodiments 1-50 , or a portion thereof, and in the absence of said L-nucleic acid molecule as defined in any of embodiments 1-50.

Without being bound by any particular theory, the authors merely note that they have unexpectedly discovered that the biostable L-nucleic acid molecule of the invention binds specifically and with high affinity to human CXCL8, and thereby effectively inhibits the binding of CXCL8 to its receptors. The authors unexpectedly identified a nucleic acid molecule that specifically binds to CXCL8 and inhibits CXCL8, but does not bind to the related chemokines CXCL1, CXCL2, CXCL3, CXCL5, CXCL6 and CXCL7, and, accordingly, does not inhibit these chemokines, although these proteins have the same structural properties like CXCL8, such as the ELR motif. The reported affinity in the picomolar range and the high selectivity of nucleic acid binding to CXCL8 could not be predicted.

In particular, the present inventors have surprisingly found that the nucleic acid molecule of the invention is suitable for blocking the interaction of CXCL8 with its receptors and inhibits CXCL8-induced chemotaxis with an inhibition constant in the picomolar range. Thus, the nucleic acid molecule of the present invention can also be considered as an antagonist of the action of CXCL8, and in particular the action of CXCL8 on its receptors CXCR1 and/or CXCR2. Preferably, the CXCL8 antagonist used herein is a molecule that binds to CXCL8, such as the nucleic acid molecules of the invention, and inhibits the function of CXCL8, preferably in an in vitro assay as described in the examples, or in an in vivo model.

In accordance with the present invention, the nucleic acid molecule according to the invention is a nucleic acid. The terms “nucleic acid” and “nucleic acid molecule” are used interchangeably herein unless otherwise noted. Moreover, such nucleic acid(s) are preferably also referred to herein as nucleic acid molecule(s) of the present invention, nucleic acid(s) of the present invention, nucleic acid(s). acid(s) according to the invention, or nucleic acid molecule(s) according to the invention.

The features of the nucleic acids of the invention described herein may be implemented in any aspect of the present invention, including any embodiment thereof that uses a nucleic acid, either alone or in any combination. It should be noted that any variation of an aspect of the present invention is also a variation of each and every other aspect of the present invention. More specifically, any embodiment of the first aspect of the present invention is also an embodiment of each and any of the second, third, fourth, fifth, sixth, seventh, eighth, ninth, tenth, eleventh, twelfth and 13th aspects.

With respect to the various diseases, conditions and disorders that can be treated or prevented using the nucleic acid molecule of the invention and compositions, and preferably pharmaceutical compositions, containing these molecules, it should be noted that such diseases, conditions and disorders are preferably diseases , conditions and disorders described in this application, including, in particular, the diseases, conditions and disorders described and presented in the introductory part of this application. In addition, the relevant sections in the description and in the introductory part of the description constitute the main part of the description, which discusses the possible use of the nucleic acid molecule according to the invention for the prevention and treatment, respectively, of these diseases, conditions and disorders. In addition, the nucleic acid molecule of the invention is preferred if the physiological action of the CXCL8-CXCR1 receptor and the CXCL8-CXCR2 receptor system is associated with higher plasma levels of CXCL8.

As used herein, the term CXCL8 preferably means any CXCL8, including, but not limited to, mammalian CXCL8. Preferably, the mammalian CXCL8 is selected from the group consisting of human, monkey, rabbit, pig, dog, sheep, zebra fish and guinea pig CXCL8. More preferably, CXCL8 is human CXCL8, preferably having the amino acid sequence of SEQ ID No. 2.

As described in more detail in the claims and Example 1, the present inventors have unexpectedly identified a number of different nucleic acid molecules capable of binding to human CXCL8.

As described in more detail herein, the present inventors have identified a number of different CXCL8 nucleic acid molecules capable of binding to human CXCL8, wherein the nucleic acid molecules can be characterized as nucleotide fragments, which are also disclosed herein (see Example 1).

Each of the different types of CXCL8-binding nucleic acid molecules of the invention that bind to CXCL8 contains three different nucleotide fragments: a first terminal nucleotide fragment, a central nucleotide fragment and a second terminal nucleotide fragment. In general terms, a nucleic acid molecule binding to CXCL8 according to the invention contains at its 5' end and 3' end each of a terminal nucleotide fragment, that is, a first terminal nucleotide fragment and/or a second terminal nucleotide fragment (also called 5 '-terminal nucleotide fragment and 3'-terminal nucleotide fragment, respectively). The first terminal nucleotide fragment and the second terminal nucleotide fragment can, in principle, due to the complementarity of their bases, hybridize with each other, as a result of which a double-stranded structure will be formed during such hybridization. However, such hybridization does not necessarily occur in the molecule under physiological and/or non-physiological conditions. The three nucleotide fragments of the nucleic acid molecules binding to CXCCL8, namely, the first terminal nucleotide fragment, the central nucleotide fragment and the second terminal nucleotide fragment are located in relation to each other in the 5'→3' direction: the first terminal nucleotide fragment - the central nucleotide fragment - second terminal nucleotide fragment. Alternatively, the second terminal nucleotide fragment, the central nucleotide fragment and the first terminal nucleotide fragment are located with respect to each other in the 5'→3' direction.

The length of the central nucleotide fragments of the nucleic acids according to the invention is preferably 32 or 33 nucleotides.

The length of the first terminal nucleotide fragments of the nucleic acids according to the invention is from three to six nucleotides, preferably from five to six nucleotides, and more preferably five nucleotides.

The length of the second terminal nucleotide fragments of the nucleic acids according to the invention is from three to six nucleotides, preferably from five to six nucleotides, and more preferably five nucleotides.

As used herein, the terms “fragment” and “nucleotide fragment” are synonymous unless otherwise noted.

Differences in the sequence of certain fragments between different nucleic acid molecules that bind to CXCL8 may influence the binding affinity for CXCL8. Based on the binding assay of various nucleic acid molecules binding to CXCL8 according to the invention, the central fragment and the nucleotides forming this fragment, individually, and more preferably in their entire total length, play an important role in the binding of the CXCL8-binding nucleic acid molecule to CXCL8 .

In a preferred embodiment of the invention, the nucleic acid molecule of the invention is a single nucleic acid molecule. In a further embodiment of the invention, a single nucleic acid molecule is present as a plurality of single nucleic acid molecules or as a plurality of individual nucleic acid molecules of different species.

It will be apparent to those skilled in the art that the nucleic acid molecule of the invention preferably consists of nucleotides covalently linked to each other, preferably through phosphodiester bonds or linkers.

In accordance with the present invention, a nucleic acid molecule according to the invention contains two or more fragments or parts thereof, which, in principle, can hybridize with each other. After such hybridization, a double-stranded structure is formed. It will be apparent to those skilled in the art that such hybridization may or may not occur, particularly under in vitro and/or in vivo conditions. Moreover, in the case of hybridization, such hybridization, but not necessarily, occurs along the entire length of the two fragments, where, at least based on the rules of base pairing, in principle, such hybridization and thereby the formation of a double-stranded structure can occur. Preferably, the double-stranded structure used herein is part of a nucleic acid molecule or a structure formed by two or more separate strands or two spatially separated fragments of one strand of a nucleic acid molecule, where at least one, and preferably two or more base pairs are base pairs, and preferably bases pairing according to the Watson-Crick base pairing rules. It will also be apparent to those skilled in the art that other base pairings can occur, such as Hoogsteen base pairing, or that such pairing can produce a double-stranded structure. It should also be noted that such a feature of hybridization of two fragments preferably indicates that such hybridization is expected to occur due to base complementarity of the two fragments, regardless of whether such hybridization occurs in vivo and/or in vitro . In accordance with the present invention, such fragments are first terminal nucleotide fragments and second terminal nucleotide fragments, which, in one embodiment of the invention, can hybridize as defined above.

In a preferred embodiment of the invention, the term “arrangement” as used herein means the order or sequence of structural or functional features or elements described herein related to the nucleic acid molecule(s) disclosed herein.

The nucleic acid molecule of the invention also contains nucleic acids that are substantially homologous to the specific sequences disclosed herein. The term “substantially homologous” means that the homology is at least 75%, preferably 85%, more preferably 90%, and most preferably greater than 95%, 96%, 97%, 98% or 99%.

The actual percentage of homologous nucleotides present in the nucleic acid molecule of the invention will depend on the total number of nucleotides present in the nucleic acid molecule. The percentage change may be based on the total number of nucleotides present in the nucleic acid molecule.

Homology between two nucleic acid molecules can be determined by a method known to those skilled in the art. More specifically, a sequence comparison algorithm can be used to calculate the percentage homology of the test sequence(s) relative to the reference sequence based on specified program parameters. The test sequence is preferably a nucleic acid sequence or molecule that is considered homologous, or it can be tested for its homology and degree of homology with another nucleic acid molecule, and such other nucleic acid molecule is also called a reference sequence. In one embodiment of the invention, the reference sequence is a nucleic acid molecule described herein, and preferably a nucleic acid molecule having a sequence according to any of SEQ NO: 14-26 and 39-44, and preferably SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 25 and SEQ ID NO: 26, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 20 and SEQ ID NO: 41-44. Optimal sequence alignment for comparison can be carried out, for example, using the local homology algorithm according to Smith & Waterman (Smith & Waterman, 1981), using the Needleman & Wunsch homologous sequence alignment algorithm (Needleman & Wunsch, 1970), using the similarity search method according to Pearson and Lipman (Pearson & Lipman, 1988), using computerized implementations of these algorithms (GAP, BESTFIT, FASTA and TFASTA in the Wisconsin Genetics software package, Genetics Computer Group, 575 Science Dr. Madison, Wis.) or by visual observation .

One example of an algorithm that is suitable for determining percent sequence identity is the algorithm used in the basic local alignment search package (hereinafter referred to as “BLAST”), see, for example, Altschul Et Al. (Altschul et al. 1990 and Altschul et al. 1997). The computer program for conducting BLAST analyzes is publicly available on the website of the National Center for Biotechnology Information (hereinafter referred to as “NCBI”). The default parameters used to determine sequence identity using a computer program available at NCBI, such as BLASTN (for nucleotide sequences) and BLASTP (for amino acid sequences), are described by McGinnis et al. (McGinnis et al. 2004).

The nucleic acid molecule of the invention also contains a nucleic acid molecule that has a certain degree of identity to the nucleic acids disclosed and described herein and is determined by their nucleotide sequence. More preferably, the present invention also includes a nucleic acid molecule that is at least 75%, preferably 85%, more preferably 90%, and most preferably more than 95%, 96%, 97% 98% or 99 % identical to the nucleic acid molecule disclosed and described in this application and defined by its nucleotide sequence or part thereof.

The term “nucleic acid of the invention” or nucleic acid molecule of the invention also includes a nucleic acid molecule containing the nucleic acid sequences disclosed or described herein or a portion thereof, preferably to the extent that the nucleic acid molecule or portions are involved in binding to human CXCL8 . In one embodiment of the invention, such a nucleic acid is one of the nucleic acid molecules described or disclosed herein, or derivatives and/or metabolites thereof, wherein such derivative and/or metabolite is preferably a truncated nucleic acid molecule compared to nucleic acid molecules, described or disclosed in this application. Truncation may occur at either end or both ends of the nucleic acid molecules disclosed or described herein. In addition, truncation can be carried out in the internal nucleotide sequence of the nucleic acid molecule, that is, it can be carried out between the 5' and 3' terminal nucleotides, respectively. In addition, truncation may include deletion of at least one nucleotide in the sequence of nucleic acids described herein. Truncation may also be performed on more than one fragment of the nucleic acid(s) described or disclosed herein, where such fragment may be at least one nucleotide in length. The binding of a nucleic acid molecule according to the invention can be determined by one skilled in the art through routine experimentation or by using or employing the method described herein, and preferably described in the Examples section of this application.

The nucleic acid molecule according to the present invention may be either a D-nucleic acid molecule or an L-nucleic acid molecule. Preferably, the nucleic acid molecule according to the invention is an L-nucleic acid molecule.

Moreover, in one embodiment of the invention, each and every nucleic acid molecule described herein is included in its entirety within the scope of the term “nucleic acid sequence(s)”, but is not limited to the specifically identified nucleotide sequence(s). yami). In other words, the terms “comprising” or “comprises” are to be interpreted in this manner as “including” or “consisting of”.

Moreover, according to the present invention, a nucleic acid molecule of the invention is a portion of a longer nucleic acid molecule, wherein the longer nucleic acid comprises multiple portions, wherein at least one such portion is a nucleic acid of the invention or a portion thereof. The other portion(s) of such a longer nucleic acid molecule may be one or more D-nucleic acid molecules or one or more L-nucleic acid molecules. Any combination of these can be used in the present invention. These other portions of the longer nucleic acid molecule, taken alone or together, either in whole or in a particular combination, may have a function other than binding, and preferably binding, to CXCL8. One possible function is to interact with other molecules, and therefore such other molecules are preferably different from CXCL8, for example, in terms of immobilization, cross-linking, detection or amplification. In another embodiment of the invention, a nucleic acid molecule of the invention includes, as individual or combined fragments, multiple nucleic acid molecules of the invention. Such a nucleic acid molecule containing multiple nucleic acids according to the invention is also included within the scope of the term "longer nucleic acid".

As used herein, the term "L-nucleic acid" means a nucleic acid or nucleic acid molecule consisting of L-nucleotides, and preferably consisting entirely of L-nucleotides.

As used herein, the term “D-nucleic acid” means a nucleic acid or nucleic acid molecule consisting of D-nucleotides, and preferably consisting entirely of D-nucleotides.

As used herein, the terms “nucleic acid and nucleic acid molecule” are synonymous unless otherwise noted.

In addition, unless otherwise noted, any nucleotide sequence is presented herein in the 5'→3' direction.

Preferably, any nucleotide position used herein is defined or indicated relative to the 5' end of the sequence, fragment or subfragment containing such nucleotide. Accordingly, the second nucleotide is the second nucleotide when counted from the 5' end of the sequence, fragment and subfragment, respectively. Moreover, according to this, the penultimate nucleotide is the second nucleotide when counted from the 3' end of the sequence, fragment and subfragment, respectively.

The nucleic acid molecule according to the invention consists of ribonucleotides, where:

G is guanosine 5'-monophosphate,

C is cytidine 5'-monophosphate,

A is adenosine 5'-monophosphate,

U is uridine 5'-monophosphate.

To define ribonucleotide sequence motifs, the IUPAC abbreviations for ambiguous nucleotides are used:

S strong G or C W weak A or U(T) R purine G or A Y pyrimidine C or U (T) K keto G or U(T) M imino A or C B not A C or U (T) or G D not C A or G or U(T) H not G A or C or U (T) V not U A or C or G N All A or G or C or U(T)

Designing the nucleic acid molecule of the invention as an L-nucleic acid molecule is preferred for several reasons. L-nucleic acid molecules are enantiomers of naturally occurring nucleic acid molecules. However, D-nucleic acid molecules are not very stable in aqueous solutions, and in particular in biological systems or biological samples due to the presence of a wide range of nucleases. Naturally occurring nucleases, and in particular animal cell nucleases, are not capable of degrading L-nucleic acids. As a result, the biological half-life of the L-nucleic acid molecule increases significantly in such a system, including the animal and human body. Due to the lack of degradability of L-nucleic acid molecules, decomposition products by nucleases are not formed, and therefore no side effects are observed in such a system, including the animal and human body. This property allows L-nucleic acid molecules to be distinguished from virtually all other compounds that are used in the treatment of diseases and/or disorders associated with the presence of CXCL8. An L-nucleic acid molecule that specifically binds to a target molecule by a mechanism other than Watson-Crick base pairing, or an aptamer that consists partly or entirely of L-nucleotides, and in particular, a portion of an aptamer that participates in the binding of the aptamer to the molecule -target, also called optical isomer. Aptamers and optical isomers, as such, are known to those skilled in the art and, among others, are described in the Aptamer Handbook (eds. Klussmann, 2006).

According to the present invention, the nucleic acid molecule of the invention, whether present as a D-nucleic acid molecule, L-nucleic acid molecule or D, L-nucleic acid molecule, may be present as a single-stranded or double-stranded nucleic acid molecule acids. Typically, a nucleic acid molecule is a single-stranded nucleic acid molecule that has a certain secondary structure due to its primary sequence and can thus also form a tertiary structure. However, a nucleic acid molecule can also be double-stranded in the sense that two strands that are complementary or partially complementary to each other hybridize to each other.

The nucleic acid molecule of the invention may be modified. Such a modification can be made to a single nucleotide of a nucleic acid molecule and is well known to those skilled in the art. Examples of such modifications are described, among others, by Venkatesan et al. (Venkatesan et al. 2003) and Kusser (Kusser, 2000). Such a modification may be an H atom, an F atom, or an O-CH ₃ or NH ₂ group at the 2' position of one or more of all the individual nucleotides that make up the nucleic acid molecule. In addition, the nucleic acid molecule of the invention may contain at least one LNA nucleotide. In one embodiment of the invention, the nucleic acid molecule of the invention consists of LNA nucleotides.

In one embodiment, the nucleic acid molecule of the present invention may be a multicomponent nucleic acid molecule. As used herein, a multicomponent nucleic acid molecule is a nucleic acid molecule that consists of at least two separate nucleic acid chains. The at least two nucleic acid strands form a functional unit, wherein the functional unit is a ligand for a target molecule and preferably an antagonist of the target molecule, in this case CXCL8. At least two nucleic acid strands can be obtained from any nucleic acid molecule of the invention by cleavage of the nucleic acid molecule of the invention to form at least two strands or by synthesizing one nucleic acid molecule corresponding to the first part of the full-length nucleic acid molecule of the invention, and another nucleic acid molecule corresponding to another part of a full-length nucleic acid molecule according to the invention. Depending on the number of parts forming full-length nucleic acid molecules, a corresponding set of parts having the desired nucleotide sequence can be synthesized. It should be noted that the cleavage method and the synthesis method can be applied to obtain a multicomponent nucleic acid molecule where more than two chains are present, as described above. In other words, the at least two separate nucleic acid strands typically differ from the two strands that are complementary and hybridize with each other, although there may be a certain degree of complementarity between the at least two separate nucleic acid strands, and such complementarity may lead to hybridization of these individual chains.

Finally, the present invention also contemplates obtaining a fully closed, i.e., ring structure of the nucleic acid molecule of the invention, that is, in one embodiment of the invention, the nucleic acid molecule of the invention is closed, preferably through a covalent bond, where more preferably such a covalent bond is present between the 5' end and the 3' end of the nucleic acid sequence of a nucleic acid molecule according to the invention as described herein, or any derivative thereof.

Determination of the binding constants of a nucleic acid molecule according to the invention can be carried out using the methods described in Examples 3 and 4, which confirm that the nucleic acid molecule according to the invention has a suitable range of K _D values. A suitable measure for expressing the binding strength between a single nucleic acid molecule and a target, which in this case is CXCL8, is the so-called K _D value, which as such, as well as the method for determining it, are known to those skilled in the art.

Preferably, the K _D value for the nucleic acids according to the invention is below 1 μM. A K _D value of approximately 1 μM is considered characteristic of nonspecific binding of the nucleic acid to the target. As will be apparent to those skilled in the art, the K _D value for a group of compounds, such as the nucleic acid molecule of the invention, is within a certain range. The above-mentioned K _D value of approximately 1 μM is the preferred upper limit for the K _D value. The lower limit of the K _D for target-binding nucleic acids may be at least about 10 picomoles or higher. In accordance with the present invention, the K _D values for individual nucleic acids that bind to CXCL8 are preferably within this range. Preferred ranges can be determined by selecting any first number in that range and any second number in that range. Preferred upper K _D values are 250 nM, 100 nM and 20 nM, preferred lower K _D values are 10 nM or lower, 1 nM or lower, 100 pM or lower, and 30 pM or lower. A more preferred upper K _D value is 20 nM, and a more preferred lower K _D value is 30 pM or lower.

In addition to the binding properties of the nucleic acid molecule of the invention, the nucleic acid molecule of the present invention inhibits the function of the corresponding target molecule, which in this case is CXCL8. Inhibition of the function of CXCL8, for example, stimulation of the corresponding receptors, as described previously, is achieved by binding a nucleic acid molecule of the invention to CXCL8 to form a complex of the nucleic acid molecule of the invention and CXCL8. Such a complex of a nucleic acid molecule and CXCL8 cannot stimulate receptors that are normally stimulated by CXCL8, that is, CXCL8 that is not present in a complex with the nucleic acid molecule of the invention. Accordingly, the inhibition of receptor function by a nucleic acid molecule of the invention is independent of the corresponding receptor that can be stimulated by CXCL8, but results from preventing stimulation of the CXCL8 receptor by the nucleic acid molecule of the invention.

Determination of the inhibition constant for a nucleic acid molecule according to the invention can be carried out using the methods described in Example 5, which confirms the fact that the nucleic acid molecule according to the invention has a suitable inhibition constant allowing the use of said nucleic acids in a therapeutic treatment regimen. An appropriate measure for expressing the strength of the inhibitory effect on the interaction of a single nucleic acid molecule with the target, in this case CXCL8, and the corresponding receptor, is the so-called half-maximal inhibitory concentration (abbreviated IC ₅₀ ), which as such and the method for determining it are known to those skilled in the art. areas.

Preferably, the IC ₅₀ value for the nucleic acids according to the invention is below 1 μM. An IC ₅₀ value of approximately 1 μM is considered characteristic of nonspecific inhibition of target functions by a nucleic acid molecule. As will be apparent to those skilled in the art, the IC ₅₀ value for a group of compounds, such as the nucleic acid molecules of the invention, is within a certain range. The above-mentioned IC ₅₀ value of approximately 1 μM is the preferred upper limit for the IC ₅₀ value. The lower limit of the IC ₅₀ for a target-binding nucleic acid molecule may be at least about 10 picomoles or higher. In accordance with the present invention, the IC ₅₀ values for individual nucleic acids that bind to CXCL8 are preferably within this range. Preferred ranges can be determined by selecting any first number in that range and any second number in that range. Preferred upper IC ₅₀ values are 250 nM, 100 nM and 20 nM, preferred lower IC ₅₀ values are 10 nM or lower, 1 nM or lower, 500 pM or lower, and 260 pM or lower. A more preferred upper IC ₅₀ value is 20 nM, and a more preferred lower IC ₅₀ value is 260 pM or lower.

The nucleic acid molecule according to the invention can be of any length, provided that it has the ability to bind to a target molecule. It will be apparent to one skilled in the art that the nucleic acid molecules of the invention have a preferred length. Typically the length ranges from 15 to 120 nucleotides. It will be apparent to those skilled in the art that the possible length of a nucleic acid molecule according to the present invention is from 15 to 120 nucleotides. More preferred length ranges of the nucleic acid molecules of the invention are lengths ranging from about 20 to 100 nucleotides, about 20 to 80 nucleotides, about 20 to 60 nucleotides, about 38 to 45 nucleotides, and about 40 to 45 nucleotides.

In accordance with the present invention, the nucleic acid molecule according to the invention contains a part that is preferably a high molecular weight part and/or which preferably allows the properties of the nucleic acid molecule to be changed, where such properties include, among others, the residence time in the animal's body, and preferably in human body. A particularly preferred option for such modification is PEGylation and the introduction of a hydroxyethylated starch (HES) group into the nucleic acid molecule according to the invention. As used herein, PEG means polyethylene glycol and HES means hydroxyethyl starch. Preferably, PEGylation as used herein is a modification of a nucleic acid molecule of the invention, wherein the modification consists of a PEG molecule that is attached to a nucleic acid molecule of the invention. Preferably, the HES administration used herein is a modification of a nucleic acid molecule of the invention, wherein the modification consists of an HES molecule that is attached to a nucleic acid molecule of the invention. These modifications, as well as a method for modifying a nucleic acid molecule through such modifications, are described in patent applications EP1306382 and W02018099600A1, which are incorporated herein by reference in their entirety.

In the case of PEG, which is a high molecular weight group, the molecular weight is preferably from about 20,000 to about 120,000 Da, more preferably from about 30,000 to about 80,000 Da, and most preferably about 40,000 Da. In another preferred embodiment of the invention, the PEG is the PEG described in International Patent Application W003076490. In the case of HES being a high molecular weight group, the molecular weight is preferably from about 50 kDa to about 1000 kDa, more preferably from about 100 kDa to about 700 kDa, and most preferably from 200 kDa to 500 kDa. The molar substitution of HES is from 0.1 to 1.5, more preferably from 1 to 1.5, and such degree of substitution is expressed as a C2/C6 ratio of about 0.1 to 15, and preferably from about 3 to 10. A method for modifying HES is described, for example, in German patent application DE 142004006 249.8, which is incorporated herein by reference in its entirety.

In principle, a modification can be introduced into the nucleic acid molecule according to the invention at any position. Preferably, such a modification may be introduced into the 5' terminal nucleotide, the 3' terminal nucleotide, and/or any nucleotide between the 5' nucleotide and the 3' nucleotide of the nucleic acid molecule.

The modification, and preferably the PEG and/or HES group, can be attached to the nucleic acid molecule of the invention either directly or indirectly, preferably via a linker. Moreover, in accordance with the present invention, the nucleic acid molecule of the invention contains one or more modifications, and preferably one or more PEG and/or HES groups. In one embodiment of the invention, each linker molecule attaches more than one PEG group or HES group to the nucleic acid molecule of the invention. The linker used in the present invention may be straight or branched. Linkers of this type are known to those skilled in the art and are further described in International Patent Applications WO 2005/074993 and WO 2003/035665.

In a preferred embodiment of the invention, the linker is a biodegradable linker. The biodegradable linker allows the properties of the nucleic acid molecule according to the invention to be modified, where such properties include, among others, the residence time in the animal body, and preferably in the human body, due to the release of the modification from the nucleic acid molecule according to the invention. The use of a biodegradable linker allows for better control of the residence time of the nucleic acid molecule according to the invention. A preferred embodiment of such a biodegradable linker is the biodegradable linker described in International Patent Applications, including, but not limited to, WO 2006/052790, WO 2008/034122, WO 2004/092191 and W02005/099768.

In accordance with the present invention, the modification or modifying group is a biodegradable modification, where such biodegradable modification can be attached to the nucleic acid molecule of the invention either directly or indirectly, preferably through a linker. Biodegradable modification allows the properties of the nucleic acid molecule according to the invention to be changed, where such properties include, among others, the residence time in the animal body, and preferably in the human body, due to the release or decomposition of the modifying group from the nucleic acid molecule according to the invention. The use of a biodegradable modification allows for better control of the residence time of the nucleic acid molecule according to the invention. A preferred embodiment of such a biodegradable modification is the biodegradable modification described in International Patent Applications including, but not limited to, WO 2002/065963, WO 2003/070823, WO 2004/113394 and WO 2000/41647, and preferably WO 2000/41647, page 18, lines 4-24.

In another preferred embodiment of the invention, the linker has the following structure:

A suitable linker is, for example, described in WO 2015/062743. Based on the above structure, the linver connects two separate PEG molecules to the OH group of the phosphate portion of the nucleotide of the nucleic acid molecule according to the invention.

In addition to the modifications described above, other modifications may be used to change the properties of the nucleic acid molecule according to the invention, where such other modifications may be selected from the group of proteins, lipids such as cholesterol, and sugar chains such as amylase, dextran, etc. P.

Without being bound by any particular theory, we merely note that by modifying the nucleic acid molecule of the invention with a high molecular weight moiety such as a polymer, and more particularly one or more polymers described herein, which are preferably physiologically acceptable, the excretion kinetics of the so modified nucleic acid molecule according to the invention from an animal or human body to which a modified nucleic acid molecule according to the invention is administered changes. More specifically, due to the increase in molecular weight of the modified nucleic acid molecule according to the invention and due to the lack of metabolism of the nucleic acid molecule according to the invention, and especially if it is present in the L-Form, that is, in the form of an L-nucleic acid molecule, the rate of excretion from the body of the animal , preferably from a mammalian body, and more preferably from a human body, is reduced. Since excretion is usually via the kidneys, the present inventors anticipate that the rate of filtration of a nucleic acid molecule thus modified from the glomerulus is significantly reduced compared to a nucleic acid molecule not having this type of high molecular weight modification, resulting in an increased residence time of the modified nucleic acid molecule in the animal's body. In this regard, it is particularly noteworthy that, despite such high molecular weight modification, the specificity of the nucleic acid molecule according to the invention does not have a negative effect. Thus, the nucleic acid molecule of the invention, among other things, has a surprising property that is not typically observed for a pharmaceutically active compound, that is, the property that it is not necessary to use a sustained release pharmaceutical composition to provide sustained release of the nucleic acid molecule of the invention. Instead, the nucleic acid molecule of the invention in a modified form containing a high molecular weight group may be one that can already be used as a sustained release composition because it acts, by virtue of its modification, as if it were released from a sustained release composition . In this case, the modification(s) of the nucleic acid molecule of the invention as described herein, and the thereby modified nucleic acid molecule of the invention, and any composition containing these molecules, can provide distinct, preferably controlled pharmacokinetics and their biodistribution. This property also includes the residence time of these molecules in the bloodstream of the animal and human body and their distribution in the tissues of such animal and human. Such modifications are further described in patent application WO2003/035665.

However, in accordance with the present invention, the nucleic acid molecule according to the invention also does not contain any modification, and in particular does not have a high molecular weight modification such as PEG or HES. This embodiment of the invention is particularly preferred if the nucleic acid molecule of the invention has a preferential distribution in any organ or target tissue in the body, or if rapid clearance of the nucleic acid molecule of the invention from the body after its administration is desired. The nucleic acid molecule of the invention described herein, with a preferential distribution profile in any organ or target tissue in the body, allows effective local concentrations to be established in the target tissue while maintaining a low systemic concentration of the nucleic acid molecule. This allows the use of low doses, which are not only beneficial from an economic point of view, but also reduce the unwanted effects of the nucleic acid molecule on other tissues, and thereby reduce the potential risk of side effects. Rapid clearance of the nucleic acid molecule of the invention from the body after its administration may be desirable, among other things, in the case of in vivo imaging or obtaining specifically required therapeutic doses using the nucleic acid molecule of the invention or drugs containing such a molecule.

The nucleic acid according to the invention, which is also referred to herein as a nucleic acid according to the present invention or a nucleic acid molecule according to the present invention, and/or antagonists according to the present invention can be used for the production or preparation of a drug. Such a drug or pharmaceutical composition according to the invention contains at least one of the nucleic acids according to the invention, optionally together with other pharmaceutically active compounds, where the nucleic acid according to the invention preferably acts as the pharmaceutically active compound itself. In preferred embodiments of the present invention, such drugs contain at least a pharmaceutically acceptable carrier. Such a carrier may be, for example, water, buffer, PBS, glucose solution, preferably 5% glucose balanced salt solution, starch, sugar, gelatin or any other suitable carrier substance. Such carriers are essentially known to those skilled in the art. It will be apparent to one skilled in the art that any variations, uses, and aspects or related aspects relating to a drug of the invention are also applicable to the pharmaceutical composition of the invention and the nucleic acid molecule of the invention for the treatment and/or prevention and/or diagnosis of any of the disease described or disclosed herein, and vice versa.

The indications, diseases and disorders for the treatment and/or prevention of which the nucleic acids, pharmaceutical compositions and drugs in accordance with the present invention are used or obtained in accordance with the present invention are the result of the direct or indirect participation of CXCL8 in the corresponding pathogenesis.

CXCL8 (UniProtKB/Swiss-Prot PI 0145, IL8 HUMAN; SEQ ID NO: 2), also known as interleukin 8 (IL-8), neutrophil-activating protein 1 (NAP-1), monocyte neutrophil chemotaxis factor (MDNCDF) or Granulocyte chemotaxis protein 1 (GCP-1) is a small basic protein that belongs to the CXC subfamily of chemokines with proinflammatory and proangiogenic properties characterized by the presence of a glutamate-leucine-arginine (ELR) motif at the N-terminus. ELR-positive chemokines CXCL1 (also known as growth-regulatory protein alpha, Gro-alpha, protein with melanoma growth-promoting activity, MGSA or NAP-3), CXCL2 (also known as Gro-beta or macrophage inflammatory protein 2- alpha, MIP2-alpha), CXCL3 (also known as Gro-gamma or MIP2-beta), CXCL5 (also known as epithelial neutrophil-activating protein 78, ENA-78 or weakly inducible cytokine B5), CXCL6 (also known as chemokine alpha- 3, CKA-3, granulocyte chemotaxis protein 2, GCP-2, or weakly inducible cytokine B6), CXCL7 (also known as platelet basic protein, PBP, leukocyte growth factor, LDGF, macrophage growth factor, MDGF, or weakly inducible cytokine B7), and CXCL8 are agonists of the CXCR2 receptor (also known as IL8RB, IL8R type 2, CD182, CDwl28b or GRO/MGSA receptor). CXCL6, CXCL7 and CXCL8 are agonists of the CXCR1 receptor (also known as IL8RA, IL8R type 1, CD181 or CDwl28a). CXCL8 binds to the CXCR1 and CXCR2 receptors with similar affinities at a concentration of approximately 4 nM. Binding of CXCL8 to CXCR1/2 triggers Gαi-dependent signaling pathways and, for example, induces neutrophil migration, degranulation, and oxidative burst. Receptor sensitivity can be regulated through phosphorylation, beta-arrestin recruitment, and receptor internalization (Ha, Theranostics 2017). CXCL7 has the highest homology to CXCL8 with 33 identical amino acids. Non-human primate CXCL8 ( Macaca Mulata , a canine ape) is 95% identical (73 amino acids out of 77 amino acids are identical) to human CXCL8. There is no CXCL8 orthologue in mice and rats.

It will be appreciated that since the CXCL8-binding nucleic acid molecule of the invention interacts or binds with human CXCL8, one skilled in the art will appreciate that the CXCL8-binding nucleic acid molecule of the invention can be advantageously used for treatment, prophylaxis and/or diagnosis. any disease in humans and animals as described in this application. In this regard, it should be noted that the nucleic acid molecules of the invention can be used for the treatment and/or prevention of any disease, disorder or condition described herein, regardless of the mechanism of action underlying such disease, disorder or condition.

Further, without wishing to be bound by any particular theory, the authors merely note that the rational use of the nucleic acid molecules of the invention for the treatment of various diseases, disorders and conditions thereby provides the claimed therapeutic, prophylactic and diagnostic utility of the nucleic acid molecules of the invention. To avoid any unnecessary repetition, it should be noted that due to the inclusion of the CXCL8-CXCL8 receptor system described in this connection, the said system can serve as a target for the nucleic acid molecules according to the invention so that the claimed therapeutic, prophylactic and diagnostic effect is achieved. In addition, it should be noted that the characteristics of diseases, disorders and conditions in patients and any detailed description of the treatment regimen provided in connection therewith may be subject to invention related to preferred embodiments of the present invention.

CXCL8 and its receptors are or have been implicated in the pathophysiology of several inflammatory diseases, and the potential of CXCL8 inhibition to treat such inflammatory diseases is supported in animal models of such diseases. Inflammatory diseases and/or disorders and/or pathological conditions for the treatment and/or prevention of which a medicinal product according to the invention can be used may include, but are not limited to:

Inflammatory diseases of the respiratory tract:

A. Chronic obstructive pulmonary disease

b. Acute respiratory distress syndrome

With. Asthma and allergic inflammation

d. Bronchitis, bronchiolitis, bronchiolitis obliterans, bronchiectasis

e. Interstitial lung diseases, including pulmonary fibrosis

f. Cystic fibrosis

g. lung transplant

h. Lung damage caused by physical insults such as cigarette smoke, lung expansion after atelectasis, hyperoxia

i. Lung damage caused by a bacterial, viral, or fungal infection (pneumonia)

Inflammatory skin diseases:

A. Neutrophilic dermatoses

b. Psoriasis

With. Bullous pemphigoid

d. Epidermolysis bullosa

e. Hydraulitis suppurativa

f. Neurodermatitis

g. Eczema

Autoimmune diseases:

A. Inflammatory bowel diseases

b. Ulcerative colitis

With. Multiple sclerosis

d. Rheumatoid arthritis

e. Type 1 Diabetes

f. Ankylosing spondylitis

Nervous diseases:

A. Sweet's nervous disease

b. Alzheimer's disease

Ischemic reperfusion injury:

A. Stroke

b. Myocardial infarction

With. Ischemia and cerebral infarction

Other inflammatory diseases:

a. Atherosclerosis

b. Rejection of pancreatic islets

c. Rejection of transplanted organs and/or delayed function

d. Spinal cord injury

e. Cystitis

(Review provided by Russo, Expert Rev Clin Immunol 2014 and Ha, Theranostics 2017).

CXCL8 is overexpressed in human cancers, and experimental cancer models support the potential of CXCL8 inhibition for cancer treatment. Accordingly, diseases and/or disorders and/or pathological conditions for the treatment and/or prevention of which a medicinal product according to the invention can be used include, but are not limited to:

A. lungs' cancer

b. colon cancer

With. colorectal cancer

d. prostate cancer, including prostate cancer that is not treatable with hormones

e. pancreatic cancer

f. liver cancer

g. mammary cancer

h. ovarian cancer

i. thyroid cancer

j. melanoma

k. glioblastoma

l. osteosarcoma

m. esophageal carcinoma

n. blood cancer (leukemia and lymphoma)

o. PTEN-deficient tumors

p. p53-mutated tumors

q. Kras-mutated tumors

r. Treatment-resistant cancers, including cancers resistant to treatment with chemotherapeutic agents, antiangiogenic agents, tyrosine kinase inhibitors, and immune checkpoint inhibitors.

In another embodiment of the invention, the drug contains an additional pharmaceutically active agent for the treatment of cancer and/or tumors. Such additional pharmaceutically active compounds include, but are not limited to, compounds known to have antitumor actives such as alkylating agents, antimetabolites, antiangiogenic agents, mitosis inhibitory agents, topoisomerase inhibitors, cell signaling inhibitors, hormones, antibodies, immunoconjugates and hybrid proteins.

Other pharmaceutically active compounds for the treatment of cancer and/or tumors include immunotherapeutics. Such additional pharmaceutically active compounds include, but are not limited to, checkpoint inhibitors, cancer vaccines, immunostimulating agents, cellular agents, anticancer therapeutics.

Other pharmaceutically active compounds include, but are not limited to, compounds known as bleomycin, thymidylate synthase inhibitors such as raltitrexed and pemetrexed, enzymes such as L-asparaginase, miltefosine and angarelide, proteasome inhibitors such as bortezomib.

According to the present invention, a drug and a pharmaceutical composition, respectively, containing a nucleic acid according to the invention can be used for treatment in this manner.

In a further embodiment of the invention, the medicament contains an additional pharmaceutically active agent for the treatment of inflammatory diseases. Such additional pharmaceutically active compounds include, but are not limited to, compounds known to suppress the immune system such as non-steroidal anti-inflammatory drugs, corticosteroids, calcineurin inhibitors, cyclosporine A, methotrexate, azathioprine, tacrolimus, rapamycin, chlorambucil , leflunomide, mycophenolate mofetil, brequinar, mizoribine, thalidomide, deoxyspergualine, dapsone, hydroxychloroquine, sulfasalazine, antihistamines or anti-inflammatory biologics such as anti-tumor necrosis factor antibody, adalimumab, anti-CD20 B-cell depleting antibody rituximab, anti-IL6R tocilizumab antibody, anti-IgE antibody omalizumab and intravenous immunoglobulins.

Finally, the additional pharmaceutically active agent may be a modulator of the activity of any other chemokine, which may be a chemokine agonist or antagonist or a chemokine receptor agonist or antagonist. Alternatively or additionally, such additional pharmaceutically active agent is an additional nucleic acid molecule according to the invention. Alternatively, the drug contains at least one or more nucleic acids that bind to a target molecule other than CXCL8, or have a function different from the function of one of the nucleic acids according to the invention.

In accordance with the present invention, the drug is alternatively or additionally used, in principle, for the prevention of any of the diseases disclosed in connection with the use of the drug for the treatment of these diseases. Appropriate markers, that is, markers for identifying relevant diseases, are known to those skilled in the art. Preferably, the corresponding marker is CXCL8.

In one embodiment of a medicament according to the invention, such medicament is intended for use in combination with other methods of treating any of the diseases disclosed herein, and in particular, diseases for which the medicament according to the invention can be used.

"Combination therapy" (or "co-therapy") involves administering a medicament of the invention and at least a second agent as part of a specific treatment regimen to achieve the beneficial effect of the combined action of these therapeutic agents, that is, the medicament of the invention and said second agent . The beneficial effect of such a combination includes, but is not limited to, pharmacokinetic or pharmacodynamic synergies resulting from administration of the combination of therapeutic agents. Administration of these therapeutic agents in combination is usually carried out over a period of time (usually minutes, hours, days or weeks depending on the combination chosen).

"Combination therapy" may include, but typically does not include, the administration of two or more of these therapeutic agents as part of separate monotherapy regimens that randomly and arbitrarily result in the combination therapy of the invention. "Combination therapy" may include sequential administration of these therapeutic agents, that is, when each therapeutic agent is administered at a different time, as well as when these therapeutic agents or at least two of them are administered substantially simultaneously. Substantially simultaneous administration can be accomplished, for example, by administering to the individual a single capsule having a fixed ratio of each therapeutic agent or a plurality of separate capsules for each of the therapeutic agents.

Sequential or substantially simultaneous administration of each therapeutic agent can be accomplished by any suitable route, including, but not limited to, topical administration, oral administration, intravenous administration, intramuscular administration, and direct absorption through mucosal tissues. The therapeutic agents may be administered by the same route or by different routes. Thus, for example, the first therapeutic agent of the combination may be administered by injection, while the other therapeutic agents of the combination may be administered topically.

Alternatively, for example, all therapeutic agents may be administered topically, or all therapeutic agents may be administered by injection. The sequence of administration of therapeutic agents is not strictly defined unless otherwise noted. "Combination therapy" may also include the administration of the therapeutic agents described above in additional combination with other biologically active ingredients. If the combination therapy further includes non-drug treatment, then such non-drug treatment may be administered at any appropriate time until a beneficial effect from the combined action of the combination of therapeutic agents and non-drug treatment is achieved. Thus, for example, in some cases, a beneficial effect may still be achieved even if non-drug treatment is temporarily stopped when therapeutic agents are administered, perhaps for several days or even weeks.

As generally described above, the drug according to the invention can be administered in principle in any form known to those skilled in the art. The preferred route of administration is systemic administration, and more preferred is parenteral administration, preferably by injection. Alternatively, the drug may be administered topically. Other routes of administration include intramuscular, intraperitoneal and subcutaneous administration, oral administration, intranasal administration, intratracheal administration or pulmonary administration, with preference being given to the route of administration that is least invasive while still providing effectiveness.

Parenteral administration is usually used for subcutaneous, intramuscular or intravenous injections and infusions. In addition, one method of parenteral administration involves the implantation of sustained release or extended release systems, which will ensure that a constant dose level is maintained, as is well known to those skilled in the art.

In addition, preferred drugs of the invention may be administered intranasally by topical application of suitable intranasal vehicles, inhalers, or transdermally using transdermal skin patches in a form well known to those skilled in the art. When a transdermal delivery system is administered in this form, dosing is known to be continuous and uninterrupted throughout the dosing regimen. Other preferred topical preparations include creams, ointments, lotions, aerosol sprays and gels, in which the concentration of active ingredient is generally in the range of 0.01 to 15% w/w. or w/v

A medicament of the invention typically contains an effective amount of the therapeutic active component(s), including, but not limited to, a nucleic acid molecule of the invention, dissolved or dispersed in a pharmaceutically acceptable medium. Pharmaceutically acceptable vehicles or carriers include any and all solvents, dispersion media, coating agents, antibacterial and antifungal agents, isotonic and absorption retarding agents, and the like. The use of such media and agents for pharmaceutically active substances is well known to those skilled in the art. Additional active ingredients may also be included in the medicament according to the invention.

In another aspect, the present invention relates to a pharmaceutical composition. Such a pharmaceutical composition includes at least one of the nucleic acids according to the invention and preferably a pharmaceutically acceptable carrier. Such a carrier may be any carrier or any binding agent used and/or known to those skilled in the art. More specifically, such a binder or carrier is any binder or carrier as described in the formulation section of this application. In a further embodiment of the invention, the pharmaceutical composition contains an additional pharmaceutically active agent.

The preparation of the drug and pharmaceutical composition will be known to those skilled in the art from the description of the invention. Typically, such compositions may be prepared as injectable preparations as liquid solutions or suspensions; solid forms suitable for dissolution or suspension in liquid before injection; in the form of tablets or other solids for oral administration; in the form of delayed-release capsules; or in any other form currently in use, including eye drops, creams, lotions, ointments, inhalers, etc. It may also be particularly useful to use sterile compositions, such as saline washes, that are used by surgeons, physicians or medical personnel to treat a specific area of the surgical field. The compositions may also be delivered via a microdevice, microparticle or sponge.

Once formulated, the drug or pharmaceutical composition is administered in a manner compatible with the dosage form and in an amount that is pharmacologically effective. The compositions can be easily administered in various dosage forms, such as injection solutions of the type described above, but drug release capsules and the like can also be used.

In this context, the amount of active ingredient and the volume of the composition administered depends on the individual or patient being treated. The specific amounts of active compound required for administration depend on the judgment of the practitioner and are individual to each patient.

Typically, the minimum volume of drug or pharmaceutical composition necessary to disperse the active compounds is used. Suitable dosage regimens also vary but may be typical, for example, by initially administering the compound and monitoring results, and then additional controlled doses may be administered at additional intervals.

Thus, for example, for oral administration in the form of a tablet or capsule (eg, gelatin capsule), the active drug component, that is, the nucleic acid molecule of the invention and/or any other pharmaceutically active agent, also referred to herein as therapeutic agent(s) ) or active compound(s), may be combined with an oral, non-toxic, pharmaceutically acceptable inert carrier such as ethanol, glycerol, water and the like. In addition, if desired or necessary, suitable binders, lubricants, disintegrants and colorants may also be included in the mixture. Suitable binders are starch, magnesium aluminosilicate, starch paste, gelatin, methylcellulose, sodium carboxymethylcellulose and/or polyvinylpyrrolidone, natural sugars such as glucose or beta-lactose, corn sweeteners, natural and synthetic gums such as gum acacia, tragacanth or sodium alginate, polyethylene glycol, waxes, etc. Lubricants used in these dosage forms are sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride, silicon dioxide, talc, stearic acid, its magnesium or calcium salt and/or polyethylene glycol and the like. Disintegrants include, but are not limited to, starch, methylcellulose, agar, bentonite, starch-containing xanthan gum, agar, alginic acid or its sodium salt, or effervescent mixtures, and the like. Diluents are, for example, lactose, dextrose, sucrose, mannitol, sorbitol, cellulose and/or glycine.

The drug or pharmaceutical composition of the invention may also be administered in such oral dosage forms in the form of sustained and extended release tablets or capsules, dragees, powders, granules, elixirs, tinctures, suspensions, syrups and emulsions. Suppositories are mainly prepared from fat emulsions or suspensions.

The pharmaceutical composition or medicinal product may be sterilized and/or may contain adjuvants such as preservatives, stabilizers, wetting or emulsifying agents, dissolution promoters, salts to regulate osmotic pressure and/or buffers. In addition, they may also contain other therapeutically valuable substances. The compositions are prepared by standard mixing, granulating or coating processes and typically contain from about 0.1 to 75%, and preferably from about 1 to 50%, active ingredient.

Liquid compositions, and in particular injectable compositions, can be prepared, for example, by dissolving, dispersing, or the like. The active compound is dissolved or mixed with a pharmaceutically pure solvent such as, for example, water, saline, aqueous dextrose, glycerin, ethanol and the like. to form a solution or suspension for injection. In addition, solid forms suitable for dissolution in liquid prior to injection may be prepared.

For solid compositions, excipients are pharmaceutically pure substances such as mannitol, lactose, starch, magnesium stearate, sodium saccharin, talc, cellulose, glucose, sucrose, magnesium carbonate, etc. The active compound as defined above can also be prepared in the form of suppositories using, for example, polyalkylene glycols, eg propylene glycol, as a carrier. In some embodiments of the invention, suppositories are preferably prepared from fat emulsions or suspensions.

The drug, pharmaceutical composition and nucleic acid molecules of the invention, respectively, can also be administered in the form of liposomal delivery systems such as small unilamellar vesicles, large unilamellar vesicles and multilamellar vesicles. Liposomes can be formed from various phospholipids containing cholesterol, stearylamine or phosphatidylcholines. In some embodiments, the film of lipid components is hydrated with an aqueous drug solution to form a lipid layer encapsulating the drug, as is well known to those skilled in the art. Thus, for example, the nucleic acid molecules described herein can be obtained in the form of a complex with a lipophilic compound, or they can be obtained in the form of a non-immunogenic high molecular weight compound by methods known in the art. In addition, liposomes can carry such nucleic acid molecules on their surface to target and transport cytotoxic agents into the cell for cytolysis. An example of nucleic acid-associated complexes is presented in US Pat. No. 6011020.

The drug, pharmaceutical composition and nucleic acid molecules of the invention, respectively, can also be associated with soluble polymers as carriers for drug delivery. Such polymers may include polyvinylpyrrolidone, pyran copolymer, polyhydroxypropyl methacrylamide phenol, polyhydroxyethyl aspanamide phenol, or polyethylene oxide polylysine substituted with palmitoyl moieties. In addition, the drugs and nucleic acid molecules of the invention can accordingly be associated with specific classes of biodegradable polymers suitable for controlled drug release, such as, for example, polylactic acid, polyepsilon caprolactone, polyhydroxybutyric acid, polyorthoesters, polyacetals, polydihydropyrans, polycyanoacrylates and cross-linked or amphipathic block copolymers of hydrogels.

If desired, the pharmaceutical composition and drug administered, respectively, may also contain minor amounts of non-toxic excipients, such as wetting or emulsifying agents, pH buffering agents and other substances, such as, for example, sodium acetate and triethanolamine oleate.

The dosage regimen in which the nucleic acid molecule, drug or pharmaceutical composition of the invention is used is accordingly selected depending on a number of factors, including the type, species, age, weight, sex and health status of the patient; the severity of the condition being treated; method of administration; the patient's kidney and liver function; and the specific aptamer or salt thereof used. One skilled in the art, such as a physician or veterinarian, can readily determine and prescribe the effective amount of the drug needed to prevent, suppress, or stop the progression of the disease.

Effective plasma levels of the nucleic acid of the invention are preferably in the range of 500 fM to 500 μM for the treatment of any of the diseases described herein.

The nucleic acid molecule, drug and pharmaceutical composition according to the invention, respectively, can preferably be administered as a single daily dose, every second or third day, weekly, every two weeks, once a month or every three months.

In accordance with the present invention, the drug described herein is included in the pharmaceutical composition disclosed herein.

In a further aspect, the present invention relates to a method of treating an individual in need of such treatment, wherein said method comprises administering a pharmaceutically active amount of at least one of the nucleic acid molecules of the invention. In one embodiment of the invention, the individual suffers from a disease or is at risk of developing such a disease, wherein said disease is any of the diseases described herein, and in particular, any of the diseases disclosed herein in connection with the use of any nucleic acids according to the invention for the production of a drug.

It should be noted that the nucleic acid as well as the antagonists according to the invention can be used not only as a drug or as a drug preparation agent, but also for cosmetic purposes, and in particular for the treatment of inflammatory local skin lesions caused by CXCL8 . Therefore, an additional condition or disease that the nucleic acid, drug and/or pharmaceutical composition of the invention is used to treat or prevent is inflammatory localized skin lesions.

It will be appreciated that since the CXCL8-binding nucleic acid molecule of the invention interacts or binds to human CXCL8, one skilled in the art will appreciate that the CXCL8-binding nucleic acid molecule of the invention can be advantageously used to detect CXCL8 and to preparation of a diagnostic agent or biosensor.

Preferably, the diagnostic substance or diagnostic agent or diagnostic agent or biosensor used herein is suitable for detecting, either directly or indirectly, CXCL8, and preferably CXCL8 described in this application, and more preferably CXCL8 described in the section relating to the various disorders described herein and diseases. The diagnostic agent or biosensor is suitable for purposes such as, but not limited to, detecting and/or monitoring the progress of any of the disorders and diseases described herein, respectively. Such detection can be accomplished by binding the nucleic acid of the invention to CXCL8. Such binding can be detected either directly or indirectly, where the nucleic acid molecule may or may not contain a modifying group. In one preferred embodiment of the invention, the modifying group allows immobilization of the nucleic acid. In another preferred embodiment of the invention, the modifying group is a label. Suitable methods and means are known to those skilled in the art.

In one embodiment, the nucleic acid of the invention is immobilized on the surface of a diagnostic reaction vessel, on the surface of a device contained in a diagnostic reaction vessel, or on the surface of a biosensor, using any means known in the art, including non-covalent or covalent binding .

In a preferred embodiment of the invention, the nucleic acid according to the invention is immobilized on a surface by covalent binding. Thus, in the present invention, the nucleic acid molecule according to the invention contains a modifying group that ensures the immobilization of the nucleic acid molecule on surfaces. Such covalent linkage modifying groups include, but are not limited to, (activated) carboxyl, hydroxyl, phosphatidyl, sulfonic acid ester, amine, thiol, epoxide, alkyne, steric cycloalkyne (e.g. dibenzocyclooctyl, DBCO; bicyclo[6.1.0] non-4-yne, BCN or azadibenzocyclooctine, ADIBO), maleimide, azide, hydrazide.

Such covalent bonds can be formed, for example, by activated carboxyl groups (eg, through activation in the presence of EDC/NHS) reacting with primary amines; maleimide reacting with thiol groups via a copper-catalyzed azide-alkyne cycloaddition reaction (conducted by an automated chemistry method), using an automated copper-free chemistry method, including, but not limited to, copper-free azide-steric alkyne cycloaddition reaction, alkene cycloaddition reaction -azide [3+2], Diels-Alder alkene-tetrazine inversion reaction, and automated photo-alkene-tetrazole addition reaction, or through direct coupling of thiolated probes on gold surfaces. In this embodiment of the invention, such methods are not limited to a particular geometry, and the reactive group can be located on any surface or nucleic acids.

In a preferred embodiment of the invention, the nucleic acids of the invention are coupled to azide-functionalized biosensor surfaces via a sterically driven azide-alkyne cycloaddition reaction (automated chemistry). To do this, the surface is functionalized with an azide and the oligonucleotide is functionalized with a steric alkyne, for example, dibenzocyclooctyl, DBCO; bicyclo[6.1.0]non-4-yne (BCN) and azadibenzocyclooctyne (ADIBO), preferably at the 5' or 3' end. Alternatively, an inverted geometric structure can be used, that is, the reaction can be carried out on a biosensor surface functionalized with a steric alkyne and using azide-modified nucleic acids.

In another embodiment of the invention, the nucleic acid according to the invention is immobilized through non-covalent binding. Thus, in the present invention, the nucleic acid molecule according to the invention contains a modifying group that allows the nucleic acid molecule to be immobilized on surfaces where such binding is reversible. Such non-covalent binding modifying groups include, but are not limited to, biotin (binding to avidin, streptavidin, or neutravidin), bromodeoxyuridine (binding to an anti-bromodeoxyuridine antibody), digoxigenin (binding to an anti-digoxigenin antibody), and oligonucleotides (binding to a complementary oligonucleotide). The unblocked 3' ends of oligonucleotides can serve as primers for polymerases or as acceptors for another nucleic acid in a ligase reaction. In this embodiment of the invention, such methods are not limited to a particular geometry, and the reactive group can be located either on the surface or on the nucleic acids.

In the present invention, the nucleic acid molecule according to the invention is not immobilized, but is used in solution and contains a modifying group that preferably allows the detection of the nucleic acid molecule, herein referred to as a label. These tags include, but are not limited to, biotin (binding to avidin, streptavidin or neutravidin), bromodeoxyuridine (binding to anti-bromodeoxyuridine antibody), digoxigenin (binding to anti-digoxigenin antibody) and oligonucleotides (binding to complementary oligonucleotide), fluorescent tags (eg , fluorescein, Su-3, Su-5), electrochemiluminescent tags, radioactive tags, enzyme tags, UV tags, colloidal gold, nanoparticles and tags in the form of molecules that form chelate complexes.

In a further embodiment of the invention, the nucleic acid molecule of the invention contains a non-nucleoside, and preferably a hydrophilic linker, such as a single unit or repeating ethylene glycol, for example triethylene glycol (TEG), hexaethylene glycol (HEG) or even polyethylene glycol (PEG), which can link nucleic acid and modifying group.

With regard to the detection of CXCL8, the preferred methods are direct CXCL8 binding, sandwich binding assay, competitive binding assay or lateral flow device method.

In one embodiment of the invention, CXCL8 is detected by direct binding of immobilized nucleic acid to CXCL8. Direct binding according to the invention is a method in which only the nucleic acid according to the invention is used, but not a second molecule that interacts with CXCL8.

One format of a sandwich binding assay is the formation of an immobilized complex of CXCL8 and a nucleic acid molecule of the invention containing an immobilization modification group, where the complex is immobilized on the surface of a reaction vessel with a diagnostic agent, on the surface of a device contained in a reaction vessel with a diagnostic agent or on the surface of a biosensor with subsequent detection, preferably, of the specified complex. In one embodiment of the invention, CXCL8 is detected from the complex. A suitable detection agent that satisfies this requirement is, for example, any detection agent that is specific for that portion(s) of CXCL8 not detected by the nucleic acid molecule of the invention, and that is selected from the group consisting of nucleic acids, polypeptides, proteins and antibodies, the preparation of which is known to those skilled in the art. The technique for performing such sandwich binding assays is known to those skilled in the art.

In one embodiment, a sandwich binding assay may be performed that utilizes an inverted geometric structure where the detection agent described above is immobilized on the surface of a diagnostic reaction vessel, on the surface of a device contained in a diagnostic reaction vessel, or on surface of the biosensor with the formation of the CXCL8 complex in the sample, and subsequent detection of this complex by a nucleic acid molecule according to the invention. In this embodiment of the invention, the nucleic acid molecule of the invention contains a label or is unlabeled. Detection of an unlabeled nucleic acid molecule may involve the use of a second detection agent, which is preferably also selected from the group consisting of nucleic acids, polypeptides, proteins, and variants included in the various variants described herein. Such detection means are preferably specific for the nucleic acid of the invention.

Finally, in accordance with the present invention, the second detecting means is detected using a third detecting means, where preferably the third detecting means is associated with an enzyme, and more preferably, provides an enzymatic reaction after detection of the second detecting means, or where the third detecting means is a means for detecting radiation, and more preferably radiation emitted by a radionuclide. Preferably, the third detecting means allows the second detecting means to be specifically detected and/or interacting with the second detecting means.

In a further embodiment of the invention, the binding of nucleic acid molecules of the invention to CXCL8 is detected in competitive binding assays. In this case, immobilized oligonucleotide probes complementary to portions of the nucleic acid molecules of the invention compete to form a complex between the nucleic acid of the invention and CXCL8, where in such an embodiment of the invention, the nucleic acid molecule is not immobilized on the surface. Alternatively, immobilized CXCL8 competes for the formation of a complex between a nucleic acid of the invention and CXCL8, where in such an embodiment of the invention, the nucleic acid molecule is not immobilized on the surface. In a further embodiment of the invention, a nucleic acid molecule of the invention is immobilized on a surface, and binding of CXCL8 to the immobilized nucleic acid molecule of the invention competes with oligonucleotide probes complementary to portions of the nucleic acid molecules of the invention. The technique for such analyzes is known to those skilled in the art.

With regard to the detection of CXCL8, another preferred embodiment of the invention is the use of nucleic acid molecules according to the invention to obtain a biosensor. By definition, a biosensor is an analytical device used to detect an analyte, such as CXCL8, that combines a biological component, such as a nucleic acid of the invention, with a physicochemical detector. The transducer or detecting element, which converts one signal to another, operates by a physico-chemical mechanism, that is, optical, piezoelectric, electrochemical, electrochemiluminescent mechanism, etc., through the interaction of CXCL8 with a biological element. The biosensor may contain at least one biological element covalently immobilized on the surface of the biosensor. The at least one biological element covalently immobilized on the surface of the biosensor is a nucleic acid according to the invention containing a modifying group for immobilization, a means for specifically binding to this epitope/those epitopes of CXCL8 that are not associated with the nucleic acids according to the invention; an oligonucleotide probe complementary to the nucleic acids of the invention, or CXCL8. If the nucleic acid according to the invention is not immobilized on the surface of the biosensor, then the nucleic acid according to the invention contains a modifying group, which is a label, or does not contain a modifying group.

The biosensor surface or the diagnostic surface may be selected from the group consisting of gold, silver, titanium, zirconium, vanadium, chromium, manganese, cobalt, tungsten, molybdenum, platinum, aluminum, iron, steel, copper, nickel, silicon, germanium, indium phosphide, gallium arsenide and oxides, nitrides or alloys or mixtures of the above materials, indium tin oxide, glassy carbon, sapphire and silicate or borate glass.

These surfaces can be further modified with polylysine; aminosilane; epoxysilane; nitrocellulose; carboxydextran; carbon nanomembranes; graphene oxide; aminographene; carbon surface such as black carbon, carbon fiber, carbon plate, carbon cloth, activated carbon, glassy carbon, carbon, activated carbon, graphite powder, graphite fiber, carbon nanotube; fullerenes; carboxyl groups; azide groups; thiol groups; hydroxy groups; epoxide; maleimide; alkynes; steric alkynes or any combination thereof.

In a preferred embodiment of the invention, a nucleic acid molecule of the invention containing an immobilization modification group is immobilized on the surface of a biosensor for direct detection of CXCL8 or for detection of CXCL8 in a sandwich assay format or in a competitive binding assay format. In another preferred embodiment of the invention, a detection agent specific for this CXCL8 epitope/epitopes not detected by the nucleic acid according to the invention (described above); an oligonucleotide probe complementary to the nucleic acids according to the invention or CXCL8, which were immobilized on the surface of the biosensor; and labeled or unlabeled nucleic acids according to the invention (described above) are used for detection in a sandwich assay format or in a competitive binding assay format.

Detection systems in diagnostics and biosensors can be based, for example, on optical sensing (fluorescence, absorption and luminescence), on mass change (for example, microbalance based on quartz crystals and microelectromechanical systems), on refractive index (based on surface plasmon resonance, circular resonator, ellipsometry), charge (based on electrochemical resistance, voltammetry, amperometry, potentiometry, conductivity or field-effect transistors), surface tension (based on cantilever biosensors) and atomic force microscopy.

In a further embodiment of the invention, the nuleic acid molecule of the invention is used in sandwich or lateral flow competition binding assays. The technique of such lateral flow assays is known to those skilled in the art.

In a further embodiment of the invention, binding of a nucleic acid molecule of the invention to CXCL8 is detected in homogeneity assays using nucleic acid probes complementary to portions of the nucleic acid molecule of the invention, where in such an embodiment the nucleic acid molecule is not immobilized on a surface. Here, hybridization results in changes in the fluorescent signal of the two fluorophore groups associated with the nucleic acids, preferably a change in intensity, and such hybridization competes for the formation of a complex between the nucleic acid of the invention and CXCL8. The two fluorophore groups are either both bound to the nucleic acid probe (molecular beacon), or one of them is bound to the probe and the other is bound to the nucleic acid of the invention. In a further embodiment of the invention, one or two fluorophore groups are associated with a nucleic acid according to the invention, and formation of a complex with CXCL8 results in changes in the fluorescent signal, and preferably a change in intensity. The technique for such analyzes is known to those skilled in the art.

The nucleic acid molecule of the invention can also be used as a starting material for drug development. Basically, there are two possible approaches. One such approach is the screening of compound libraries, where each compound library is preferably a library of small molecule compounds. In one embodiment of the invention, the screening is a large-scale screening. Preferably, large-scale screening is a rapid, efficient trial-and-error assessment of compounds in a target-based assay. In the best case scenario, assays are performed by colorimetric evaluation. Libraries used in such screening are known to those skilled in the art.

Alternatively, the nucleic acid molecule of the invention can be used for rational drug development. Preferably, rational drug development is the creation of a basic pharmaceutical structure. Based on the three-dimensional structure of the target, which is usually identified by methods such as X-ray crystallography or nuclear magnetic resonance spectroscopy, computer programs are used to search databases containing the structures of many different chemical compounds. Selection is carried out on a computer, and identified compounds can subsequently be tested in the laboratory.

Rational drug development can begin with any nucleic acid molecule of the invention and involves searching for a structure, preferably a three-dimensional structure, that is similar to the structure of the nucleic acids of the invention or identical to the binding mediating portions of the nucleic acid structure of the invention. In any case, such a structure will still exhibit the same binding property as the nucleic acid of the invention, or a similar binding property. As a next step, or as an alternative step in rational drug development, it is preferable that the three-dimensional structure of these neurotransmitter-binding nucleic acid portions be masked by chemical groups that are different from nucleotides and nucleic acids. By means of such mimicry, a compound different from the nucleic acid according to the invention can be obtained. Such a compound is preferably a small molecule or peptide.

By screening libraries of compounds, for example using a competitive binding assay known to one skilled in the art, corresponding CXCL8 analogs, CXCL8 agonists or CXCL8 antagonists can be found. Such competitive binding assays can be performed as follows. The nucleic acid according to the invention, preferably the optical isomer, which is a target-binding L-nucleic acid, is attached to the solid phase. To identify CXCL8 analogues, tagged CXCL8 is added to the assay. The potential analog will compete with CXCL8 molecules for binding to the optical isomer, which will be accompanied by a decrease in the signal transmitted by the corresponding label. Screening for agonists or antagonists may involve the use of cell culture assays known to those skilled in the art.

The kit according to the invention may contain at least one or more nucleic acid molecules according to the invention. In addition, the kit may contain at least one or more positive or negative controls. A positive control can be, for example, CXCL8, and in particular CXCL8, against which the nucleic acid according to the invention is selected or to which it binds preferably in liquid form. The negative control may be, for example, a peptide that has biophysical properties similar to those of CXCL8, but which is not recognized by the nucleic acids of the invention. In addition, the specified set may contain one or more buffers. The various ingredients may be contained in the kit in dried or lyophilized form, or they may be dissolved in liquid. A kit may contain one or more containers, which in turn may contain one or more kit ingredients. In a further embodiment of the invention, the kit contains instructions or an instruction insert that provides the user with information on how to use the kit and its various ingredients.

The pharmaceutical and bioanalytical determination of a nucleic acid molecule according to the invention is an elemental analysis for assessing its pharmacokinetic and biodynamic profile in various physiological fluids, tissues and organs of the human and animal body. For this purpose, any of the detection methods disclosed in this application or known to a person skilled in the art can be used. In a further aspect of the present invention, a sandwich hybridization assay is used to detect the nucleic acid of the invention. The detection assay uses a capture probe and a detection probe. The capture probe is complementary to the first part and the detection probe is complementary to the second part of the nucleic acid according to the invention. The capture probe and detection probe may be formed by DNA nucleotides, modified DNA nucleotides, modified RNA nucleotides, RNA nucleotides, LNA nucleotides and/or PNA nucleotides.

Thus, the capture probe contains a sequence fragment complementary to the 5' end of the nucleic acid molecule of the invention, and the detection probe contains a sequence fragment complementary to the 3' end of the nucleic acid molecule of the invention. In this case, the capture probe is immobilized on the surface or matrix at its 5' end, whereby the capture probe can be immobilized directly at its 5' end or through a linker located between its 5' end and the surface or matrix. However, in principle, a linker can be associated with each nucleotide of the capture probe. The linker can be formed by hydrophilic linkers known to those skilled in the art, or D-DNA nucleotides, modified D-DNA nucleotides, D-RNA nucleotides, modified D-RNA nucleotides, D-LNA nucleotides, PNA nucleotides, L-RNA nucleotides, L-DNA, modified L-RNA nucleotides, modified L-DNA nucleotides and/or L-LNA nucleotides.

Alternatively, the capture probe contains a sequence fragment complementary to the 3' end of the nucleic acid molecule of the invention, and the detection probe contains a sequence fragment complementary to the 5' end of the nucleic acid molecule of the invention. In this case, the capture probe is immobilized on the surface or matrix at its 3' end, whereby the capture probe can be immobilized directly at its 3' end or through a linker located between its 3' end and the surface or matrix. However, in principle, a linker can be associated with each nucleotide of a sequence fragment that is complementary to a nucleic acid according to the invention. The linker can be formed by hydrophilic linkers known to those skilled in the art, or D-DNA nucleotides, modified D-DNA nucleotides, D-RNA nucleotides, modified D-RNA nucleotides, D-LNA nucleotides, PNA nucleotides, L-RNA nucleotides, L-DNA, modified L-RNA nucleotides, modified L-DNA nucleotides and/or L-LNA nucleotides.

The number of nucleotides of the capture probe and detection probe that can hybridize with a nucleic acid molecule of the invention may be variable and may depend on the number of nucleotides of the capture probe and/or detection probe and/or the nucleic acid of the invention itself. The total number of nucleotides of the capture probe and detection probe that can hybridize with the nucleic acid of the invention should represent the maximum number of nucleotides that are composed of the nucleic acid of the invention. The minimum number of nucleotides (from 2 to 10 nucleotides) of the detection probe and the capture probe must ensure hybridization to the 5' end or 3' end, respectively, of the nucleic acid according to the invention. To ensure high specificity and selectivity between the nucleic acid of the invention and other nucleic acids present in the samples that have been analyzed, the total number of nucleotides of the capture probe and detection probe should represent the maximum number of nucleotides that comprise the nucleic acid molecule of the invention.

In addition, the detection probe preferably carries a marker molecule or label that can be detected as described above in this application. A label or marker molecule can, in principle, be associated with each nucleotide of the detection probe. Preferably, the tag or marker is located at the 5' end or 3' end of the detection probe, whereby a linker can be inserted between nucleotides within the detection probe that are complementary to the nucleic acid molecule of the invention and the tag. The linker can be formed by hydrophilic linkers known to those skilled in the art, or D-DNA nucleotides, modified D-DNA nucleotides, D-RNA nucleotides, modified D-RNA nucleotides, D-LNA nucleotides, PNA nucleotides, L-RNA nucleotides, L-DNA modified with L-RNA nucleotides, modified L-DNA nucleotides and/or L-LNA nucleotides.

The detection of nucleic acid according to the invention can be carried out as follows.

The nucleic acid molecule according to the invention is hybridized at one of its ends with a capture probe and at the other end with a detection probe. Thereafter, the unbound detection probe is removed, for example by one or more washing steps. The amount of bound detection probe, which preferably carries a label or marker molecule, can subsequently be estimated, for example, as described in more detail in WO/2008/052774, which is incorporated herein by reference.

Preferably, as used herein, the term “treatment” includes, in a preferred embodiment of the invention, additionally or alternatively, prophylaxis and/or monitoring.

Preferably, the terms “disease” and “disorder” as used herein are synonymous unless otherwise noted.

As used herein, the term “includes” is preferably not intended to limit the subject matter that is followed by or described by such term. However, in an alternative embodiment of the invention, the term “includes” is to be understood as “comprises” and, thus, is considered to be a limitation of the subject matter followed or described by such term.

The various sequences of SEQ ID NO:, the chemical nature of the nucleic acid molecules of the invention and the CXCL8 target molecules used in the present application, their actual sequence and internal reference standard are arranged in the following table.

Table 1

SEQ. ID. NO: Name Sequence (N-terminus → C-terminus) 1 D-amino acids D-CXCL8
(C-bio)(also called "biotinylated D-CXCL8" herein) AVLPRSAKELRCQCIKTYSKPFHPKFIKELRVIESGPHCAN
TEIIVKLSDGRELCLDPKENWVQRVVEKFLKRAENS-biotin 2 L-amino acids CXCL8, also called "L-CXCL8") AVLPRSAKELRCQCIKTYSKPFHPKFIKELRVIESGPHC
ANTEIIVKLSDGRELCLDPKENWVQRVVEKFLKRAENS 3 D-RNA 315-H9-001 GCUGACGGAAGUACGUGGAAAGCCAAUGAGUGUGUCCCGGUCAGC 4 D-RNA 315-F11-001 GCUGACGGAAGUACGUGGAAAGCCGAUGAGUGUGUCCCGGUGAGC 5 D-RNA 315-F8-001 GCUGACGGAAGUACGUGGAAAGCCGAAAGUGUGUCCCGGUCAGC 6 D-RNA 315-F8-002 CUGACGGAAGUACGUGGAAAGCCGAAAGUGUGUCCCGGUCAG 7 D-RNA 315-F8-003 UGACGGAAGUACGUGGAAAGCCGAAAGUGUGUCCCGGUCA 8 D-RNA 315-F8-004 GACGGAAGUACGUGGAAAGCCGAAAGUGUGUCCCGGUC 9 D-RNA 315-F8-005 GUGACGGAAGUACGUGGAAAGCCGAAAGUGUGUCCCGGUCAC 10 D-RNA 315-F8-006 GGACGGAAGUACGUGGAAAGCCGAAAGUGUGUCCCGGUCC eleven D-RNA 315-F8-007 GCACGGAAGUACGUGGAAAGCCGAAAGUGUGUCCCGGUGC 12 D-RNA 315-F8-008 GGUCGGAAGUACGUGGAAAGCCGAAAGUGUGUCCCGGACC 13 D-RNA 315-F8-009 UGGCGGAAGUACGUGGAAAGCCGAAAGUGUGUCCCGGCCA 14 L-RNA 315-H9-001 GCUGACGGAAGUACGUGGAAAGCCAAUGAGUGUGUCCCGGUCAGC 15 L-RNA 315-F11-001 GCUGACGGAAGUACGUGGAAAGCCGAUGAGUGUGUCCCGGUGAGC 16 L-RNA 315-F8-001 GCUGACGGAAGUACGUGGAAAGCCGAAAGUGUGUCCCGGUCAGC 17 L-RNA 315-F8-002 CUGACGGAAGUACGUGGAAAGCCGAAAGUGUGUCCCGGUCAG 18 L-RNA 315-F8-003 UGACGGAAGUACGUGGAAAGCCGAAAGUGUGUCCCGGUCA 19 L-RNA 315-F8-004 GACGGAAGUACGUGGAAAGCCGAAAGUGUGUCCCGGUC 20 L-RNA 315-F8-005 GUGACGGAAGUACGUGGAAAGCCGAAAGUGUGUCCCGGUCAC 21 L-RNA 315-F8-006 GGACGGAAGUACGUGGAAAGCCGAAAGUGUGUCCCGGUCC 22 L-RNA 315-F8-007 GCACGGAAGUACGUGGAAAGCCGAAAGUGUGUCCCGGUGC 23 L-RNA 315-F8-008 GGUCGGAAGUACGUGGAAAGCCGAAAGUGUGUCCCGGACC 24 L-RNA 315-F8-009 UGGCGGAAGUACGUGGAAAGCCGAAAGUGUGUCCCGGCCA 25 L-RNA 315-F8-002-PEG, 315-F8-002-5'-PEG PEG linker-CUGACGGAAGUACGUGGAAAGCCGAAAGUGUGUCCCGGUCAG; linker=aminohexyl 26 L-RNA 315-F8-002-amino, 315-F8-002-5'-amino NH2-linker-CUGACGGAAGUACGUGGAAAGCCGAAAGUGUGUCCCGGUCAG; 27 GGAAGUACGUGGAAAGCCRA(XU)RAGUGUGUCCCG
where X _U is U or absent 28 GGAAGUACGUGGAAAGCCAAUGAGUGUGUCCCG 29 GGAAGUACGUGGAAAGCCGAUGAGUGUGUCCCG thirty GGAAGUACGUGGAAAGCCGAAAGUGUGUCCCG 31 Reverse 315-F8-002-amino NH2-linker-GACUGGCCCUGUGUGAAAGCCGAAAGGUGCAUGAAGGCAGUC 32 L-amino acids CXCL1 ASVATELRCQCLQTLQGIHPKNIQSVNVKSPGPHCAQ
TEVIATLKNGRKACLNPASPIVKKIIEKMLNSDKSN 33 L-amino acids CXCL2 APLATELRCQCLQTLQGIHLKNIQSVKVKSPGPHCAQTE
VIATLKNGQKACLNPASPMVKKIIEKMLKNGKSN 34 L-amino acids CXCL3 ASVVTELRCQCLQTLQGIHLKNIQSVNVRSPGPHCAQTE
VIATLKNGKKACLNPASMVQKIIEKILNKGSTN 35 L-amino acids CXCL5 AGPAAAVLRELRCVCLQTTQGVHPKMISNLQVFAIGPQCS
KVEVVASLKNGKEICLDPEAPFLKKVIQKILDGGNKEN 36 L-amino acids CXCL6 GPVSAVLTELRCTCLRVTLRVNPKTIGKLQVFPAGPQCSK
VEVVASLKNGKQVCLDPEAPFLKKVIQKILDSGNKKN 37 L-amino acids CXCL7 SSTKGQTKRNLAKGKEESLDSDLYAELRCMCIKTTSGIHPKNIQSLEV
IGKGTHCNQVEVIATLKDGRKICLDPDAPRIKKIVQKKLAGDESAD 38 L-amino acids CCL5 SPYSSDTTPCCFAYIARPLPRAHIKEYFYTSGKCSNPAVVFVTRKNRQVCANPEKKWVREYINSLEMS 39 L-RNA 315-F8-002-3'-PEG CUGACGGAAGUACGUGGAAAGCCGAAAGUGUGUCCCGGUCAG-linker-PEG; linker=aminohexyl 40 L-RNA 315-F8-002-3'-amino CUGACGGAAGUACGUGGAAAGCCGAAAGUGUGUCCCGGUCAG-linker-amino; linker=aminohexyl 41 L-RNA 315-F8-005-5'-PEG PEG linker-GUGACGGAAGUACGUGGAAAGCCGAAAGUGUGUCCCGGUCAC;
linker=aminohexyl 42 L-RNA 315-F8-005-5'- amino NH2-linker-GUGACGGAAGUACGUGGAAAGCCGAAAGUGUGUCCCGGUCAC;
linker=aminohexyl 43 L-RNA 315-F8-005-3'-PEG GUGACGGAAGUACGUGGAAAGCCGAAAGUGUGUCCCGGUCAC-linker-PEG; linker=aminohexyl 44 L-RNA 315-F8-005-3-amino GUGACGGAAGUACGUGGAAAGCCGAAAGUGUGUCCCGGUCAC-linker-amino; linker=aminohexyl 45 D-RNA 8A-35
(Sung, Biomaterials 2014) 5'-GGGGGCUUAUCAUUCCAUUUAGUGUUAUGAUAACC-3'
C=2-fluoro-C; U=2'-fluorine-U PEG=Polyethylene glycol 40 kDa in Y-form

The features of the present invention, disclosed in the description, in the claims, in the sequence listing and/or in the drawings, can, either individually or in any combination thereof, serve as material for implementing the invention in various forms.

The present invention is further illustrated by drawings, examples and a sequence listing from which additional features, variations and advantages may be apparent, wherein:

In fig. 1 shows a sequence alignment of nucleic acid molecules capable of binding to human CXCL8, including the K _D value determined by a competitive inhibitory binding assay;

In fig. 2 shows truncated derivatives of the nucleic acid molecule 315-F8-001, including the K _D value, and the relative binding activity to human CXCL8, as determined by competitive inhibitory binding assay and/or by surface plasmon resonance measurement;

In fig. 3 shows a kinetic assessment performed using a competitive inhibitory binding assay for D-nucleic acid molecules 315-F8-001 and 315-F8-002 with D-CXCL8;

In fig. 4 shows the binding of nucleic acid molecule 315-F8-002 to CXCL8 at (A) 25°C and (B) 37°C, as determined by surface plasmon resonance measurements. The association constant ka and the dissociation constant kd were obtained;

In fig. 5 shows the binding of the nucleic acid molecule 315-F8-002-PEG to CXCL8 at (A) 25°C and (B) 37°C, as determined by surface plasmon resonance measurements. The association constant ka and the dissociation constant kd were obtained;

In fig. 6 shows the binding selectivity of 315-F8-002 to CXCL8 determined using competitive binding assays by surface plasmon resonance measurements;

In fig. 7 shows the inhibition of CXCL8-induced chemotaxis of CXCR2-expressing cells by the nucleic acid molecule 315-F8-002-PEG;

In fig. 8 is a bar graph illustrating the quantification of CXCL8 using nucleic acid molecule 315-F8-002 (FIG. 8A) and quantification of CCL5 using nucleic acid molecule 315-F8-002 (FIG. 8B); And

In fig. 9 is a bar graph illustrating the binding of L-aptamer 315-F8-002 (FIG. 9A) and aptamer 8A-35 (FIG. 9B) to soluble CXCL8 and CXCL1 at various concentrations.

Example 1 . Nucleic acids capable of binding to human CXCL8

Several CXCL8-binding nucleic acids and their derivatives have been identified, the nucleotide sequences of which are shown in FIG. 1-2. Nucleic acids binding to CXCL8 were tested as D-aptamers (D-nucleic acids) and L-aptamers (L-nucleic acids), resulting in the synthesis of D-nucleic acids and L-nucleic acids as described in Example 2.

Nucleic acids that bind to CXCL8 have been characterized:

a) as D-aptamers using inhibitory binding assays with biotinylated D-CXCL8 (SEQ ID NO: 1) as shown in Example 3; And

b) as L-aptamers by surface plasmon resonance (SPR) measurement (Example 4) and by in vitro assay using cells expressing the human CXCR2 receptor, both of which used L-CXCL8 (SEQ ID NO: 2) (Example 5).

The nucleic acids thus obtained have several different sequences, and such sequences can be systematized or grouped as a family of sequences.

To define nucleotide sequence motifs, the IUPAC abbreviations for ambiguous nucleotides are used:

S strong G or C W weak A or U(T) R purine G or A Y pyrimidine C or U (T) K keto G or U(T) M imino A or C B not A C or U (T) or G D not C A or G or U (T) H not G A or C or U (T) V not U A or C or G N All A or G or C or U (T)

Unless otherwise stated, any nucleic acid sequence or fragment sequence, respectively, is indicated in the 5'→3' direction.

As shown in Figures 1-2, CXCL8-binding nucleic acids contain the same central nucleotide fragment defining a potential CXCL8-binding motif, where Figure 1 shows various sequences from a family of sequences and Figure 2 shows truncated derivatives of the CXCL8-binding nucleic acid acids 315-F8-001, including CXCL8-binding nucleic acid 315-F8-002-PEG.

In general, nucleic acid molecules that bind to CXCL8 contain, at their 5' end and 3' end, terminal nucleotide fragments: a first terminal nucleotide fragment and a second terminal nucleotide fragment. The first terminal nucleotide fragment and the second terminal nucleotide fragment can hybridize with each other, and after such hybridization, a double-stranded structure is formed. However, such hybridization is not necessarily observed in the molecule in vivo and in vitro .

The three nucleotide fragments of the nucleic acid molecules that bind to CXCCL8, namely, the first terminal nucleotide fragment, the central nucleotide fragment and the second terminal nucleotide fragment are located with respect to each other in the 5' → 3' direction: the first terminal nucleotide fragment - the central nucleotide fragment - second terminal nucleotide fragment. However, alternatively, the first terminal nucleotide fragment, the central nucleotide fragment and the second terminal nucleotide fragment are located with respect to each other in a 5' → 3' direction: the second terminal nucleotide fragment - the central nucleotide fragment - the first terminal nucleotide fragment.

The sequences of certain fragments may differ between CXCL8-binding nucleic acid molecules, which affects the binding affinity for CXCL8. Based on the binding assay of various nucleic acid molecules binding to CXCL8, it was found that the central nucleotide fragment and their nucleotide sequences described below, individually, and more preferably along their entire length, play an important role in binding to CXCL8.

The nucleic acids that bind to CXCL8 are composed of ribonucleotides (as shown in Figures 1-2).

As shown in Figure 1, the central nucleotide fragment sequences of the CXCCL8-binding nucleic acids 315-H9-001, 315-F11-001 and 315-F8-001 are slightly different from each other:

5' GGAAGUACGUGGAAAGCCAAUGAGUGUGUCCCG 3' (315-H9-001),

5' GGAAGUACGUGGAAAGCCGAUGAGUGUGUCCCG 3' (315-F11-001),

5' GGAAGUACGUGGAAAGCCGAAAGUGUGUCCCG 3' (315-F8-001).

The central nucleotide fragments of the CXCL8-binding nucleic acids of the invention contain 32-33 nucleotides and can be arranged in the consensus sequence: 5' GGAAGUACGUGGAAAGCCRA(X _U )RAGUGUGUCCCG 3', where X _U is U or absent.

Nucleic acids that bind to CXCL8 with the best binding affinity to CXCL8, that is, CXCL8-binding nucleic acids with the highest binding affinity to CXCL8 or the lowest dissociation constant Kd for CXCL8, contain central nucleotide fragments with the sequences 5' GGAAGUACGUGGAAAGCCGAAAGUGUGUCCCG 3' ( 315-F8-001) and 5' GGAAGUACGUGGAAAGCCAAUGAGUGUGUCCCG 3' (315-H9-001), where the central nucleotide fragments with the sequence 5' GGAAGUACGUGGAAAGCCGAAAGUGUGUCCCG 3' (315-F8-001) confer the best binding affinity of the CXCL8-binding nucleic acid to CXCL8.

As shown in Figures 1 and 2, the first terminal nucleotide fragment and the second terminal nucleotide fragment of the CXCL8-binding nucleic acids contain six nucleotides, five nucleotides, four nucleotides or three nucleotides, whereby these fragments do not necessarily hybridize with each other, and after such hybridization, a double-stranded structure is formed. This double-stranded structure may consist of six, five, four or three base pairs. However, such hybridization is not necessarily observed in a given molecule.

As shown in Figure 1, the first and second nucleotide fragments of CXCL8-binding nucleic acids 315-H9-001, 315-F11-001 AND 315-F8-001 contain six nucleotides, respectively:

a) CXCL8-binding nucleic acids 315-H9-001 and 315-F8-001 - the first terminal nucleotide fragment contains the nucleotide sequence 5'GCUGAC 3', and the second terminal nucleotide fragment contains the nucleotide sequence 5'GUCAGC 3';

b) CXCL8-binding nucleic acid 315-F11-001 - the first terminal nucleotide fragment contains the nucleotide sequence 5'GCUGAC 3', and the second terminal nucleotide fragment contains the nucleotide sequence 5'GUGAGC 3'.

As shown in Figure 2, the first and second nucleotide fragments of the CXCL8-binding nucleic acids contain five, four or three nucleotides, respectively:

a) CXCL8-binding nucleic acid 315-F8-002 - the first terminal nucleotide fragment contains the nucleotide sequence 5'CUGAC 3', and the second terminal nucleotide fragment contains the nucleotide sequence 5'GUCAG 3';

b) CXCL8-binding nucleic acid 315-F8-005 - the first terminal nucleotide fragment contains the nucleotide sequence 5'GUGAC 3', and the second terminal nucleotide fragment contains the nucleotide sequence 5'GUCAC 3';

c) CXCL8-binding nucleic acid 315-F8-003 - the first terminal nucleotide fragment contains the nucleotide sequence 5'-UGAC-3', and the second terminal nucleotide fragment contains the nucleotide sequence 5'GUCA 3';

d) CXCL8-binding nucleic acid 315-F8-006 - the first terminal nucleotide fragment contains the nucleotide sequence 5'GGAC 3', and the second terminal nucleotide fragment contains the nucleotide sequence 5'GUCC 3';

e) CXCL8-binding nucleic acid 315-F8-007 - the first terminal nucleotide fragment contains the nucleotide sequence 5'GCAC 3', and the second terminal nucleotide fragment contains the nucleotide sequence 5'GUGC 3';

f) CXCL8-binding nucleic acid 315-F8-008 - the first terminal nucleotide fragment contains the nucleotide sequence 5'GGUC 3', and the second terminal nucleotide fragment contains the nucleotide sequence 5'GACC 3';

g) CXCL8-binding nucleic acid 315-F8-009 - the first terminal nucleotide fragment contains the nucleotide sequence 5'UTGGC 3', and the second terminal nucleotide fragment contains the nucleotide sequence 5'GCCA 3';

h) CXCL8-binding nucleic acid 315-F8-004 - the first terminal nucleotide fragment contains the nucleotide sequence 5'-GAC-3', and the second terminal nucleotide fragment contains the nucleotide sequence 5'-GUC-3'.

The first terminal nucleotide fragment and the second terminal nucleotide fragment of all tested CXCL8-binding nucleic acid molecules can be systematized in the following general formula for the first terminal nucleotide fragment and the second terminal nucleotide fragment:

the general formula for the first terminal nucleotide fragment is 5'-Z ₁ Z ₂ Z ₃ Z ₄ Z ₅ C-3', and the general formula for the second terminal nucleotide fragment is 5'-GZ ₆ Z ₇ Z ₈ Z ₉ Z ₁₀ -3', where Z ₁ is G or absent, Z ₂ is S or absent; Z ₃ is K or absent, Z ₄ is S; Z ₅ is D; Z ₆ represents H; Z ₇ represents S; Z ₈ is M or absent; Z ₉ is S or absent, and Z ₁₀ is C or absent;

where in the first preferred embodiment of the invention:

q) Z ₁ is G, Z ₂ is S, Z ₃ is K, Z ₄ is S, Z ₅ is D, Z ₆ is H, Z ₇ is S, Z ₈ is M, Z ₉ represents S, and Z ₁₀ represents C; or

r) Z ₁ is absent, Z ₂ is S, Z ₃ is K, Z ₄ is S, Z ₅ is D, Z ₆ is H, Z ₇ is S, Z ₈ is M, Z ₉ is S and Z ₁₀ is missing; or

s) Z ₁ is absent, Z ₂ is absent, Z ₃ is K, Z ₄ is S, Z ₅ is D, Z ₆ is H, Z ₇ is S, Z ₈ is M, Z ₉ is absent, and Z ₁₀ is missing; or

t) Z ₁ is missing, Z ₂ is missing, Z ₃ is missing, Z ₄ is S, Z ₅ is D, Z ₆ is H, Z ₇ is S, Z ₈ is missing, Z ₉ is missing, and Z ₁₀ is missing ; or

u) Z ₁ is absent, Z ₂ is S, Z ₃ is K, Z ₄ is S, Z ₅ is D, Z ₆ is H, Z ₇ is S, Z ₈ is M, Z ₉ represents S, and Z ₁₀ represents C; or

v) Z ₁ is absent, Z ₂ is absent, Z ₃ is K, Z ₄ is S, Z ₅ is D, Z ₆ is H, Z ₇ is S, Z ₈ is M, Z ₉ is S, and Z ₁₀ represents C; or

w) Z ₁ is missing, Z ₂ is missing, Z ₃ is missing, Z ₄ is S, Z ₅ is D, Z ₆ is H, Z ₇ is S, Z ₈ is M, Z ₉ is S, and Z ₁₀ represents C; or

x) Z ₁ is G, Z ₂ is S, Z ₃ is K, Z ₄ is S, Z ₅ is D, Z ₆ is H, Z ₇ is S, Z ₈ is M, Z ₉ is S and Z ₁₀ is absent; or

y) Z ₁ is G, Z ₂ is S, Z ₃ is K, Z ₄ is S, Z ₅ is D, Z ₆ is H, Z ₇ is S, Z ₈ is M, Z ₉ is missing and Z ₁₀ is missing; or

z) Z ₁ is G, Z ₂ is S, Z ₃ is K, Z ₄ is S, Z ₅ is D, Z ₆ is H, Z ₇ is S, Z ₈ is absent, Z ₉ missing and Z ₁₀ missing; or

aa) Z ₁ is absent, Z ₂ is S, Z ₃ is K, Z ₄ is S, Z ₅ is D, Z ₆ is H, Z ₇ is S, Z ₈ is M, Z ₉ missing and Z ₁₀ missing; or

bb) Z ₁ is absent, Z ₂ is S, Z ₃ is K, Z ₄ is S, Z ₅ is D, Z ₆ is H, Z ₇ is S, Z ₈ is absent, Z ₉ is absent, and Z ₁₀ is missing; or

cc) Z ₁ is absent, Z ₂ is absent, Z ₃ is K, Z ₄ is S, Z ₅ is D, Z ₆ is H, Z ₇ is S, Z ₈ is M, Z ₉ is S, and Z ₁₀ is missing; or

dd) Z ₁ is missing, Z ₂ is missing, Z ₃ is missing, Z ₄ is S, Z ₅ is D, Z ₆ is H, Z ₇ is S, Z ₈ is M, Z ₉ is S, and Z ₁₀ is missing; or

ee) Z ₁ is absent, Z ₂ is absent, Z ₃ is K, Z ₄ is S, Z ₅ is D, Z ₆ is H, Z ₇ is S, Z ₈ is absent, Z ₉ is absent, and Z ₁₀ missing; or

ff) Z ₁ is missing, Z ₂ is missing, Z ₃ is missing, Z ₄ is S, Z ₅ is D, Z ₆ is H, Z ₇ is S, Z ₈ is M, Z ₉ is missing, and Z ₁₀ is missing.

Nucleic acids that bind to CXCL8 with the best binding affinity for CXCL8, that is, CXCL8-binding nucleic acids with the highest binding affinity for CXCL8 or the lowest dissociation constant Kd for CXCL8, include the following first and second terminal nucleotide fragments:

a) CXCL8-binding nucleic acids 315-H9-001 and 315-F8-001 - the first terminal nucleotide fragment contains the nucleotide sequence 5'GCUGAC-3', and the second terminal nucleotide fragment contains the nucleotide sequence 5'GUCAGC-3';

b) CXCL8-binding nucleic acid 315-F8-002 - the first terminal nucleotide fragment contains the nucleotide sequence 5'CUGAC 3', and the second terminal nucleotide fragment contains the nucleotide sequence 5'GUCAG 3';

c) CXCL8-binding nucleic acid 315-F8-005 - the first terminal nucleotide fragment contains the nucleotide sequence 5'GUGAC 3', and the second terminal nucleotide fragment contains the nucleotide sequence 5'GUCAC 3'.

Kd values for CXCL8-binding nucleic acids 315-F8-001, 315-H9-001 and 315-F11-001 (containing six-nucleotide terminal fragments) were measured in the range of 1.5-12.5 nM, as determined by the assay for competitive inhibitory binding (Figures 1-3). In a direct inhibition assay, CXCL8-binding nucleic acid 315-F8-001 had a Kd value of 0.8 nM. When measured by surface plasmon resonance, which is a more sensitive method for determining Kd values, the Kd value for CXCL8-binding nucleic acid 315-F8-001 was 0.22 nM at 25°C and 0.68 nM at 37°C (Figure 2 ).

Surprisingly, it was found that truncation of the first terminal fragment from six to five nucleotides and truncation of the second terminal fragment from six to five nucleotides did not negatively affect the binding affinity for CXCL8. Binding affinity of CXCL8-binding nucleic acid 315-F8-002 measured by competitive inhibition assay (Kd 1.1 nM; Figures 2 and 3) and by surface plasmon resonance measurement (Kd 0.22 nM at 25°C and 0.75 nM at 37°C; Figures 2 and 4), is within the binding affinities determined for CXCL8-binding nucleic acid 315-F8-001 (see Kd values for 315-F8-001 above).

Attempts to further truncation and/or mutation of the terminal fragments of the CXCL8-binding nucleic acid 315-F8-002 resulted in the production of CXCL8-binding nucleic acids 315-F8-003, 315-F8-004, 315-F8-005, 315-F8-006 , 315-F8-007, 315-F8-008 and 315-F8-009 with lower (315-F8-003, 315-F8-004, 315-F8-006, 315-F8-007, 315-F8- 008 and 315-F8-009, 315-F8-007, 315-F8-008 and 315-F8-009) or similar (315-F8-005) binding affinities to CXCL8 (Figure 2).

For the 40 kDa 5'-PEGized variant of CXCL8-binding nucleic acid 315-F8-002, namely CXCL8-binding nucleic acid 315-F8-002-PEG (also called AON-S08), Kd is 0.2 nM at 25 °C and 0.8 nM at 37°C was determined by surface plasmon resonance measurements (Figures 2 and 5).

CXCL8 is one of several ELR-positive human CXC chemokines. In addition to CXCL8, the ELR-positive human CXC chemokines: CXCL1, CXCL2, CXCL3, CXCL5, CXCL6, CXCL7 bind to the CXCL8 receptor CXCR2, where the ELR-positive human CXC chemokines: CXCL6 and CXCL7 also bind to the CXCL8 receptor CXCR1. To confirm the specificity and selectivity of the binding properties of CXCL8-binding nucleic acid 315-F8-002, the binding affinity of CXCL8-binding nucleic acid 315-F8-002 to ELR-positive human CXC chemokines: CXCL1, CXCL2, CXCL3, CXCL5, CXCL6, CXCL7 was tested using SPR in a competitive binding assay (Example 4). CXCL8-binding nucleic acid 315-F8-002 did not bind to any of the other ELR-positive human CXC chemokines: CXCL1, CXCL2, CXCL3, CXCL5, CXCL6, CXCL7 (Figure 6).

CXCL8-binding nucleic acid 315-F8-002-PEG inhibited CXCL8-induced chemotaxis of CXCR2-expressing BA/F3 cells with an average inhibition constant IC ₅₀ of 0.26 nM (Figure 7, Example 5).

Biosensor immobilized with CXCL8-binding nucleic acid 315-F8-002 was used for detection and quantification of CXCL8. The limit of detection (mean baseline value + 3× standard deviation from baseline values) was <98 pM for CXCL8, and the lower limit of quantification (mean baseline value + 10× standard deviation from baseline values) was also <98 pM for CXCL8 ( Figure 8A ,Example 6). To demonstrate the specificity of the immobilized CXCL8-binding nucleic acid 315-F8-002, another chemokine (CCL5) was used as an analyte. CCL5 binding was not measured at concentrations up to 12.5 nM (Figure 8B, Example 6).

Example 2 : Synthesis of D- and L-aptamers

Solid-phase synthesis on a laboratory scale

D-aptamers (D-nucleic acids) and L-aptamers (L-nucleic acids) were obtained by solid-phase synthesis using an ABI 394 synthesizer (Applied Biosystems, Foster City, CA, USA) by performing a chemical addition reaction of 2'TBDMS-RNA- phosphoramidite (Damha and Ogilvie, 1993). L-rA(N-Bz)-, L-rC(Ac)-, L-rG(N-ibu)-, L-rU-, D-rA(N-Bz)-, D-rC(Ac)- , D-rG(N-ibu)-, D-rU-phosphoramidites were purchased from ChemGenes, Wilmington, MA. D- and L-aptamers were purified using gel electrophoresis.

Large-scale solid-phase synthesis

D-aptamers were prepared by solid-phase synthesis using an AktaPilotlOO synthesizer (Amersham Biosciences; General Electric Healthcare, Freiburg) by coupling 2′TBDMS-RNA and DNA phosphoramidite (Damha and Ogilvie, 1993). L-rA(N-Bz)-, L-rC(Ac)-, L-rG(N-ibu), and L-rU- were purchased from ChemGenes, Wilmington, MA. The 5'-amino modifier was purchased from American International Chemicals Inc (Framingham, MA, USA). Synthesis of unmodified or 5′-amino-modified L-aptamer was started from L-riboA, L-riboC, L-riboG, or L-riboU modified with 1000A pore size CPG (Link Technology, Glasgow, UK). To bind RNA phosphoramidites (15 min per cycle), 0.3 M benzylthiotetrazole (CMS-Chemicals, Abingdon, UK) in acetonitrile and 2 equivalents of a corresponding 0.2 M solution of phosphoramidite in acetonitrile were used. A cycle of oxidation-capping reactions was carried out. The following standard solvents and reagents purchased from Biosolve (Valkenswaard, NL) were used to synthesize oligonucleotides. The L-aptamer was synthesized DMT-ON; and after deprotection, it was purified by preparative RP-HPLC (Wincott et al. 1995) using Sourcel5RPC medium (Amersham). The 5'-DMT group was removed with 80% acetic acid (30 minutes at room temperature). In the case of 5'-amino-modified L-aptamers, the 5'-MMT group was removed with 80% acetic acid (90 minutes at room temperature). Aqueous 2M NaOAc was then added and the L-aptamer was desalted by tangential flow filtration using a 5K regenerated cellulose membrane (Millipore, Bedford, MA).

PEGylation of L-aptamers

To increase the residence time of the L-aptamer in plasma in vivo , the L-aptamer was covalently linked to a 40 kDa polyethylene glycol moiety (PEG) at the 5' end. For PEGylation (technical details of the PEGylation method can be found in European patent application EP1306382), the purified 5'-amino-modified L-aptamer was dissolved in a mixture of H ₂ O (2.5 ml), DMF (5 ml) and buffer A (5 ml , obtained by mixing citric acid × H ₂ O [7 g], boric acid [3.54 g], phosphoric acid [2.26 ml] and 1 M NaOH [343 ml] and adding water to a final volume of 1 L; pH =8.4 was adjusted by adding 1 M HCl).

The pH of the L-aptamer solution was adjusted to 8.4 using 1 M NaOH. 40 kDa PEG-NHS ester (Jenkem Technology, Allen, TX, USA) was then added at 37°C every 30 minutes in six parts of 0.25 equivalents until a maximum yield of 75-85% was achieved. The pH of the reaction mixture was maintained at 8-8.5 with 1M NaOH during the addition of the PEG-NHS ester.

The reaction mixture was mixed with 4 ml of urea solution (8 M) and 4 ml of buffer B (0.1 M triethylammonium acetate in H ₂ O) and heated to 95°C for 15 minutes. The PEGylated L-aptamer was then purified by RP-HPLC in Source 15 RPC (Amersham) acetonitrile gradient (buffer B; buffer C: 0.1 M triethylammonium acetate in acetonitrile). Excess PEG was eluted at 5% Buffer C and the PEGylated L-aptamer was eluted at 10-15% Buffer C. Product fractions >95% pure (as determined by HPLC) were pooled and mixed with 40 ml 3M NaOAc. The PEGylated L-aptamer was desalted by tangential flow filtration (using a 5K regenerated cellulose membrane, Millipore, Bedford, MA).

Example 3 : Inhibitory binding assays

Direct inhibition assay to determine binding constants of D-aptamers

Direct inhibition assays (SEQ ID NO: 1) were performed to determine the binding affinity of D-aptamers to D-CXCL8(C-bio) (SEQ ID NO: 1). For this purpose, D-aptamers were radiolabeled with T4 polynucleotide kinase (Invitrogen, Karlsruhe, Germany) using [γ- ^32P ]-ATP (Hartmann Analytic, Braunschweig, Germany). The specific radioactivity of labeled D-aptamers was 110,000–300,000 cpm/pmol. Labeled D-aptamers were incubated at a concentration of 0.2 nM in selection buffer (20 mM Tris, pH 7.4, 150 mM NaCl, 5 mM KCl, 1 mM _MgCl2 , 1 mM _CaCl2 , 0.1% Tween 20, 100 µg/ml essential fatty acids without bovine serum albumin, 200 mg/ml yeast RNA) along with varying amounts of D-CXCL8(C-bio) ranging from 0.0192 to 300 nM for 1.5-2 hours at 37 °C. D-CXCL8(C-bio)/D-aptamer complexes were immobilized on NAag ⁺ beads (neutravidin plus agarose from Pierce Biotechnology, Rockford, USA) pre-equilibrated in selection buffer. After removal of the supernatant and appropriate washing, the radioactivity associated with the spheres was measured in a scintillation counter (LS6500; Beckman Coulter, Fullton, USA). The amount of immobilized labeled D-aptamer (% of initial amount) was plotted against the concentration of D-CXCL8(C-Bio), and dissociation constants Kd were obtained using software algorithms (GRAFIT; Erithacus Software; Surrey UK) with an assumed stoichiometry of 1: 1.

For CXCL8-binding aptamers 315-H9-001 AND 315-F8-001, Kd values of 1.7 nM and 0.8 nM were determined by direct inhibition assay, respectively.

Competitive inhibition assay for ranking and determining binding constants of D-aptamers .

Competitive inhibition assays were performed to compare the affinities of different D-CXCL8(C-bio)-binding D-aptamers. For this purpose, the reference D-aptamer was radiolabeled (as described above) and incubated at 37°C with D-CXCL8(C-bio) in selection buffer under conditions that promote the transmission of an unsaturated binding signal after immobilization on NAag ⁺ and washing (e.g. 3 nM D-CXCL8(C-Bio), 0.15 nM labeled D-aptamer). The addition of excess unlabeled D-aptamer, which competes with the labeled D-aptamer for binding to D-CXCL8(C-Bio), results in a decrease in the amount of radioactivity associated with the spheres after immobilization and washing. The degree of signal reduction depends on the amount and affinity of the unlabeled D-aptamer. Competitive inhibition assays were performed for ranking experiments and to determine dissociation constants (Kd) of selected D-aptamers. Dissociation constants Kd were determined by plotting the fraction (%) of labeled D-aptamer bound to D-CXCL8(C-bio) versus the concentration of an unlabeled competing D-aptamer. Data analyzes were performed using GRAFIT.

The results of inhibitory binding assays are presented in Example 1 and shown in FIG. 1, 2 and 3, where the CXCL8-binding nucleic acid 315-F8-001 was used as the labeled D-aptamer.

Example 4 : Surface Plasmon Resonance (SPR) Binding Assays

Surface plasmon resonance measurements were performed on a Biacore 2000 instrument (GE Healthcare) set at a constant temperature of 25°C or 37°C. Proteins were immobilized on CM4 sensor chips (GE Healthcare) via an amine addition reaction. The surface of the sensor chip was activated by injecting a 1:1 mixture of 0.4 M EDC (1-ethyl-3-(3-dimethylaminopropyl)carbodiimide; GE) and 0.1 M NHS (N-hydroxysuccinimide; GE). The maximum observed response for a covalently immobilized peptide or protein was approximately 500 RU. Flow cells were blocked with 1 M ethanolamine hydrochloride (GE, BR-1000-50). Non-covalently bound peptide or protein was also removed using this procedure. A flow cell with a surface not treated with dextran and a flow cell with a surface blocked with ethanolamine were used as controls. Before measurement, the sensor chip was treated twice with degassed physiological working buffer (20 mM Tris, pH 7.4, 150 mM NaCl, 5 mM KCl, 1 mM MgCl ₂ and 1 mM CaCl ₂ ) and performed at least three injection and regeneration cycles ( 5M NaCl).

The binding affinities of L-aptamers were measured by injection of a series of concentrations of 1000, 500, 250, 125, 62.5, 31.3, 15.6, 7.8, 3.9, 1.95, 0.98, 0.49, 0.24, 0.12, 0.06 nM and registration of the association and dissociation phases of binding events. The resulting binding curves were plotted according to the Langmuir 1:1 stoichiometric binding model to determine the binding kinetics constants (association constants ka; dissociation constants kd), which are used to calculate the dissociation constant (Kd=kd/ka). Data analysis was performed using the computer program BIAevaleation 3.1.1 (BIACORE AB, Uppsala, Sweden) with a constant refractive index (RI) and an initial mass transfer coefficient Kt of 1 × 10 ⁷ (RU × M ^-1 s ^-1 ).

The binding selectivity of the L-aptamer was assessed using competitive binding assays. For this purpose, human chemokines CXCL1, CXCL2, CXCL3, CXCL5, CXCL6, CXCL7, and CXCL8 (purchased from RnD Systems, PeproTech, or ProSpecTech) were separately injected at 40 nM along with the CXCL8-binding nucleic acid L-aptamer 315-F8-002 ( 0.25 nM) to evaluate the competitive binding of the L-aptamer to immobilized CXCL8. As a control, chemokines were injected in the absence of L-aptamer to monitor binding to the dextran sensor matrix and/or immobilized CXCL8. Binding of 315-F8-002 to immobilized CXCL8 was detected at a predetermined time point 240 seconds after the end of the injection. Selected chemokines were immobilized, and the binding affinities of the CXCCL8-binding nucleic acid L-aptamer 315-F8-002 were determined as described above.

The results of the SPR binding assays are reported in Example 1 and shown in Figures 2, 4, 5 and 6.

Example 5 : Pharmacology in vitro - Chemotaxis analysis

The mouse pro-B cell line BA/F3 was stably transfected with a plasmid encoding human CXCR2. For chemotaxis assays, recombinant CXCL8 (0.5 nM) was preincubated with the CXCL8-binding nucleic acid L-aptamer 315-F8-002-PEG at the indicated concentrations in HBH buffer (Hank's balanced salt solution (HBSS) + 1 mg/ml BSA + 20 mM HEPES) in the lower compartments of a 96-well Corning Transwell plate with 5 μm pores (Costar Corning, NY) at 37°C for 20-30 minutes. CXCR2 ⁺ cells in HBH buffer were added to the upper compartments, and these cells were able to migrate at 37°C for 3 hours. After removal of the upper compartments, 50 μM resasurin (Sigma-Aldrich) in PBS was added to the lower compartments and incubated at 37°C for 2.5 hours. Fluorescence was measured at 590 nm (excitation wavelength 544 nm). Background-corrected and normalized fluorescence values were plotted against L-aptamer concentration. _IC50 inhibition constants (L-aptamer concentrations required for half-maximal inhibition) were determined by nonlinear regression (4-parameter fit) using Prism 5 (GraphPad Software, San Diego, CA).

The results of the chemotaxis assays are given in Example 1 and shown in Figure 7.

Example 6 : Biosensor CXCL8

SPR sensor chips coated with an azide-terminated carbon nanomembrane were prepared, and the CXCL8-binding L-aptamer 315-F8-002-DBCO (DBCO-modified version of 315-F8-002-amino) was immobilized from a 20 μM solution in 2 M NaCl on a flow cell of a commercially available Biacore system (up to approximately 1000 RU). As a reference, a nonfunctional L-aptamer (DBCO-modified reverse 315-F8-002-amino) was immobilized to a similar amount on another flow cell. Surfaces were passaged by immobilizing DBCO-modified methoxy-terminated straight chain polyethylene glycol (Molecular weight: 5 kDa). Injection of a series of concentrations of the analyte CXCL8, which was injected into a standard sample buffer (Copan Universal Transport Medium) supplemented with 0.1% (w/v) Tween 20, produced a dose-dependent increase in signal. The temperature during the measurement was 25°C, the flow rate was 30 μl/min, and the working buffer was the measurement buffer (20 mM Tris, pH 7.4, 150 mM NaCl, 5 mM KCl, 1 mM MgCl ₂ 1 mM CaCl ₂ , 0.1% (w/v) Tween 20). The analyte was left to associate for 10 minutes and then for 2.5 minutes. for dissociation in measurement buffer. The signal at the end of the dissociation phase was calculated by subtracting the signal obtained from the reference flow cell.

Results for the CXCL8 biosensor are reported in Example 1 and shown in Figure 8.

Example 7 : SPR binding assay to compare binding to soluble chemokines

The inhibitory activity of an aptamer depends on its ability to bind to a target in solution. Here, a competitive binding SPR assay was performed to determine the binding of the L-aptamer CXCL8-binding nucleic acid 315-F8-002 according to the invention and the binding of the previously published aptamer 8A-35 (Sung, Biomaterials 2014) to soluble CXCL8 and CXCL1. For this purpose, SPR measurements were performed as described in Example 4 using CXCL8 immobilized on a Cl-sensor chip (GE Healthcare) by an amine addition reaction. Binding of 315-F8-002 (3 nM) or 8A-35 (1.6 nM) to immobilized CXCL8 resulted in a binding reaction of approximately 20 response units. In 8A-35 binding assays, sensor chips were regenerated using 1M CaCl ₂ solution. To assess the binding of both aptamers to their target in solution, soluble CXCL8 was simultaneously injected at an equimolar concentration (1:1 ratio) or in 5-fold excess (1:5 ratio) to assay for competitive binding of the aptamer to immobilized CXCL8. Soluble CXCL1 binding was used as a control. All measurements were carried out at 37°C. Responses to binding to immobilized CXCL8 were recorded after 240 sec. after injection.

Binding of 315-F8-002 to immobilized CXCL8 was completely blocked (>90%) by an equimolar concentration of soluble CXCL8 (Figure 9A). Conversely, 8A-35 binding was only partially blocked by soluble CXCL8 at equimolar concentration (ca. 30%) and 5-fold excess (ca. 45%) (Figure 9B). This indicates that 315-F8-002 has higher binding affinity for soluble CXCL8 compared to 8A-35. Reduced binding affinity to soluble CXCL8 (compared to surface immobilized CXCL8) may explain the poor inhibitory activity of 8A-358 observed in the CXCL8-induced neutrophil migration assay (Sung, Biomaterials 2014).

Soluble CXCL1 was used as a control because 315-F8-002 was shown not to be cross-reactive with other ELR-positive chemokines (Figure 6). Consistent with this, 315-F8-002 binding was not blocked by soluble CXCL1 (Figure 9A). Surprisingly, it was found that 8A-358 binding was blocked by soluble CXCL1 with the same efficiency as soluble CXCL8 (ca. 25% at equimolar concentration and ca. 35% at 5-fold excess concentration (Figure 9B). This indicates that that 8A-35, unlike 315-F8-002, does not selectively bind to CXCL8 and will not further inhibit CXCL8 in solution.

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The project underlying the development of this application was the result of a study carried out by Horizon of the European Union in 2020, in accordance with the innovation program implemented under grant No. 634415.

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LIST OF SEQUENCES

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<400> 23

ggucggaagu acguggaaag ccgaaagugu gucccggacc

40

<210> 24

<211> 40

<212> RNA

<213> Artificial sequence

<220>

<223> Synthetic

<400> 24

uggcggaagu acguggaaag ccgaaagugu gucccggcca

40

<210> 25

<211> 42

<212> RNA

<213> Artificial sequence

<220>

<223> Synthetic

<220>

<221> misc_feature

<223> PEG linked via an aminohexyl linker to

5ʹ-end

<400> 25

cugacggaag uacguggaaa gccgaaagug ugucccgguc ag

42

<210> 26

<211> 42

<212> RNA

<213> Artificial sequence

<220>

<223> Synthetic

<220>

<221> misc_feature

<223> aminohexyl linker attached to the 5' end

<400> 26

cugacggaag uacguggaaa gccgaaagug ugucccgguc ag

42

<210> 27

<211> 33

<212> RNA

<213> Artificial sequence

<220>

<223> Synthetic

<220>

<221> misc_feature

<222> (21)..(21)

<223> U is U or missing

<400> 27

ggaaguacgu ggaaagccra uraguguc ccg

33

<210> 28

<211> 33

<212> RNA

<213> Artificial sequence

<220>

<223> Synthetic

<400> 28

ggaaguacgu ggaaagccaa ugaguguguc ccg

33

<210> 29

<211> 33

<212> RNA

<213> Artificial sequence

<220>

<223> Synthetic

<400> 29

ggaaguacgu ggaaagccga ugaguguguc ccg

33

<210> 30

<211> 32

<212> RNA

<213> Artificial sequence

<220>

<223> Synthetic

<400> 30

ggaaguacgu ggaaagccga aagugugucc cg

32

<210> 31

<211> 42

<212> RNA

<213> Artificial sequence

<220>

<223> Synthetic

<400> 31

gacuggcccu gugugaaagc cgaaaggugc augaaggcag uc

42

<210> 32

<211> 73

<212>PRT

<213> Homo sapiens

<220>

<221> MISC_FEATURE

<222> (1)..(73)

<223> CXCL1

<400> 32

Ala Ser Val Ala Thr Glu Leu Arg Cys Gln Cys Leu Gln Thr Leu Gln

1 5 10 15

Gly Ile His Pro Lys Asn Ile Gln Ser Val Asn Val Lys Ser Pro Gly

20 25 30

Pro His Cys Ala Gln Thr Glu Val Ile Ala Thr Leu Lys Asn Gly Arg

35 40 45

Lys Ala Cys Leu Asn Pro Ala Ser Pro Ile Val Lys Lys Ile Ile Glu

50 55 60

Lys Met Leu Asn Ser Asp Lys Ser Asn

65 70

<210> 33

<211> 73

<212>PRT

<213> Homo sapiens

<220>

<221> MISC_FEATURE

<222> (1)..(73)

<223> CXCL2

<400> 33

Ala Pro Leu Ala Thr Glu Leu Arg Cys Gln Cys Leu Gln Thr Leu Gln

1 5 10 15

Gly Ile His Leu Lys Asn Ile Gln Ser Val Lys Val Lys Ser Pro Gly

20 25 30

Pro His Cys Ala Gln Thr Glu Val Ile Ala Thr Leu Lys Asn Gly Gln

35 40 45

Lys Ala Cys Leu Asn Pro Ala Ser Pro Met Val Lys Lys Ile Ile Glu

50 55 60

Lys Met Leu Lys Asn Gly Lys Ser Asn

65 70

<210> 34

<211> 73

<212>PRT

<213> Homo sapiens

<220>

<221> MISC_FEATURE

<222> (1)..(73)

<223> CXCL3

<400> 34

Ala Ser Val Val Thr Glu Leu Arg Cys Gln Cys Leu Gln Thr Leu Gln

1 5 10 15

Gly Ile His Leu Lys Asn Ile Gln Ser Val Asn Val Arg Ser Pro Gly

20 25 30

Pro His Cys Ala Gln Thr Glu Val Ile Ala Thr Leu Lys Asn Gly Lys

35 40 45

Lys Ala Cys Leu Asn Pro Ala Ser Pro Met Val Gln Lys Ile Ile Glu

50 55 60

Lys Ile Leu Asn Lys Gly Ser Thr Asn

65 70

<210> 35

<211> 78

<212>PRT

<213> Homo sapiens

<220>

<221> MISC_FEATURE

<222> (1)..(78)

<223>CXCL5

<400> 35

Ala Gly Pro Ala Ala Ala Val Leu Arg Glu Leu Arg Cys Val Cys Leu

1 5 10 15

Gln Thr Thr Gln Gly Val His Pro Lys Met Ile Ser Asn Leu Gln Val

20 25 30

Phe Ala Ile Gly Pro Gln Cys Ser Lys Val Glu Val Val Ala Ser Leu

35 40 45

Lys Asn Gly Lys Glu Ile Cys Leu Asp Pro Glu Ala Pro Phe Leu Lys

50 55 60

Lys Val Ile Gln Lys Ile Leu Asp Gly Gly Asn Lys Glu Asn

65 70 75

<210> 36

<211> 77

<212>PRT

<213> Homo sapiens

<220>

<221> MISC_FEATURE

<222> (1)..(77)

<223> CXCL6

<400> 36

Gly Pro Val Ser Ala Val Leu Thr Glu Leu Arg Cys Thr Cys Leu Arg

1 5 10 15

Val Thr Leu Arg Val Asn Pro Lys Thr Ile Gly Lys Leu Gln Val Phe

20 25 30

Pro Ala Gly Pro Gln Cys Ser Lys Val Glu Val Val Ala Ser Leu Lys

35 40 45

Asn Gly Lys Gln Val Cys Leu Asp Pro Glu Ala Pro Phe Leu Lys Lys

50 55 60

Val Ile Gln Lys Ile Leu Asp Ser Gly Asn Lys Lys Asn

65 70 75

<210> 37

<211> 94

<212>PRT

<213> Homo sapiens

<220>

<221> MISC_FEATURE

<222> (1)..(94)

<223> CXCL7

<400> 37

Ser Ser Thr Lys Gly Gln Thr Lys Arg Asn Leu Ala Lys Gly Lys Glu

1 5 10 15

Glu Ser Leu Asp Ser Asp Leu Tyr Ala Glu Leu Arg Cys Met Cys Ile

20 25 30

Lys Thr Thr Ser Gly Ile His Pro Lys Asn Ile Gln Ser Leu Glu Val

35 40 45

Ile Gly Lys Gly Thr His Cys Asn Gln Val Glu Val Ile Ala Thr Leu

50 55 60

Lys Asp Gly Arg Lys Ile Cys Leu Asp Pro Asp Ala Pro Arg Ile Lys

65 70 75 80

Lys Ile Val Gln Lys Lys Leu Ala Gly Asp Glu Ser Ala Asp

85 90

<210> 38

<211> 68

<212>PRT

<213> Homo sapiens

<220>

<221> MISC_FEATURE

<222> (1)..(68)

<223> CCL5

<400> 38

Ser Pro Tyr Ser Ser Asp Thr Thr Pro Cys Cys Phe Ala Tyr Ile Ala

1 5 10 15

Arg Pro Leu Pro Arg Ala His Ile Lys Glu Tyr Phe Tyr Thr Ser Gly

20 25 30

Lys Cys Ser Asn Pro Ala Val Val Phe Val Thr Arg Lys Asn Arg Gln

35 40 45

Val Cys Ala Asn Pro Glu Lys Lys Trp Val Arg Glu Tyr Ile Asn Ser

50 55 60

Leu Glu Met Ser

65

<210> 39

<211> 42

<212> RNA

<213> Artificial sequence

<220>

<223> Synthetic

<220>

<221> misc_feature

<222> (42)..(42)

<223> PEG attached via aminohexyl linker

<400> 39

cugacggaag uacguggaaa gccgaaagug ugucccgguc ag

42

<210> 40

<211> 42

<212> RNA

<213> Artificial sequence

<220>

<223> Synthetic

<220>

<221> misc_feature

<222> (42)..(42)

<223> Attached aminohexyl linker

<400> 40

cugacggaag uacguggaaa gccgaaagug ugucccgguc ag

42

<210> 41

<211> 42

<212> RNA

<213> Artificial sequence

<220>

<223> Synthetic

<220>

<221> misc_feature

<222> (1)..(1)

<223> PEG attached via aminohexyl linker

<400> 41

gugacggaag uacguggaaa gccgaaagug ugucccgguc ac

42

<210> 42

<211> 42

<212> RNA

<213> Artificial sequence

<220>

<223> Synthetic

<220>

<221> misc_feature

<222> (1)..(1)

<223> Attached aminohexyl linker

<400> 42

gugacggaag uacguggaaa gccgaaagug ugucccgguc ac

42

<210> 43

<211> 42

<212> RNA

<213> Artificial sequence

<220>

<223> Synthetic

<220>

<221> misc_feature

<222> (42)..(42)

<223> PEG attached via aminohexyl linker

<400> 43

gugacggaag uacguggaaa gccgaaagug ugucccgguc ac

42

<210> 44

<211> 42

<212> RNA

<213> Artificial sequence

<220>

<223> Synthetic

<220>

<221> misc_feature

<222> (42)..(42)

<223> Aminohexyl linker

<400> 44

gugacggaag uacguggaaa gccgaaagug ugucccgguc ac

42

<210> 45

<211> 35

<212> RNA

<213> Artificial sequence

<220>

<223> Synthetic

<220>

<221> misc_feature

<223> All C are 2'-fluoro-C

<220>

<221> misc_feature

<223> All Us are 2'-fluoro-U

<400> 45

gggggcuuau cauuccauuu aguguuauga uaacc

35

<---

Claims (161)

1. An L-nucleic acid molecule capable of binding to human CXCL8, wherein the L-nucleic acid molecule contains a central nucleotide fragment and optionally a modifying group, wherein the central nucleotide fragment contains the nucleotide sequence: 5'-GGAAGUACGUGGAAAGCCRA(X _U )RAGUGUGUCCCG-3' [SEQ ID NO: 27], where X _U is U or is absent. 2. An L-nucleic acid molecule according to claim 1, where the central nucleotide fragment contains a nucleotide sequence selected from the group: a) 5’ GGAAGUACGUGGAAAGCCAAUGAGUGUGUCCCG 3’ [SEQ ID NO: 28], b) 5' GGAAGUACGUGGAAAGCCGAUGAGUGUGUCCCG 3' [SEQ ID NO: 29] and c) 5' GGAAGUACGUGGAAAGCCGAAAGUGUGUCCCG 3' [SEQ ID NO: 30]. 3. L-nucleic acid molecule according to claim 2, where the central nucleotide fragment contains the nucleotide sequence: 5' GGAAGUACGUGGAAAGCCGAAAGUGUGUCCCG 3' [SEQ ID NO: 30] or 5' GGAAGUACGUGGAAAGCCAAUGAGUGUGUCCCG 3' [SEQ ID NO: 28], and preferably 5' GGAAGUACGUGGAAAGCCGAAAGUGUGUCCCG 3' [SEQ ID NO: 30]. 4. L-nucleic acid molecule according to any one of paragraphs. 1-3, where the L-nucleic acid molecule contains in the 5' → 3' direction a first terminal nucleotide fragment, a central nucleotide fragment and a second terminal nucleotide fragment, where the first terminal nucleotide fragment contains from three to six nucleotides and the second terminal nucleotide fragment contains from three to six nucleotides, where preferable the first terminal nucleotide fragment contains from five to six nucleotides and the second terminal nucleotide fragment contains from five to six nucleotides, where is more preferable the first terminal nucleotide fragment contains five nucleotides and the second terminal nucleotide fragment contains five nucleotides. 5. The L-nucleic acid molecule according to claim 4, where the first terminal nucleotide fragment contains the nucleotide sequence 5'-Z ₁ Z ₂ Z ₃ Z ₄ Z ₅ C-3', and the second terminal nucleotide fragment contains the nucleotide sequence 5'-GZ ₆ Z ₇ Z ₈ Z ₉ Z ₁₀ -3', where Z ₁ is G or absent, Z ₂ is S or absent; Z ₃ is K or absent, Z ₄ is S, Z ₅ is D; Z ₆ represents H; Z ₇ represents S; Z ₈ is M or absent; Z ₉ is S or absent, and Z ₁₀ is C or absent; where preferable a) Z ₁ represents G; Z ₂ represents S; Z ₃ represents K; Z ₄ represents S; Z ₅ represents D; Z ₆ represents H; Z ₇ represents S; Z ₈ represents M; Z ₉ represents S and Z ₁₀ represents C; or b) Z ₁ is missing; Z ₂ represents S; Z ₃ represents K; Z ₄ represents S; Z ₅ represents D; Z ₆ represents H; Z ₇ represents S; Z ₈ represents M; Z ₉ is S and Z ₁₀ is missing; or c) Z ₁ is missing; Z ₂ is missing; Z ₃ represents K; Z ₄ represents S; Z ₅ represents D; Z ₆ represents H; Z ₇ represents S; Z ₈ represents M; Z ₉ missing and Z ₁₀ missing; or d) Z ₁ is missing; Z ₂ is missing; Z ₃ is missing; Z ₄ represents S; Z ₅ represents D; Z ₆ represents H; Z ₇ represents S; Z ₈ missing; Z ₉ missing and Z ₁₀ missing; or e) Z ₁ is missing; Z ₂ represents S; Z ₃ represents K; Z ₄ represents S; Z ₅ represents D; Z ₆ represents H; Z ₇ represents S; Z ₈ represents M; Z ₉ represents S and Z ₁₀ represents C; or f) Z ₁ is missing; Z ₂ is missing; Z ₃ represents K; Z ₄ represents S; Z ₅ represents D; Z ₆ represents H; Z ₇ represents S; Z ₈ represents M; Z ₉ represents S and Z ₁₀ represents C; or g) Z ₁ is missing; Z ₂ is missing; Z ₃ is missing; Z ₄ represents S; Z ₅ represents D; Z ₆ represents H; Z ₇ represents S; Z ₈ represents M; Z ₉ represents S and Z ₁₀ represents C; or h) Z ₁ represents G; Z ₂ represents S; Z ₃ represents K; Z ₄ represents S; Z ₅ represents D; Z ₆ represents H; Z ₇ represents S; Z ₈ represents M; Z ₉ represents S and Z ₁₀ is absent; or i) Z ₁ represents G; Z ₂ represents S; Z ₃ represents K; Z ₄ represents S; Z ₅ represents D; Z ₆ represents H; Z ₇ represents S; Z ₈ represents M; Z ₉ missing and Z ₁₀ missing; or j) Z ₁ represents G; Z ₂ represents S; Z ₃ represents K; Z ₄ represents S; Z ₅ represents D; Z ₆ represents H; Z ₇ represents S; Z ₈ missing; Z ₉ missing and Z ₁₀ missing; or k) Z ₁ is missing; Z ₂ represents S; Z ₃ represents K; Z ₄ represents S; Z ₅ represents D; Z ₆ represents H; Z ₇ represents S; Z ₈ represents M; Z ₉ missing and Z ₁₀ missing; or l) Z ₁ is missing; Z ₂ represents S; Z ₃ represents K; Z ₄ represents S; Z ₅ represents D; Z ₆ represents H; Z ₇ represents S; Z ₈ missing; Z ₉ missing and Z ₁₀ missing; or m) Z ₁ is missing; Z ₂ is missing; Z ₃ represents K; Z ₄ represents S; Z ₅ represents D; Z ₆ represents H; Z ₇ represents S; Z ₈ represents M; Z ₉ represents S and Z ₁₀ is absent; or n) Z ₁ is missing; Z ₂ is missing; Z ₃ is missing; Z ₄ represents S; Z ₅ represents D; Z ₆ represents H; Z ₇ represents S; Z ₈ represents M; Z ₉ represents S and Z ₁₀ is absent; or o) Z ₁ is missing; Z ₂ is missing; Z ₃ represents K; Z ₄ represents S; Z ₅ represents D; Z ₆ represents H; Z ₇ represents S; Z ₈ missing; Z ₉ missing and Z ₁₀ missing; or p) Z ₁ is missing; Z ₂ is missing; Z ₃ is missing; Z ₄ represents S; Z ₅ represents D; Z ₆ represents H; Z ₇ represents S; Z ₈ represents M; Z ₉ is missing and Z ₁₀ is missing. 6. L-nucleic acid molecule according to any one of paragraphs. 4, 5, where: a) the first terminal nucleotide fragment contains the nucleotide sequence 5'-CUGAC-3', and the second terminal nucleotide fragment contains the nucleotide sequence 5'-GUCAG-3'; b) the first terminal nucleotide fragment contains the nucleotide sequence 5'-GCUGAC-3', and the second terminal nucleotide fragment contains the nucleotide sequence 5'-GUCAGC-3'; or c) the first terminal nucleotide fragment contains the nucleotide sequence 5'-GCUGAC-3', and the second terminal nucleotide fragment contains the nucleotide sequence 5'-GUGAGC-3'; d) the first terminal nucleotide fragment contains the nucleotide sequence 5'-GUGAC-3', and the second terminal nucleotide fragment contains the nucleotide sequence 5'-GUCAC-3'; e) the first terminal nucleotide fragment contains a nucleotide sequence of 5'-UGAC-3', and the second terminal nucleotide fragment contains a nucleotide sequence of 5'-GUCA-3'; f) the first terminal nucleotide fragment contains the nucleotide sequence 5'-GGAC-3', and the second terminal nucleotide fragment contains the nucleotide sequence 5'-GUCC-3'; g) the first terminal nucleotide fragment contains the nucleotide sequence 5'-GCAC-3', and the second terminal nucleotide fragment contains the nucleotide sequence 5'-GUGC-3'; h) the first terminal nucleotide fragment contains the nucleotide sequence 5'-GGUC-3', and the second terminal nucleotide fragment contains the nucleotide sequence 5'-GACC-3'; i) the first terminal nucleotide fragment contains the nucleotide sequence 5'-UGGC-3', and the second terminal nucleotide fragment contains the nucleotide sequence 5'-GCCA-3'; j) the first terminal nucleotide fragment contains the nucleotide sequence 5'-GAC-3', and the second terminal nucleotide fragment contains the nucleotide sequence 5'-GUC-3'; where preferable the first terminal nucleotide fragment contains the nucleotide sequence 5'-CUGAC-3', and the second terminal nucleotide fragment contains the nucleotide sequence 5'-GUCAG-3', or the first terminal nucleotide fragment contains the nucleotide sequence 5'-GCUGAC-3', and the second terminal nucleotide fragment contains the nucleotide sequence 5'-GUCAGC-3', or the first terminal nucleotide fragment contains the nucleotide sequence 5'-GUGAC-3', and the second terminal nucleotide fragment contains the nucleotide sequence 5'-GUCAC-3'; where is more preferable the first terminal nucleotide fragment contains the nucleotide sequence 5'-CUGAC-3', and the second terminal nucleotide fragment contains the nucleotide sequence 5'-GUCAG-3' or the first terminal nucleotide fragment contains the nucleotide sequence 5'-GUGAC-3', and the second terminal nucleotide fragment contains the nucleotide sequence 5'-GUCAC-3'. 7. L-nucleic acid molecule according to any one of paragraphs. 1-6, wherein the L-nucleic acid molecule contains from 40 to 45 nucleotides, preferably from 42 to 45 nucleotides, and more preferably 42 nucleotides. 8. L-nucleic acid molecule according to any one of paragraphs. 1-7, where the L-nucleic acid molecule consists of ribonucleotides. 9. L-nucleic acid molecule according to any one of paragraphs. 1-8, wherein the L-nucleic acid molecule contains a nucleotide sequence selected from the group of SEQ ID NOs: 14-26 and 39-44, or the L-nucleic acid molecule contains an L-nucleic acid molecule that is at least 75% identical an L-nucleic acid molecule containing a nucleotide sequence selected from the group of SEQ ID NOs: 14-26 and 39-44, or the L-nucleic acid molecule contains an L-nucleic acid molecule that is homologous to an L-nucleic acid molecule containing the nucleotide sequence, selected from the group of SEQ ID NOs: 14-26 and 39-44, where such homology is at least 75%. 10. The L-nucleic acid molecule according to claim 9, where the L-nucleic acid molecule contains a nucleotide sequence selected from the group SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 20 and SEQ ID NO: 41-44, or the L-nucleic acid molecule contains an L-nucleic acid molecule that is at least 75% identical to the L-nucleic acid molecule an acid containing a nucleotide sequence selected from the group SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 20 and SEQ ID NO: 41-44, or the L-nucleic acid molecule contains an L-nucleic acid molecule that is homologous to an L-nucleic acid molecule containing a nucleotide sequence selected from the group SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 20 and SEQ ID NO: 41-44, where the homology is at least 75%. 11. L-nucleic acid molecule according to any one of paragraphs. 1-10, wherein the L-nucleic acid molecule has the ability to bind to CXCL8, wherein preferably CXCL8 is human CXCL8, simian CXCL8, rabbit CXCL8, porcine CXCL8, canine CXCL8, ovine CXCL8 or guinea pig CXCL8. 12. L-nucleic acid molecule according to any one of paragraphs. 1-11, wherein the L-nucleic acid molecule has the ability to specifically bind to CXCL8, preferably human CXCL8, simian CXCL8, rabbit CXCL8, porcine CXCL8, canine CXCL8, sheep CXCL8 or guinea pig CXCL8, and more preferably human CXCL8. 13. L-nucleic acid molecule according to any one of paragraphs. 1-12, wherein the L-nucleic acid molecule does not bind or is not capable of binding to ELR-positive CXC chemokines other than CXCL8, wherein the ELR-positive CXC chemokines other than CXCL8 are preferably selected from the group consisting of CXCL1, CXCL2, CXCL3, CXCL5, CXCL6 and CXCL7. 14. The L-nucleic acid molecule of claim 13, wherein the L-nucleic acid molecule does not bind or is not capable of binding to ELR-positive human CXC chemokines other than human CXCL8, wherein the ELR-positive human CXC chemokines other than human CXCL8 , are preferably selected from the group consisting of human CXCL1, human CXCL2, human CXCL3, human CXCL5, human CXCL6 and human CXCL7. 15. L-nucleic acid molecule according to any one of paragraphs. 1-14, wherein the L-nucleic acid molecule has a binding affinity for human CXCL8, expressed as KD, of 10 nM or less, preferably 1 nM or less, more preferably 100 pM or less, and most preferably 30 pM or less. 16. L-nucleic acid molecule according to any one of paragraphs. 1-15, wherein the L-nucleic acid molecule has a binding affinity for human CXCL8, expressed as IC ₅₀ , of 10 nM or less, preferably 1 nM or less, more preferably 100 pM or less, and most preferably 30 pM or less. 17. L-nucleic acid molecule according to any one of paragraphs. 1-16, wherein the L-nucleic acid molecule has a binding affinity for ELR-positive chemokines CXC other than CXCL8, expressed as KD and being 100 nM or more, preferably 500 nM or more, and more preferably 1000 nM or more. 18. L-nucleic acid molecule according to any one of paragraphs. 1-17, wherein the L-nucleic acid molecule has a binding affinity for ELR-positive chemokines CXC other than CXCL8, expressed as IC ₅₀ and being 100 nM or more, preferably 500 nM or more, and more preferably 1000 nM or more. 19. L-nucleic acid molecule according to any one of paragraphs. 1-18, wherein the L-nucleic acid molecule has a binding affinity for human CXCL8, expressed as KD, of 10 nM or less, preferably 1 nM or less, more preferably 100 pM or less, and most preferably 30 pM or less, wherein the molecule The L-nucleic acid has a binding affinity for ELR-positive chemokines CXC other than CXCL8, expressed as KD, of 100 nM or more, preferably 500 nM or more, and more preferably 1000 nM or more, and/or the L-nucleic acid molecule has a binding affinity for human CXCL8, expressed as IC ₅₀ , of 10 nM or less, preferably 1 nM or less, more preferably 100 pM or less, and most preferably 30 pM or less, wherein the L-nucleic acid molecule has a binding affinity for ELR-positive chemokines CXC other than CXCL8, expressed as IC ₅₀ and being 100 nM or more, preferably 500 nM or more, and more preferably 1000 nM or more. 20. L-nucleic acid molecule according to any one of paragraphs. 15-19, where the binding affinity is determined at room temperature, and preferably at 25°C. 21. L-nucleic acid molecule according to any one of paragraphs. 15-19, where binding affinity is determined at 37°C. 22. L-nucleic acid molecule according to any one of paragraphs. 1-21, wherein the L-nucleic acid molecule is capable of inhibiting the actions of CXCL8 on its receptors CXCR1 and/or CXCR2, preferably human CXCL8, simian CXCL8, rabbit CXCL8, porcine CXCL8, canine CXCL8, sheep CXCL8 or guinea pig CXCL8, and more preferably human CXCL8. 23. The L-nucleic acid molecule of claim 22, wherein the L-nucleic acid molecule is not an antagonist of ELR-positive human CXC chemokines other than human CXCL8 or is not capable of inhibiting activity mediated by ELR-positive CXC chemokines other than CXCL8, wherein the ELR-positive CXC chemokines other than CXCL8 are preferably selected from the group consisting of CXCL1, CXCL2, CXCL3, CXCL5, CXCL6 and CXCL7. 24. The L-nucleic acid molecule of claim 23, wherein the L-nucleic acid molecule is not an antagonist of ELR-positive human CXC chemokines other than human CXCL8 or is not capable of inhibiting activity mediated by ELR-positive CXC chemokines other than CXCL8, wherein the ELR-positive human CXC chemokines other than human CXCL8 are preferably selected from the group consisting of human CXCL1, human CXCL2, human CXCL3, human CXCL5, human CXCL6 and human CXCL7. 25. L-nucleic acid molecule according to any one of paragraphs. 22-24, wherein the antagonistic activity of the L-nucleic acid molecule relative to the activity mediated by human CXCR1 and/or CXCR2, expressed as _IC50 , is 10 nM or less, preferably 1 nM or less, more preferably 100 pM or less, and most preferably 30 pM or less. 26. L-nucleic acid molecule according to any one of paragraphs. 22-25, wherein the antagonistic activity of the L-nucleic acid molecule to that mediated by human CXCL8 is determined at 37°C. 27. L-nucleic acid molecule according to any one of paragraphs. 1-26, where the modifying group is attached to the 5'-terminal nucleotide or 3'-terminal nucleotide of the L-nucleic acid molecule. 28. L-nucleic acid molecule according to any one of paragraphs. 1-27, where the modifying group is attached to the L-nucleic acid molecule via a linker. 29. The L-nucleic acid molecule of claim 28, wherein the linker is a hydrophilic linker, wherein the hydrophilic linker preferably contains one or more ethylene glycol units, and more preferably the hydrophilic linker contains triethylene glycol, hexaethylene glycol or polyethylene glycol. 30. The L-nucleic acid molecule according to claim 28, wherein the linker is a biodegradable linker. 31. L-nucleic acid molecule according to any one of paragraphs. 1-30, where the rate of excretion of an L-nucleic acid molecule containing a modifying group from the body is reduced compared to the rate of excretion of an L-nucleic acid molecule not containing a modifying group. 32. L-nucleic acid molecule according to any one of paragraphs. 1-30, wherein the L-nucleic acid molecule containing the modifying group has a longer retention time in the body compared to the retention time in the body of the L-nucleic acid molecule not containing the modifying group. 33. L-nucleic acid molecule according to any one of paragraphs. 31, 32, where the organism is a human or animal organism, and preferably a human organism. 34. L-nucleic acid molecule according to any one of paragraphs. 1 and 33, wherein the modifying group is selected from the group consisting of a biodegradable modification and a non-biodegradable modification, and wherein the modifying group is selected from the group consisting of polyethylene glycol, straight chain polyethylene glycol, branched polyethylene glycol and hydroxyethyl starch. 35. The L-nucleic acid molecule of claim 34, wherein the modifying group is a polyethylene glycol, preferably a straight chain or branched polyethylene glycol, wherein the molecular weight of the polyethylene glycol is preferably from about 20,000 to about 120,000 Da, more preferably from about 30,000 to about 80,000 Yes, and most preferably approximately 40,000 Yes. 36. The L-nucleic acid molecule of claim 35, wherein the molecular weight of the polyethylene glycol is from about 20,000 to about 120,000 Daltons, preferably from about 30,000 to about 80,000 Daltons, and most preferably about 40,000 Daltons. 37. The L-nucleic acid molecule of claim 34, wherein the modifying group is hydroxyethylated starch, wherein the molecular weight of the hydroxyethylated starch is preferably from about 50 to about 1000 kDa, more preferably from about 100 to about 700 kDa, and most preferably from 200 up to 500 kDa. 38. L-nucleic acid molecule according to any one of paragraphs. 1-30, where the modifying group is intended to immobilize the L-nucleic acid molecule. 39. The L-nucleic acid molecule of claim 38, wherein immobilization comprises covalently binding the L-nucleic acid molecule to a surface, wherein the preferred surface is the surface of a reaction vessel, the surface of a device in the reaction vessel, or the surface of a biosensor. 40. The L-nucleic acid molecule of claim 38, wherein immobilization comprises non-covalently binding the L-nucleic acid molecule to a surface, where the preferred surface is the surface of a reaction vessel, the surface of a device in the reaction vessel, or the surface of a biosensor. 41. L-nucleic acid molecule according to any one of paragraphs. 38-40, wherein the modifying group is selected from the group consisting of carboxyl, hydroxyl, phosphatidyl, sulfonic acid ester, amine, thiol, epoxide, alkyne, steric cycloalkyne, maleimide, azide, hydrazide, wherein the steric cycloalkyne is preferably selected from the group consisting of dibenzocyclooctyl (DBCO), bicyclo[6.1.0]non-4-yne (BCN) and azadibenzocyclooctine (ADIBO). 42. The L-nucleic acid molecule according to claim 41, wherein the modifying group is a steric cycloalkyne, and preferably dibenzocyclooctyl (DBCO). 43. The L-nucleic acid molecule according to claim 40, wherein the modifying group is biotin, bromodeoxyuridine, digoxigenin or oligonucleotide, and preferably the modifying group is biotin. 44. L-nucleic acid molecule according to any one of paragraphs. 1-30, wherein the modifying group allows detection of the L-nucleic acid molecule, and preferably the modifying group is a label. 45. The L-nucleic acid molecule according to claim 44, where the label is selected from the group consisting of biotin, bromodeoxyuridine, digoxigenin, oligonucleotide, fluorescent label, electrochemiluminescent label, radioactive label, enzyme label, UV label, colloidal gold, nanoparticle and label in form of a molecule forming a chelate complex. 46. Use of an L-nucleic acid molecule according to any one of paragraphs. 1-45 in the treatment or prevention of a disease, wherein the disease is associated with a CXCL8-mediated pathogenic mechanism and is selected from the group consisting of an inflammatory disease and cancer, wherein the inflammatory disease is preferably an inflammatory disease of the airways, an inflammatory skin disease, an autoimmune disease, an inflammatory nerve disease , ischemic disease, atherosclerosis, pancreatic islet graft rejection, organ transplant rejection, delayed organ function, spinal cord injury, or cystitis. 47. A pharmaceutical composition containing an L-nucleic acid molecule according to any one of paragraphs. 1-45, in an effective amount for the treatment or prevention of a disease that is associated with a CXCL8-mediated pathogenic mechanism and is selected from the group consisting of an inflammatory disease and cancer, wherein the inflammatory disease is preferably an inflammatory disease of the respiratory tract, an inflammatory disease of the skin, an autoimmune disease, inflammatory nervous disease, ischemic disease, atherosclerosis, pancreatic islet graft rejection, organ transplant rejection, delayed organ function, spinal cord injury or cystitis, wherein the composition optionally contains an additional component, wherein the additional component is selected from the group consisting of a pharmaceutically acceptable excipient, a pharmaceutically acceptable carrier and a pharmaceutically active agent. 48. The pharmaceutical composition according to claim 47, where the pharmaceutical composition contains an L-nucleic acid molecule according to any one of paragraphs. 1-45 and a pharmaceutically acceptable carrier. 49. Use of an L-nucleic acid molecule according to any one of paragraphs. 1-45 for the preparation of a medicament for the treatment or prevention of a disease that is associated with a CXCL8-mediated pathogenic mechanism and is selected from the group consisting of an inflammatory disease and cancer, wherein the inflammatory disease is preferably an inflammatory disease of the respiratory tract, an inflammatory disease of the skin, an autoimmune disease, inflammatory nerve disease, ischemic disease, atherosclerosis, pancreatic islet graft rejection, organ transplant rejection, delayed organ function, spinal cord injury, or cystitis. 50. Use according to claim 49, where the medicinal product is intended for use in medicine or veterinary medicine. 51. Use of an L-nucleic acid molecule according to any one of paragraphs. 1-45 to obtain a diagnostic agent, diagnostic tool or biosensor for detecting CXCL8. 52. Use according to claim 51, wherein the diagnostic agent or diagnostic agent has a surface on the reaction vessel or a surface on a device in the reaction vessel. 53. Application according to claim 51, where the biosensor has a surface. 54. Application according to any one of paragraphs. 51-53, where CXCL8 is human CXCL8. 55. Application according to any one of paragraphs. 51-52, wherein CXCL8 is detected by direct binding, a sandwich binding assay, or a competitive binding assay. 56. Use according to claim 55, where direct binding includes a detection method in which only the L-nucleic acid molecule according to any one of claims is used. 1-45 and which does not use or which does not include the use of a second molecule different from the L-nucleic acid molecule of any one of paragraphs. 1-45, which interacts with CXCL8. 57. Use according to claim 55, where the sandwich assay is a sandwich hybridization assay, in which the L-nucleic acid molecule according to any one of claims. 1-45 is immobilized on the surface, and the L-nucleic acid molecule complex according to any one of paragraphs. 1-45 and CXCL8 are detected using a detection agent that binds to the epitope(s) of CXCL8 that are different from the epitope(s) associated with the L-nucleic acid molecule of any one of claims. 1-45. 58. The use of claim 55, wherein the sandwich assay is a sandwich hybridization assay, wherein an agent that binds to a CXCL8 epitope(s) that is different from the epitope(s) bound with an L-nucleic acid molecule according to any one of paragraphs. 1-45 are immobilized on the surface, and the complex of such agents and CXCL8 is detected using the L-nucleic acid molecule according to any one of paragraphs. 1-45. 59. Use according to claim 55, where in the competitive binding assay the L-nucleic acid molecule according to any one of claims. 1-45 is immobilized on the surface and binding of the L-nucleic acid molecule according to any one of paragraphs. 1-45 with CXCL8 competes with oligonucleotide probes complementary to the L-nucleic acid molecule according to any one of paragraphs. 1-45, or with CXCL8, where the complementarity is preferably base complementarity, and more preferably is based on Watson-Crick base pairing. 60. Use according to claim 55, where in the competitive binding assay the L-nucleic acid molecule according to any one of claims. 1-45 are not immobilized on the surface and binding of the L-nucleic acid molecule according to any one of paragraphs. 1-45 with CXCL8 competes with oligonucleotide probes complementary to the L-nucleic acid molecule according to any one of paragraphs. 1-45, or with CXCL8, where preferably oligonucleotide probes complementary to the L-nucleic acid molecule according to any one of paragraphs. 1-45 or CXCL8 is immobilized on a surface where the complementarity is preferably base complementarity, and more preferably such complementarity is based on Watson-Crick base pairing. 61. Application according to any one of paragraphs. 57-60, where an L-nucleic acid molecule according to any one of paragraphs is used for immobilization on the surface. 38-43. 62. Application according to any one of paragraphs. 52-61, where the surface, and preferably the surface of the biosensor, is selected from the group consisting of gold, silver, titanium, zirconium, vanadium, chromium, manganese, cobalt, tungsten, molybdenum, platinum, aluminum, iron, steel, copper, nickel, silicon, germanium, indium phosphide, gallium arsenide and oxide, nitride or alloys or mixtures thereof, indium tin oxide, glassy carbon, sapphire, silicate glass and borate glass. 63. Application according to any one of paragraphs. 52-62, where the surface, and preferably the surface of the biosensor, is modified with polylysine; aminosilane; epoxysilane; nitrocellulose; carboxydextran; carbon nanomembrane; graphene oxide; carbon surface such as black carbon, carbon fiber, carbon plate, carbon cloth, activated carbon, glassy carbon, carbon, activated carbon, graphite powder, graphite fiber, carbon nanotube; fullerene; carboxyl group; an azide group; thiol group; hydroxy group; epoxide; maleimide; alkyne; steric alkyne or any combination thereof. 64. Use according to claim 63, where for immobilization of an L-nucleic acid molecule according to any one of claims. 38-43, the surface is functionalized with a primary amine, thiol group, azide, alkyne, alkene, tetrazine, tetrazole or steric cycloalkyne, where the steric cycloalkyne is preferably selected from the group consisting of dibenzocyclooctyl (DBCO), bicyclo[6.1.0]non-4-yne (BCN) or azadibenzocyclooctine (ADIBO). 65. Use according to claim 64, where for immobilization of an L-nucleic acid molecule according to any one of claims. 41, 42 perform an azide–alkyne cycloaddition reaction promoted by a steric alkyne. 66. Application according to any one of paragraphs. 51-65, where the L-nucleic acid molecule according to any one of paragraphs. 43-45 are used for detection. 67. Application according to any one of paragraphs. 51-66, where the detection system of the biosensor is based on optical sensing, mass change, refractive index, charge, surface tension and interatomic force based microscopy. 68. Use according to claim 67, wherein the biosensor detection system based on optical sensing is fluorescence, absorption or luminescence. 69. The use of claim 67, wherein the biosensor detection system based on mass change is a quartz crystal microbalance system or microelectromechanical systems. 70. The use of claim 67, wherein the refractive index based biosensor detection system is surface plasmon resonance, circular resonator or ellipsometry. 71. The use of claim 67, wherein the charge-based detection system of the biosensor is electrochemical resistance, voltammetry, amperometry, potentiometry, conductance, or a field-effect transistor. 72. Use according to claim 67, wherein the biosensor detection system based on surface tension is a cantilever biosensor. 73. Application according to any one of paragraphs. 51-54, wherein CXCL8 is detected using a homogeneity assay. 74. Use of an L-nucleic acid molecule according to any one of paragraphs. 1-45 for detecting CXCL8, and preferably human CXCL8. 75. Use according to claim 74, wherein CXCL8 is detected using a sandwich binding assay, a homogeneity assay, or a competitive binding assay. 76. The use of claim 75, wherein the sandwich binding assay, homogeneity assay, or competitive binding assay is suitable for direct or indirect detection of CXCL8. 77. Application according to any one of paragraphs. 73-76, where the L-nucleic acid molecule is covalently immobilized on the surface of a reaction vessel with a diagnostic agent, on the surface of a device in a reaction vessel with a diagnostic agent, or on the surface of a biosensor. 78. Application according to claim 77, where an L-nucleic acid molecule according to any one of claims is used for immobilization. 38-43. 79. Application according to any one of paragraphs. 77, 78, where the surface, and preferably the surface of the biosensor, is selected from the group consisting of gold, silver, titanium, zirconium, vanadium, chromium, manganese, cobalt, tungsten, molybdenum, platinum, aluminum, iron, steel, copper, nickel, silicon , germanium, indium phosphide, gallium arsenide and oxide, nitride or alloys or mixtures thereof, indium tin oxide, glassy carbon, sapphire, silicate glass and borate glass. 80. Application according to any one of paragraphs. 77-79, where the surface, and preferably the surface of the biosensor, is modified with polylysine; aminosilane; epoxysilane; nitrocellulose; carboxydextran; carbon nanomembrane; graphene oxide; carbon surface such as black carbon, carbon fiber, carbon plate, carbon cloth, activated carbon, glassy carbon, carbon, activated carbon, graphite powder, graphite fiber, carbon nanotube; fullerene; carboxyl group; an azide group; thiol group; hydroxy group; epoxide; maleimide; alkyne; steric alkyne or any combination thereof. 81. Use according to claim 80, where for immobilization of an L-nucleic acid molecule according to any one of claims. 38-43 the surface is functionalized with an activated primary amine, a thiol group, an azide or a steric cycloalkyne, where the steric cycloalkyne is preferably selected from the group consisting of dibenzocyclooctyl (DBCO), bicyclo[6.1.0]non-4-yne (BCN) or azadibenzocyclooctyl (ADIBO) . 82. Use according to claim 81, where for immobilization of an L-nucleic acid molecule according to any one of claims. 41, 42 perform an azide–alkyne cycloaddition reaction promoted by a steric alkyne. 83. Application according to any one of paragraphs. 77-82, where the detection system of the biosensor is based on optical sensing, mass change, refractive index, charge, surface tension and interatomic force based microscopy. 84. Use according to claim 83, wherein the biosensor detection system based on optical sensing is fluorescence, absorption or luminescence. 85. The use of claim 80, wherein the biosensor detection system based on mass change is a quartz crystal microbalance system or a microelectromechanical system. 86. The use of claim 83, wherein the refractive index based biosensor detection system is surface plasmon resonance, circular resonator or ellipsometry. 87. The use of claim 83, wherein the charge-based detection system of the biosensor is electrochemical resistance, voltammetry, amperometry, potentiometry, conductance, or field-effect transistor. 88. Use according to claim 83, wherein the biosensor detection system based on surface tension is a cantilever biosensor. 89. Application according to any one of paragraphs. 74-76, wherein the L-nucleic acid molecule contains a modifying group that allows the L-nucleic acid molecule to be detected, and preferably the modifying group is a label. 90. Use according to claim 89, where the label is selected from the group consisting of biotin, bromodeoxyuridine, digoxigenin, oligonucleotide, fluorescent label, electrochemiluminescent label, radioactive label, enzyme label, UV label, colloidal gold, nanoparticle and label in the form of a molecule forming chelate complex. 91. A kit for detecting CXCL8, where the kit contains an L-nucleic acid molecule according to any one of paragraphs. 1-45 and at least an instruction leaflet or reaction vessel. 92. A method for detecting CXCL8 using L-nucleic acid according to any one of claims. 1-45 in a sample, wherein said method includes the steps of: a) obtaining a sample with an unknown concentration of CXCL8; b) contacting the sample or diluting it with a diagnostic agent, diagnostic tool or biosensor according to any one of paragraphs. 51-73; c) measuring the signal using a diagnostic agent, diagnostic tool or biosensor according to any one of claims. 51-73; d) comparing, optionally, the signal with a reference; e) wherein, optionally, the concentration of CXCL8 in a sample with an unknown concentration of CXCL8 is determined by comparison with signals obtained from at least one sample with a known concentration of CXCL8, and preferably at least one sample with a known concentration of CXCL8 is subjected to steps b)-c) . 93. A complex containing an L-nucleic acid molecule according to any one of paragraphs. 1-45 and CXCL8 to immobilize CXCL8 on a surface, where the complex is preferably a crystalline complex. 94. A method of screening a compound for the ability to inhibit the actions of CXCL8 on its receptors CXCR1 and/or CXCR2, where the method includes the following steps: - obtaining a candidate connection, - obtaining an L-nucleic acid molecule according to any one of claims. 1-45; - creation of a test system that will transmit a signal in the presence of a compound capable of inhibiting the actions of CXCL8 on its receptors CXCR1 and/or CXCR2, and - determining whether the candidate compound is a compound capable of inhibiting the actions of CXCL8 on its receptors CXCR1 and/or CXCR2. 95. A method for detecting L-nucleic acid according to any one of claims. 1-45 in a sample, wherein said method includes the steps of: a) obtaining a capture probe, wherein the capture probe is at least partially complementary to the first part of the L-nucleic acid molecule according to any one of claims. 1-45, and a detection probe, wherein the detection probe is at least partially complementary to the second part of the L-nucleic acid molecule according to any one of paragraphs. 1-45; or, alternatively, wherein the capture probe is at least partially complementary to the second portion of the L-nucleic acid molecule according to any one of claims. 1-45, and the detection probe is at least partially complementary to the first part of the L-nucleic acid molecule according to any one of paragraphs. 1-45; b) adding a capture probe and a detection probe, alone or in combination, to the sample containing the L-nucleic acid molecule according to any one of claims. 1-45, or presumably containing an L-nucleic acid molecule according to any one of paragraphs. 1-45; c) allowing the capture probe and the detection probe to interact, either simultaneously or sequentially in any order, with the L-nucleic acid molecule according to any one of claims. 1-45, or parts thereof; d) optionally, determining whether the capture probe hybridizes with the L-nucleic acid molecule of any one of claims. 1-45, as described in step a); And e) detecting the complex obtained in step c) and consisting of an L-nucleic acid molecule according to any one of paragraphs. 1-45, capture probe and detection probe. 96. The method of claim 95, wherein the detection probe comprises a detection agent and/or wherein the capture probe is immobilized on a support, preferably a solid support. 97. The method of claim 95 or 96, wherein any detection probe that is not part of the complex obtained in step c) is removed from the reaction mixture so that only the detection probe that is part of the complex is detected in step e). 98. The method according to any one of paragraphs. 95-97, where step e) includes the step of comparing the signal generated by the detection means if the capture probe and the detection probe hybridize in the presence of an L-nucleic acid molecule according to any one of claims. 1-45 or parts thereof and in the absence of said L-nucleic acid molecule according to any one of paragraphs. 1-45.

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