RU2798978C2 - Radiopharmaceuticals targeting grpr and their use - Google Patents

Abstract

FIELD: radiopharmaceuticals.

SUBSTANCE: invention relates to gastrin-releasing peptide (GRPR) receptor-targeted pharmaceutical composition containing a radioactively labeled GRPR antagonist and a surfactant containing polyethylene glycol-15-hydroxystearate. The antagonist has the following formula: MС-S-P, where M is a radioactive metal and C is a chelator that binds M; S is an optional spacer covalently linked between the C and N-terminus of P; P is a peptide GRP receptor antagonist with the general formula: Xaa1-Xaa2-Xaa3-Xaa4-Xaa5-Xaa6-Xaa7-Z, where Xaa1 is the D-isomer of Phe; Xaa2 is Gln; Xaa3 is Trp; Xaa4 is Ala; Xaa5 is Val; Xaa6 is Gly; Xaa7 is His; Z is -NH-alkyl or Z is

, where X is NH, and R1 and R2 are the same or different and are selected from proton or optionally substituted alkyl. The invention also relates to the use of the said pharmaceutical composition, a method for treating or preventing cancer, and a method for imaging a tumor.

EFFECT: invention makes it possible to obtain a stable pharmaceutical composition by preventing peptide adhesion.

13 cl, 8 dwg, 3 tbl, 2 ex

Description

The field of technology to which the invention belongs

The present invention relates to radiopharmaceuticals directed to the gastrin-releasing peptide receptor (GRPR) and their use. In particular, the present invention relates to a pharmaceutical composition containing a radioactively labeled GRPR antagonist and a surfactant. The present invention also relates to a radiolabeled GRPR antagonist for use in the treatment or prevention of cancer.

Prior Art

The gastrin-releasing peptide receptor (GRPR), also known as the bombesin subtype 2 receptor, is a G-protein coupled receptor expressed in various organs, including the gastrointestinal tract and pancreas (Guo M, et al. Curr Opin Endocrinol Diabetes Obes 2015; 22:3-8.2; Gonzalez N, et al. Curr Opin Enocrinol Diabetes Obes 2008; 15:58-64). Upon binding of the appropriate ligand, GRPR is activated, inducing multiple physiological processes such as the regulation of exocrine and endocrine secretion (Guo M, et al. Curr Opin Endocrinol Diabetes Obes. 2015; 22:3–8.2; Gonzalez N, et al. Curr Opin Enocrinol Diabetes Obes 2008; 15:58–64). In recent decades, GRPR expression has been reported in various types of cancer, including prostate cancer and breast cancer (Gugger M and Reubi JC. Gastrin-releasing peptide receptors in non-neoplastic and neoplastic human breast. Am J Pathol. 1999; 155:2067– 2076; Markwalder R and Reubi JC. Cancer Res. 1999; 59:1152–1159). Thus, GRPR has become an interesting target for receptor-mediated tumor imaging and treatment, such as peptide receptor scintigraphy and peptide receptor radionuclide therapy (Gonzalez N, et al. Curr Opin Enocrinol Diabetes Obes. 2008; 15:58–64). Following the successful application of radiolabeled peptide analogs of somatostatin in neuroendocrine tumors for nuclear imaging and therapy (Brabander T, et al. Front Horm Res. 2015; 44:73–87; Kwekkeboom DJ and Krenning EP. Hematol Oncol Clin North Am. 2016; 30:179–191), a variety of radiolabeled GRPR radioligands have been synthesized and studied in preclinical as well as clinical studies, mainly in patients with prostate cancer. Examples of such peptide analogs include AMBA, the Demobesin series, and MP2653 (Yu Z, et al. Curr Pharm Des. 2013; 19:3329-3341; Lantry LE, et al. J Nucl Med. 2006; 47:1144-1152.; Schroeder RP et al Eur J Nucl Med Mol Imaging 2010 37:1386–1396 Nock B et al Eur J Nucl Med Mol Imaging 2003 30:247–258 Mather SJ et al Mol Imaging Biol 2014; 16:888–895). Recent studies have shown superiority of GRPR antagonists over GRPR agonists (Mansi R, et al. Eur J Nucl Med Mol Imaging. 2011; 38:97–107; Cescato R, et al. J Nucl Med. 2008; 49:318–326). Compared to receptor agonists, antagonists often show higher binding and favorable pharmacokinetics (Ginj M, et al. Proc Natl Acad Sci USA. 2006; 103:16436-16441). In addition, clinical studies with radiolabeled GRPR agonists have shown undesirable side effects in patients caused by GRPR activation after peptide binding to the receptor ((Bodei L, et al. [abstract]. Eur J Nucl Med Mol Imaging. 2007; 34:S221 ).

It has recently been discovered that certain GRPR antagonists, such as NeoBOMB1, can be labeled with various radionuclides and can potentially be used for imaging and treatment of GRPR-expressing cancers, such as, but not limited to, prostate cancer and breast cancer. However, only biodistribution studies have been reported to date, and no effective treatment protocols or pharmaceutical formulations have been developed.

Thus, in this context, it is necessary to provide a pharmaceutical composition comprising a GRPR antagonist that can be used in patients. In addition, it is also necessary to provide an effective protocol for the treatment of cancer patients using a GRPR antagonist.

Statement of the Invention

In a first aspect, the present invention relates to a pharmaceutical composition comprising:

a radioactively labeled GRPR antagonist having the following formula:

MS-S-P

Where:

M is a radioactive metal and C is a chelator that binds M;

S is an optional spacer covalently linked between the C and N-terminus of P;

P is a peptide GRP receptor antagonist with the general formula:

Xaa1-Xaa2-Xaa3-Xaa4-Xaa5-Xaa6-Xaa7-Z;

Xaa1 is absent or selected from the group consisting of amino acid residues Asn, Thr, Phe, 3-(2-thienyl)alanine (Thi), 4-chlorophenylalanine (Cpa), α-naphthylalanine (α-Nal), β-naphthylalanine (β -Nal), 1,2,3,4-tetrahydronorharman-3-carboxylic acid (Tpi), Tyr, 3-iodo-tyrosine (o-I-Tyr), Trp and pentafluorophenylalanine (5-F-Phe) (all in the form of L- or D-isomers);

Xaa2 is Gln, Asn or His;

Xaa3 is Trp or 1,2,3,4-tetrahydronorharman-3-carboxylic acid (Tpi);

Xaa4 is Ala, Ser or Val;

Xaa5 is Val, Ser or Thr;

Xaa6 is Gly, sarcosine (Sar), D-Ala, or β-Ala;

Xaa7 is His or (3-methyl)histidine (3-Me)His;

Z is selected from -NHOH, -NHNH ₂ , -NH-alkyl, -N(alkyl) ₂ , and -O-alkyl,

or Z is

where X is NH (amide) or O (ester) and R1 and R2 are the same or different and are selected from proton, optionally substituted alkyl, optionally substituted alkyl ether, aryl, aryl ether or alkyl-, halo-, hydroxyl - or a hydroxyalkyl-substituted aryl or heteroaryl group; And

- a surfactant containing a compound having (i) a polyethylene glycol chain and (ii) a fatty acid ester.

In a second aspect, the present invention provides a composition comprising a radiolabeled GRPR antagonist for use in the treatment or prevention of cancer in a subject, wherein:

- a radioactively labeled GRPR antagonist has the following formula:

MS-S-P

Where:

M is a radioactive metal and C is a chelator that binds M;

S is an optional spacer covalently linked between the C and N-terminus of P;

P is a peptide GRP receptor antagonist with the general formula:

Xaa1-Xaa2-Xaa3-Xaa4-Xaa5-Xaa6-Xaa7-Z;

Xaa1 is absent or selected from the group consisting of amino acid residues Asn, Thr, Phe, 3-(2-thienyl)alanine (Thi), 4-chlorophenylalanine (Cpa), α-naphthylalanine (α-Nal), β-naphthylalanine (β -Nal), 1,2,3,4-tetrahydronorharman-3-carboxylic acid (Tpi), Tyr, 3-iodo-tyrosine (o-I-Tyr), Trp and pentafluorophenylalanine (5-F-Phe) (all in the form of L- or D-isomers);

Xaa2 is Gln, Asn or His;

Xaa3 is Trp or 1,2,3,4-tetrahydronorharman-3-carboxylic acid (Tpi);

Xaa4 is Ala, Ser or Val;

Xaa5 is Val, Ser or Thr;

Xaa6 is Gly, sarcosine (Sar), D-Ala, or β-Ala;

Xaa7 is His or (3-methyl)histidine (3-Me)His;

Z is selected from -NHOH, -NHNH ₂ , -NH-alkyl, -N(alkyl) ₂ , and -O-alkyl,

or Z is

,

,

where X is NH (amide) or O (ester) and R1 and R2 are the same or different and are selected from proton, optionally substituted alkyl, optionally substituted alkyl ether, aryl, aryl ether or alkyl-, halo-, hydroxyl - or a hydroxyalkyl-substituted aryl or heteroaryl group; And

the radiolabeled GRPR antagonist is administered to said subject in a therapeutically effective amount of 2,000 to 10,000 MBq.

Brief description of drawings/figures

Figure 1A. Figure 1A shows SPECT/CT images 4 hours and 24 hours after the 1st injection, and 4 hours after the 2nd and 3rd injections. The arrows indicate the tumor. Animals were injected with 30 MBq/300 pmol (group 1), 40 MBq/400 pmol (group 2) or 60 MBq/600 pmol ¹⁷⁷ LU-NeoBOMB1.

Figure 1B. Figure 1B shows quantitative tumor uptake (n=2 per group) after the injections described in Figure 1A.

Figure 2A, 2B. Figure 2A shows extrapolated tumor sizes of untreated animals and animals treated with 3x30 MBq/300 pmol (Group 1), 3x40 MBq/400 pmol (Group 2) and 3x60 MBq/600 pmol ¹⁷⁷ LU -NeoBOMB1 (group 3). Figure 2B shows the survival of untreated animals and animals treated with 3×30 MBq/300 pmol (group 1), 3×40 MBq/400 pmol (group 2) and 3×60 MBq/600 pmol ¹⁷⁷ LU-NeoBOMB1 (group 3).

Figure 3A, 3B. Figure 3A shows the body weight of the animals before treatment, and up to 12 weeks after treatment. Figure 3B shows the body weight of the animals before treatment, and up to 24 weeks after treatment.

Figure 4. Figure 4 shows pancreatic tissue samples after staining with hematoxylin and eosin, from untreated and treated animals (3x30 MBq/300 pmol (Group 1), 3x40 MBq/400 pmol (Group 2 ) and 3×60 MBq/600 pmol ¹⁷⁷ LU-NeoBOMB1 (group 3)).

Figure 5. Figure 5 shows kidney tissue samples stained with hematoxylin and eosin from untreated and treated animals (3x30 MBq/300 pmol (Group 1), 3x40 MBq/400 pmol (Group 2) and 3×60 MBq/600 pmol 177LU-NeoBOMB1 (group 3)). The circled area indicates lesions with lymphocytic infiltration (ID: D, 814, 861, 868 and 862) or with atrophy and fibrosis (ID: 864).

Detailed description of the invention

Definitions

The term "treatment" includes the improvement or cessation of a disease, disorder, or symptom thereof.

The expression "prevention" includes preventing the onset of a disease, disorder, or symptom thereof.

In accordance with the international system of units, "MBq" is the designation for the unit of radioactivity "Megabecquerel".

Used in this application, the abbreviation "PET" means positron emission tomography.

As used in this application, the abbreviation "SPECT" means single photon emission computed tomography.

As used herein, the terms "effective amount" or "therapeutically effective amount" of a compound refer to an amount of a compound that will elicit a biological or medical response in a subject, such as ameliorating symptoms, alleviating conditions, slowing or delaying the progression of a disease, or preventing disease.

As used herein, the terms "substituted" or "optionally substituted" refers to a group that is optionally substituted with one or more substituents selected from: halo, -OR', -NR'R", -SR', -SiR'R"R '”, -OC(O)R', -C(O)R', -CO ₂ R', -C(O)NR'R”, -OC(O)NR'R”, -NR”C( O)R', -NR'-C(O)NR”R'”, -NR”C(O)OR', -NR-C(NR'R”R'”)=NR””, -NR- C(NR'R”)=NR'” -S(O)R', -S(O) ₂ R', -S(O) ₂ NR'R”, -NRSO ₂ R', -CN, -NO ₂ , -R', -N ₃ , -CH(Ph) ₂ , fluoro(C ₁ -C ₄ )alkokso, and fluoro(C ₁ -C ₄ )alkyl in an amount from zero to the total number of open valences on the aromatic ring system ; and where R', R", R'" and R"" can be independently selected from hydrogen, alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl. When a compound of the invention includes more than one R group, for example, each of the R groups is independently selected where they are R', R”, R'” and R”” groups when more than one of these groups is present.

Used in this application, the term "alkyl", as such or as part of another substituent, refers to a linear or branched alkyl functional group having from 1 to 12 carbon atoms. Suitable alkyl groups include methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl and t-butyl, pentyl and its isomers (e.g., n-pentyl, iso-pentyl), and hexyl and its isomers (eg n-hexyl, iso-hexyl).

As used herein, the term "heteroaryl" refers to a polyunsaturated aromatic ring system having a single ring or multiple aromatic rings fused together or covalently bonded, containing from 5 to 10 atoms, where at least one ring is aromatic and at least one the ring atom is a heteroatom selected from N, O, and S. The nitrogen and sulfur heteroatoms may optionally be oxidized, and the nitrogen heteroatoms may optionally be quaternized. Such rings may be fused to an aryl, cycloalkyl or heterocyclyl ring. Non-limiting examples of such heteroaryl include furanyl, thiophenyl, pyrrolyl, pyrazolyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, triazolyl, oxadiazolyl, thiadiazolyl, tetrazolyl, oxatriazolyl, thiatriazolyl, pyridinyl, pyrimidyl, pyrazinyl, pyridazinyl, oxazinyl , dioxynyl, thiazinyl, triazinyl , indolyl, isoindolyl, benzofuranyl, isobenzofuranyl, benzothiophenyl, isobenzothiophenyl, indazolyl, benzimidazolyl, benzoxazolyl, purinyl, benzothiadiazolyl, quinolinyl, isoquinolinyl, cinnolinyl, quinazolinyl, and quinoxalinyl.

As used herein, the terms "aryl" refers to a polyunsaturated aromatic hydrocarbyl group having a single ring or multiple aromatic rings fused together containing 6 to 10 ring atoms, where at least one ring is aromatic. The aromatic ring may optionally include one to two additional rings (cycloalkyl, heterocyclyl or heteroaryl as defined herein) fused to it. Suitable aryl groups include phenyl, naphthyl, and a phenyl ring fused to a heterocyclyl such as benzopyranyl, benzodioxolyl, benzodioxyanyl, and the like.

Used in this application, the term "halogen" refers to fluoro-(-F), chloro-(-Cl), bromo-(-Br), or iodo-(-I) group.

As used herein, the term "optionally substituted aliphatic chain" refers to an optionally substituted aliphatic chain having 4 to 36 carbon atoms, preferably 12 to 24 carbon atoms.

Radioactively labeled GRPR antagonist

As used in this application document, the GRPR antagonist has the following formula:

MS-S-P

Where:

M is a radioactive metal and C is a chelator that binds M;

S is an optional spacer covalently linked between the C and N-terminus of P;

P is a peptide GRP receptor antagonist with the general formula:

Xaa1-Xaa2-Xaa3-Xaa4-Xaa5-Xaa6-Xaa7-Z;

Xaa1 is absent or selected from the group consisting of amino acid residues Asn, Thr, Phe, 3-(2-thienyl)alanine (Thi), 4-chlorophenylalanine (Cpa), α-naphthylalanine (α-Nal), β-naphthylalanine (β -Nal), 1,2,3,4-tetrahydronorharman-3-carboxylic acid (Tpi), Tyr, 3-iodo-tyrosine (o-I-Tyr), Trp and pentafluorophenylalanine (5-F-Phe) (all in the form of L- or D-isomers);

Xaa2 is Gln, Asn or His;

Xaa3 is Trp or 1,2,3,4-tetrahydronorharman-3-carboxylic acid (Tpi);

Xaa4 is Ala, Ser or Val;

Xaa5 is Val, Ser or Thr;

Xaa6 is Gly, sarcosine (Sar), D-Ala, or β-Ala;

Xaa7 is His or (3-methyl)histidine (3-Me)His;

Z is selected from -NHOH, -NHNH ₂ , -NH-alkyl, -N(alkyl) ₂ , and -O-alkyl;

or Z is

where X is NH (amide) or O (ester) and R1 and R2 are the same or different and are selected from proton, optionally substituted alkyl, optionally substituted alkyl ether, aryl, aryl ether, or

alkyl-, halogen-, hydroxyl- or hydroxyalkyl-substituted aryl or heteroaryl group.

According to one embodiment, Z is selected from one of the following formulas, where X is NH or O:

According to one embodiment, chelator C is selected from the group consisting of:

In particular embodiments, C is selected from the group consisting of:

According to one embodiment, S is selected from the group consisting of:

a) aryl-containing residues of the formulas:

,

,

where PABA is P-aminobenzoic acid, PABZA is p-aminobenzylamine, PDA is phenylenediamine, and PAMBZA is (aminomethyl)benzylamine;

b) dicarboxylic acids, ω-aminocarboxylic acids, ω-diaminocarboxylic acids or diamines of the formula:

,

,

where DIG is diglycolic acid and IDA is iminodiacetic acid;

c) PEG based spacers of various chain lengths, in particular PEG spacers selected from:

d) α- and β-amino acids, single or in homologous chains of different lengths, or heterologous chains of different lengths, in particular:

GRP (1-18), GRP (14-18), GRP (13-18), BBN(1-5) or [Tyr4] BB (1-5); or

e) combinations a, b, c and d.

In one embodiment, the GRPR antagonist is selected from the group consisting of compounds of the following formulae:

where MC and P are as defined above.

In one embodiment, P is DPhe-Gln-Trp-Ala-Val-Gly-His-NH-CH(CH ₂ -CH(CH ₃ ) ₂ ) ₂ .

In one embodiment, the radiolabeled GRPR antagonist is NeoBOMB1 radiolabeled of formula (I):

(I)

(I)

(M-DOTA-(p-aminobenzylamine diglycolic acid)- [D-Phe ⁶ , His-NH-CH[(CH ₂ -CH(CH ₃ ) ₂ ] ₂ ¹² , des-Leu ¹³ , des-Met ¹⁴ ] BBN(6-14));

where M is a radioactive metal, preferably M is selected from ¹⁷⁷ Lu, ⁶⁸ Ga and ¹¹¹ In.

In one embodiment, the radiolabeled GRPR antagonist is NeoBOMB2 radiolabeled of formula (II):

(II)

(II)

(MN ₄ (p-aminobenzylamine diglycolic acid)-[D-Phe ⁶ , His-NH-CH[(CH ₂ -CH(CH ₃ ) ₂ ] ₂ ¹² , des-Leu ¹³ , des-Met ¹⁴ ]BBN( 6-14));

where M is a radioactive metal.

In one embodiment, M is a radioactive metal which may be selected from ¹¹¹ In, ^133m In, ^99m Tc, ^94m Tc, ⁶⁷ Ga, ⁶⁶ Ga, ⁶⁸ Ga, ⁵² Fe, ¹⁶⁹ Er, ⁷² As, ⁹⁷ Ru, ²⁰³ Pb , ²¹² Pb, ⁶² Cu, ⁶⁴ Cu, ⁶⁷ Cu, ¹⁸⁶ Re, ¹⁸⁸ Re, ⁸⁶ Y, ⁹⁰ Y, ⁵¹ Cr, ^52m Mn, ¹⁵⁷ Gd, ¹⁷⁷ Lu, ¹⁶¹ Tb, ⁶⁹ Yb, ¹⁷⁵ Yb, ¹⁰⁵ Rh, ¹⁶⁶ Dy, ¹⁶⁶ Ho, ¹⁵³ Sm, ¹⁴⁹ Pm, ¹⁵¹ Pm, ¹⁷² Tm, ¹²¹ Sn, ^117m Sn, ²¹³ Bi, ²¹² Bi, ¹⁴² Pr, ¹⁴³ Pr, ¹⁹⁸ Au, ¹⁹⁹ Au, ⁸⁹ Zr, ²²⁵ Ac, and ⁴⁷ Sc. Preferably M is selected from ¹⁷⁷ Lu, ⁶⁸ Ga and ¹¹¹ In.

In one embodiment, M is ¹⁷⁷ Lu. In this case, a radioactively labeled GRPR antagonist can be used for radionuclide therapy. In another embodiment, M is ⁶⁸ Ga. In this case, a radioactively labeled GRPR antagonist can be used for PET. According to another embodiment, M is ¹¹¹ In. In this case, a radioactively labeled GRPR antagonist can be used for SPECT.

pharmaceutical composition

The GRPR antagonist tends to adhere to glass and plastic surfaces due to non-specific binding (NSB), which is a problem in pharmaceutical formulation. Several surfactants have been tested to ensure a stable formulation. The inventors surprisingly found that among all the surfactants tested, surfactants containing a compound having (i) a polyethylene glycol chain and (ii) a fatty acid ester gave the best results.

In a first aspect, the present invention relates to a pharmaceutical composition comprising a radiolabeled GRPR antagonist described herein and a surfactant containing a compound having (i) a polyethylene glycol chain and (ii) a fatty acid ester. In one embodiment, the surfactant also contains free ethylene glycol.

In one embodiment, the surfactant comprises a compound of formula (III)

(III),

(III)

where n is from 3 to 1000, preferably from 5 to 500, and more preferably from 10 to 50, and

R is a fatty acid chain, preferably an optionally substituted aliphatic chain.

In one embodiment, the surfactant comprises polyethylene glycol 15-hydroxystearate and free ethylene glycol.

The radioactively labeled GRPR antagonist may be present at a concentration providing a volumetric radioactivity of at least 100 MBq/ml, preferably at least 250 MBq/ml. The radioactively labeled GRPR antagonist may be present at a concentration providing a volumetric radioactivity of 100 MBq/ml to 1000 MBq/ml, preferably 250 MBq/ml to 500 MBq/ml.

The surfactant may be present at a concentration of at least 5 μg/ml, preferably at least 25 μg/ml, and more preferably at least 50 μg/ml. The surfactant may be present at a concentration of 5 μg/ml to 5000 μg/ml, preferably 25 μg/ml to 2000 μg/ml, and more preferably 50 μg/ml to 1000 μg/ml.

In one embodiment, the composition contains at least one other pharmaceutically acceptable excipient. The pharmaceutically acceptable excipient may be any of those commonly used, and is limited only by physicochemical considerations such as solubility and lack of reactivity with the active compound(s).

In particular, one or more adjuvants may be selected from stabilizers against radiolytic degradation, buffers, sequestration agents, and mixtures thereof.

As used herein, "anti-radiolytic degradation stabilizer" refers to a stabilizing agent that protects organic molecules against radiolytic degradation, for example, when gamma rays emitted from a radionuclide cleave the bond between the atoms of the organic molecules and radicals are formed, these radicals are then captured by the stabilizer, which avoids any other chemical reactions of these radicals that could lead to unwanted, potentially ineffective, or even toxic molecules. Hence, these stabilizers are also referred to as "free radical scavengers" or "radical scavengers" for short. Other alternative designations for these stabilizers are the terms "radiation stability enhancers", "radiolytic stabilizers", or simply "quenchers".

As used herein, the term "sequestering agent" refers to a chelating agent suitable for complexing the free radionuclide metal ions in the formulation (which do not complex with the radiolabeled peptide).

Buffers include acetate buffer, citrate buffer and phosphate buffer.

According to one embodiment, the pharmaceutical composition is an aqueous solution, such as in an injectable composition. According to a specific embodiment, the pharmaceutical composition is an infusion solution.

Requirements for effective pharmaceutical carriers for injectable compositions are well known to those skilled in the art (see, for example, "Pharmaceutics and Pharmacy Practice", J.B. Lippincott Company, Philadelphia, PA, Banker and Chalmers, eds., pages 238-250 (1982) , and "SHP Handbook on Injectable Drugs", Trissel, 15th ed., pages 622-630 (2009)).

The invention also relates to a method for preparing a pharmaceutical composition containing a combination of a radioactively labeled GRPR antagonist and a surfactant.

The invention also relates to a pharmaceutical composition as described above for use in the treatment or prevention of cancer.

Used in this application, the term "cancer" refers to cells with the ability to autonomous growth, that is, an abnormal status or condition characterized by rapidly proliferating cell growth. Hyperproliferative and neoplastic disease states can be classified as pathological, i.e. characterized by or constituting a disease state, or can be classified as non-pathological, that is, as deviating from the normal state, but not associated with a pathological state. The term is intended to include all types of cancerous growth or oncogenic processes, metastatic tissues or malignantly transformed cells, tissues or organs, regardless of the histopathological type or stage of invasiveness.

In specific embodiments, the cancer is selected from prostate cancer, breast cancer, small cell lung cancer, colon cancer, gastrointestinal stromal tumors, gastrinomas, renal cell carcinomas, gastroenteropancreatic neuroendocrine tumors, esophageal squamous cell carcinomas, neuroblastomas, head and neck squamous cell carcinomas, as well as ovarian, endometrial, and pancreatic tumors displaying neoplasia-associated vasculature, i. GRPR. In one embodiment, the cancer is prostate cancer or breast cancer.

The invention also relates to a pharmaceutical composition as described above for use in in vivo imaging, in particular for detecting GRPR positive tumors in a subject in need thereof, preferably by PET and SPECT imaging.

The invention also relates to a method for treating or preventing cancer in a subject in need thereof, comprising administering to said subject a therapeutically effective amount of a pharmaceutical composition as described above.

The invention also relates to an in vivo imaging method comprising administering to a subject an effective amount of a pharmaceutical composition as described above and detecting a signal derived from the decay of a radioisotope present in said compound.

Radiolabeled GRPR Antagonist for Use in Cancer Treatment

In a second aspect, the invention also provides a composition comprising a radiolabeled GRPR antagonist for use in the treatment or prevention of cancer in a subject in need thereof, wherein the radiolabeled GRPR antagonist is administered to said subject in a therapeutically effective amount of 2,000 to 10,000 MBq. .

In specific embodiments, a therapeutically effective amount of the composition is administered to said subject 2 to 8 times per course of treatment. For example, a patient may be treated with a radioactively labeled GRPR antagonist, such as ¹⁷⁷ Lu-NeoBOMB1, intravenously in 2 to 8 cycles of 2000 to 10,000 mBq each cycle.

In some aspects, the subject is a mammal, such as, but not limited to, a rodent, canine, feline, or primate. In some aspects, the subject is a human.

The inventors have found that ¹⁷⁷ Lu-NeoBOMB1 is effective as shown in animal models of cancer. Compared to untreated animals, the treatment groups had a significantly longer delay in tumor growth and a significantly longer mean survival time. In the non-limiting examples described herein, animals received 3x30 MBq/300 pmol, 3x40 MBq/400 pmol, or 3x60 MBq/600 pmol ¹⁷⁷ Lu-NeoBOMB1. No significant differences in tumor growth delay and median survival were found between treatment groups. This finding was unexpected because preliminary dosimetry calculations using a linear-quadratic model predicted a difference in the probability of tumor control between treatment groups (probability of tumor control: 0%, 75% and 100%, for animals receiving 3×30 MBq/300 pmol, 3×40 MBq/400 pmol and 3×60 MBq/600 pmol, respectively). Without wishing to be bound by theory, it is expected that the dose required to treat the patient will be much lower than expected from previous dosimetric calculations, resulting in lower toxicity of the radioactively labeled NeoBOMB1.

Preferably, the radiolabeled GRPR antagonist carries ¹⁷⁷ Lu.

In specific embodiments of the above methods, the cancer is selected from prostate cancer, breast cancer, small cell lung cancer, colon cancer, gastrointestinal stromal tumors, gastrinomas, renal cell carcinomas, gastroenteropancreatic neuroendocrine tumors, esophageal squamous cell carcinomas, neuroblastomas, head squamous cell carcinomas, and neck, and ovarian, endometrial, and pancreatic tumors displaying neoplasia-associated vasculature that are GRPR positive. In one embodiment, the cancer is prostate cancer or breast cancer.

In one embodiment, the composition to be used is a pharmaceutical composition as described in the previous section.

The invention also relates to a method for treating or preventing cancer, comprising administering to a subject suffering from cancer an effective amount of a composition containing a radioactively labeled GRPR antagonist administered to said subject in a therapeutically effective amount of 2,000 to 10,000 MBq.

Provided herein is a method of treating or preventing cancer, comprising administering to a subject suffering from cancer an effective amount of a composition comprising a radioactively labeled GRPR antagonist as described herein. In some aspects, the cancer is prostate cancer or breast cancer.

In some aspects, the use of a composition comprising a radiolabeled GRPR antagonist in a subject with cancer may inhibit, delay and/or reduce tumor growth in the subject. In some aspects, tumor growth is delayed by at least 50%, 60%, 70%, or 80% compared to an untreated control subject. In some aspects, tumor growth is delayed by at least 80% compared to an untreated control subject. In some aspects, tumor growth is delayed by at least 50%, 60%, 70%, or 80% of predicted tumor growth without treatment. In some aspects, tumor growth is delayed by at least 80% compared to predicted tumor growth without treatment. One of ordinary skill in the art will appreciate that predictions of tumor growth rates can be made based on epidemiological data, reports in the medical literature and other knowledge in the art, tumor type and tumor size measurements, and so on.

In some aspects, the use of a composition containing a radioactively labeled GRPR antagonist in a subject suffering from cancer may increase the length of survival of the subject. In some aspects, an increase in survival is noted compared to an untreated control subject. In some aspects, an increase in survival is noted compared to the predicted duration of survival of the subject without treatment. In some aspects, the duration of survival is increased at least 3-fold, 4-fold, or 5-fold compared to an untreated control subject. In some aspects, the duration of survival is increased by at least 4 times compared with an untreated control subject. In some aspects, the duration of survival is increased by at least 3 times, 4 times, or 5 times compared to the predicted duration of survival in a subject without treatment. In some aspects, the duration of survival is increased by at least 4 times compared to the predicted duration of survival of the subject without treatment. In some aspects, the duration of survival is increased by at least one week, two weeks, one month, two months, three months, six months, one year, two years, or three years compared to an untreated control subject. In some aspects, the duration of survival is increased by at least one month, two months or three months compared to an untreated control subject. In some aspects, the duration of survival is increased by at least one week, two weeks, one month, two months, three months, six months, one year, two years, or three years compared to the predicted duration of survival of the subject without treatment. In some aspects, the duration of survival is increased by at least one month, two months, or three months compared to the predicted duration of survival of the subject without treatment.

In some aspects, the amount of radiolabeled GRPR antagonist administered is less than the amount predicted to give the subject a 100% chance of tumor control.

In some aspects, the amount of the radiolabeled GRPR antagonist is less than the predicted amount to provide the subject with at least a 75% chance of tumor control. In some aspects, the amount of the radiolabeled GRPR antagonist is less than the predicted amount to give the subject a 50% chance of tumor control. In some aspects, the amount of the radiolabeled GRPR antagonist is less than the predicted amount to give the subject a 25% chance of tumor control. In some aspects, the amount of the radiolabeled GRPR antagonist is less than the predicted amount to give the subject a 10% chance of tumor control. In some aspects, the amount of the radiolabeled GRPR antagonist is no more than 25%, 30%, 40%, 50%, 60%, 70%, or 75% of the predicted amount to give the subject a 100% chance of tumor control. In some aspects, the amount of the radiolabeled GRPR antagonist is no more than 50%, 60%, 70%, 75%, 80%, or 85% of the predicted amount to give the subject at least a 75% chance of tumor control. In some aspects, the amount of the radiolabeled GRPR antagonist is no more than 60%, 65%, 70%, 75%, 80%, 85%, or 90% of the predicted amount to give the subject at least a 50% chance of tumor control. In some aspects, the amount of radiolabeled GRPR antagonist administered is a predictable amount that provides the subject with a probability of tumor control of less than 25%, 20%, 15%, 10%, or 5%. In some aspects, the amount of radioactively labeled GRPR antagonist administered is a predictable amount that provides a subject with a 0% chance of tumor control.

Examples

Example 1: Composition screening for NeoBOMB1 adhesion reduction using ⁶⁸ Ga-NeoBOMB1

During the development of the formulation kit, we realized that the peptide has a particular tendency to adhere to glass and plastic surfaces.

This phenomenon is called non-specific binding (NSB). Peptides often exhibit more pronounced NSB problems than small molecules, especially uncharged peptides can be strongly adsorbed to plastics. The reasons can be different: physical/chemical properties, van der Waals interactions, ionic interactions.

An organic solvent can increase solubility and prevent adsorption. Ethanol, for example, can be used in radiopharmaceutical injections to increase the solubility of highly lipophilic indicators or reduce adsorption to vials, membrane filters, and injection syringes. We rejected ethanol because it is incompatible with lyophilization.

Human serum albumin (HSA) is also used in a number of protein formulations as a stabilizer to prevent surface adsorption, but this excipient is unsuitable due to its thermal instability. Another possible approach has been the use of surfactants (eg, polysorbate 20, polysorbate 80, Pluronic F-68, sorbitan trioleate).

We have focused our attention on the study of non-ionic surfactants, since ionic surfactants can interfere with the introduction of the ⁶⁸ Ga label.

Non-ionic surfactants such as Collifor HS 15, Collifor K188, Tween 20, Tween 80, polyvinylpyrrolidone K10 are commercially available as solubilizing auxiliaries in oral and injectable formulations. The table below shows the initial tests that were performed with various surface active agents.

Materials and methods:

NeoBomb1 labeling was based on a previously published approach with the set of Castaldi et al. (Castaldi E, Muzio V, D'Angeli L, Fugazza L. 68GaDOTATATE lyophilized ready to use kit for PET imaging in pancreatic cancer murine model, J Nucl Med 2014; 55(suppl 1):1926).

Various surfactants were screened and the % adhesion of the resulting aqueous solution was determined by calibration measurement of the dose of total radioactivity remaining in the vial after complete removal of the radiolabeled solution. The difference, expressed as a percentage, between total radioactivity measured before and after sample withdrawal is directly correlated with peptide adhesion to the packaging system. The results are summarized in Table 1.

Table 1 - Surfactants - influence on adhesion

NeoBOMB1 (mcg) Solubilizing agent (mcg) Adhesion (%) 50 No surface active agent 21.4 50 PEG 300 (500 mcg) 15.0 50 Ethanol 80% 7.8 50 DMSO 15% 10.4 50 Acetonitrile 80% 7.9 50 PEG 400 (500 mcg) 15.1 50 PVP K10 (500 mcg) 15.6 50 Albumin (500 mcg) 7.9 50 Collifor P188 (1500 mcg) 7.9 50 Hydroxypropyl-β-cyclodextrin (5000 mcg) 13.5 50 PEG 4000 (500 mcg) 12.8 50 Twin 20 (500 mcg) 6.1 50 Kollifor HS 15 (500 mcg) 6.2

The best results in terms of peptide adhesion were obtained with Collifor HS 15 and Tween 20. These two excipients were further tested to determine the final amount in the kit. The results obtained were good in terms of radiochemical purity and peptide adhesion.

Table 2 - Comparison of Twin 20 and Kollifor HS 15

NeoBOMB1 (mcg) Surfactant (mg) ⁶⁸ GaNeoBOMB1 HPLC
(%) Adhesion (%) 50 Twin 20 (1.5 mg) 94.3 5.2 50 Twin 20 (0.5 mg) 94.7 6.0 50 Twin 20 (0.1 mg) 95.3 7.6 50 Kollifor HS 15 (2 mg) 92.8 5.9 50 Kollifor HS 15 (1.5 mg) 93.4 4.9 50 Kollifor HS 15 (1.0 mg) 94.3 4.6 50 Kollifor HS 15 (0.5 mg) 94.9 4.2 50 Kollifor HS 15 (0.25 mg) 95.1 6.0 50 Kollifor HS 15 (0.1 mg) 96.1 8.3 50 Kollifor HS 15 (0.05 mg) 95.7 9.0

We focused on Collifor HS 15 because polysorbates (Tween 20) can undergo auto-oxidation, cleavage at ethylene oxide subunits, and hydrolysis of fatty acid ester bonds caused by the presence of oxygen, metal ions, peroxides, or elevated temperature.

Example 2: Preclinical Studies of Therapeutic Efficacy of ¹⁷⁷ Lu-NeoBOMB1

This application discloses exemplary non-limiting examples of preclinical trials of therapeutic efficacy of ¹⁷⁷ Lu-NeoBOMB1, including treatment of xenograft animals based on the well-known GRPR-expressing prostate cancer cell line PC-3 with 3 different doses of ¹⁷⁷ Lu-NeoBOMB1. In addition, in a small group of non-tumor carrier animals, the effects of ¹⁷⁷ Lu-NeoBOMB1 treatment on the kidneys and pancreas were studied by post-treatment histopathological analysis.

Materials and methods

Introduction of a radioactive label

NeoBOMB1 (Advanced Accelerator Applications) (WO2014052471) was diluted in ultra-pure water, and the concentration and chemical purity were controlled using a proprietary titration method developed (Breeman WA, de Zanger RM, Chan HS, de Blois E. Alternative method to determine the specific activity of 177Lu by HPLC Curr Radiopharm 2015; 8:119–122). A radioactive substance (100 MBq/nmol ¹⁷⁷ Lu) was added to a vial containing all the necessary excipients, eg buffer, antioxidants and peptide, including a surfactant (Collifor HS 15) to prevent sticking of the peptide. High performance liquid chromatography was performed in a gradient of methanol and 0.1% trifluoroacetic acid to determine the radiochemical purity. Radioactive metal incorporation measured by flash thin layer chromatography on silica gel as previously described (de Blois E, Chan HS, Konijnenberg M, de Zanger R, Breeman WA. Effectiveness of quenchers to reduce radiolysis of (111)In- or (177)Lu -labelled methionine-containing regulatory peptides Maintaining radiochemical purity as measured by HPLC Curr Top Med Chem 2012; 12:2677-2685) was >67% and >90% in SPECT/CT and in efficacy and toxicity studies , respectively.

Animal model, efficacy and toxicity

All animal studies were carried out in accordance with the requirements of the Committee for the Humane Treatment of Animals of the Erasmus Medical Center and in accordance with accepted guidelines.

Balb c nu/nu male mice were subcutaneously inoculated into the right shoulder with 200 μl of 4×10 ⁶ PC-3 cells (American Type Culture Collection) in inoculation medium (in 1/3 Matrigel high concentration (Corning) + 2/3 balanced saline Hanks' solution (Thermofisher Scientific)). Four weeks after tumor cell inoculation, upon reaching an average tumor size of 543±177 mm ³ , animals were divided into four groups: control group (n=10) and treatment group 1-3 (n=15 per group). To determine the effectiveness of ¹⁷⁷ Lu-NeoBOMB1, animals received either 3 injections of the solvent (control group) or 3x30 MBq/300 pmol ¹⁷⁷ Lu-NeoBOMB1 (group 1), 3x40 MBq/400 pmol ¹⁷⁷ Lu-NeoBOMB1 (group 2) or 3×60 MBq/600 pmol ¹⁷⁷ Lu-NeoBOMB1 (Group 3) under isoflurane/O ₂ anesthesia. Injections were carried out intravenously and with an interval of 1 week.

To determine the effect of treatment on pancreatic and renal tissues, non-tumor-bearing male Balb c nu/nu mice received the same treatment as the animals included in the efficacy study. At 2 different time points after the last therapeutic injection (12 weeks and 24 weeks), animals were sacrificed and pancreatic and renal tissues were collected for pathological analysis.

In both studies, animal body weight and/or tumor size was measured twice a week. When the tumor size was ≥2000 mm ³ or the decrease in body weight of the animals was ≥20% within 48 hours, the animals were excluded from the study. In the efficacy study, animals were controlled until the maximum allowable age of 230 days was reached.

SPECT/CT

To quantify tumor uptake, SPECT/CT imaging was performed in an additional group of animals with PC-3 xenografts (n = 2 per group). When the tumor size was 477±57 mm ³ , the same amounts of peptides were administered to the animals as to the animals included in the efficacy and toxicity studies. 4 hours and 24 hours after the 1st therapeutic injection, and 4 hours after the 2nd and 3rd therapeutic injections, a whole body scan was performed by SPECT/CT on a hybrid SPECT/CT scanner (VECTor5, MILabs, Utrecht, The Netherlands) . SPECT analysis was performed in 30 minutes with 40 substrate positions using a 2.0 mm point collimator, with a reported spatial resolution of 0.85 mm (Ivashchenko O, van der Have F, Goorden MC, Ramakers RM, Beekman FJ. Ultra-high-sensitivity submillimeter mouse SPECT J Nucl Med 2015;56:470-475). SPECT images were reconstructed using 113 and 208 keV photopeak windows, with a background window on both sides of the photopeak with a width of 20% of the corresponding photopeak, and the corresponding SR-OSEM reconstruction method (Vaissier PE, Beekman FJ, Goorden MC. Similarity-regulation of OS -EM for accelerated SPECT reconstruction Phys Med Biol 2016;61:4300-4315), 0.8 mm ³ voxel size, and CT data were recorded. A subsequent reconstruction of a 3D Gaussian filter (1 mm FWHM) was applied. CT was performed with the following settings: 0.24 mA, 50 kV, full angle scan, 1 position. CT was reconstructed at 100 μm ³ .

Pathological analysis

Pancreas and kidney tissue collected for pathological analysis was fixed with formalin and embedded in paraffin. Hematoxylin and eosin staining was performed on 4 µm tissue sections using the Ventana Symphony™ H&E protocol (Ventana) to determine differences in tissue structure between the 4 treatment groups. A total of 4 tissue sections, separated by 50 μM from each other, were evaluated for each organ. Sections stained with hematoxylin and eosin were evaluated by experienced pathologists.

dosimetry

RADAR realistic mouse model (Keenan MA, Stabin MG, Segars WP, Fernald MJ. RADAR realistic animal model series for dose assessment. J Nucl Med. 2010; 51:471-476) weighing 25 g, and data from previously published studies biodistribution and pharmacokinetics (Dalm SU, Bakker IL, de Blois E, et al. 68Ga/177Lu-NeoBOMB1, a Novel Radiolabeled GRPR Antagonist for Theranostic Use in Oncology. J Nucl Med. 2017; 58:293-299) were used to calculate doses for tumor, pancreas and kidneys when animals were injected with 3×30 MBq/300 pmol, 4×40 MBq/400 pmol or 3×60 MBq/600 pmol ¹⁷⁷ Lu-NeoBOMB1. Biodistribution data from our previously published paper (Dalm SU, Bakker IL, de Blois E, et al. 68Ga/177Lu-NeoBOMB1, a Novel Radiolabeled GRPR Antagonist for Theranostic Use in Oncology. J Nucl Med. 2017; 58:293-299 ) were fitted to exponential curves to determine the time-activity curve in the tumor and organs. Time-integrated activities for ¹⁷⁷ Lu were obtained by integrating these exponential curves stacked with the ¹⁷⁷ Lu decay curve (T _1/2 = 6.647 days). Absorbed doses per administered activity were obtained by multiplying by S-values for organs according to Keenan et al. (Keenan MA, Stabin MG, Segars WP, Fernald MJ. RADAR realistic animal model series for dose assessment. J Nucl Med. 2010; 51:471-476) or using S-values of spherical nodes (Stabin MG, Konijnenberg MW. Re -evaluation of absorbed fractions for photons and electrons in spheres of various sizes J Nucl Med 2000; 41:149-160) for tumor 340 mg.

Tumor dosimetry was used to predict therapeutic outcome using the probability of tumor control (TCP) based on a linear quadratic (LQ) model (Konijnenberg MW, Breeman WA, de Blois E, et al. Therapeutic application of CCK2R-targeting PP-F11: influence of particle range, activity and peptide amount EJNMMI Res. 2014; 4:47).

With N _clonogens , the number of clonogenic (stem) cells in a tumor and S(D,T) of surviving cell fractions is a function of absorbed dose D and time T. The LQ model indicates survival as a function of absorbed dose for tumor growth with twice the time Td:

,

,

where α is the sensitivity of the tumor to radiation, α/β is the ratio between direct (α) and indirect (β) radiation sensitivity, and G is a time factor expressing the development of indirect damage upon dose delivery depending on the half-life of the effective substance and the half-life after sublethal injury. Tumor doubling time was determined by fitting the exponential growth function to tumor volume over time in the control group. Radiation sensitivity parameters for PC-3 tumors were derived from LDR and HDR brachytherapy survival data and α/β = 4.1 (2.5 - 5.7) Gy (Carlson DJ, Stewart RD, Li XA, Jennings K, Wang JZ, Guerrero M. Comparison of in vitro and in vivo alpha/beta ratios for prostate cancer Phys Med Biol 2004;49:4477-4491. The half-life after sublethal injury for PC-3 tumors was 6.6 (5.3 - 8.0) hours (Carlson DJ, Stewart RD, Li XA, Jennings K, Wang JZ, Guerrero M. Comparison of in vitro and in vivo alpha/beta ratios for prostate cancer. Phys Med Biol. 2004; 49:4477-4491), but the value was conservatively fixed at a lower level of 1 hour (Joiner M, Kogel Avd. Basic clinical radiobiology. 4th ed. London: Hodder Arnold ; 2009). The TCP model was used to select which activity to apply that results in growth stunting (TCP = 0%), partial response (TCP >75%), and complete response (TCP = 100%) only. The density of clonogenic cells in PC-3 xenografts was 10 ⁶ cells/cm ³ .

Tumor volume analysis

Tumor doubling time was determined by fitting the exponential growth function to tumor volume over time in the control group. In the treatment groups, the interval with exponential decrease in tumor volume was adjusted to the onset of re-growth after the time of the lowest point. Growth curves were extrapolated beyond the censoring points for mice with too large tumors (>2000 mm ³ ) to determine the average growth statistics. The tumor growth delay time was determined individually by comparing the time required to reach the maximum tumor size of 2000 mm ³ with the average time found in the control group.

Statistics

PRISM software (version 5.01, GraphPad software) was used for statistical analyses. P values >0.05 were considered statistically significant. The difference in tumor volume growth and latency for the 4 groups was established with a one-way ANOVA test with multiple comparative Bonferroni tests. The curves were fitted by least squares with Pearson's R ² test to quantify their suitability.

results

SPECT/CT

At most time points, the mean uptake of radioactivity as measured by SPECT/CT was highest in Group 3, followed by Group 2 and Group 1. However, differences between groups were not significant. Figure 1A shows the scan of one animal of each group obtained at 4 hours and 24 hours after the 1st injection, and 4 hours after the 2nd and 3rd injections. Quantitative tumor uptake is depicted in Figure 1B.

Treatment efficacy ¹⁷⁷ Lu-NeoBOMB1

The ¹⁷⁷ Lu-NeoBOMB1 therapy proved to be effective. In animals in the control group, tumors reached a size of 2000 mm ³ within 20.3 ± 5.9 days, while this size was achieved in 97 ± 59 days, 103 ± 66 days and 95 ± 26 days for group 1, group 2 and group 3, respectively (Figure 2A). In addition, two animals from group 1 and one animal from group 2 did not show any tumors as a result of a complete response. However, there was no significant difference in tumor growth delay time between the treatment groups, while the difference with the control group was very significant (P < 0.0001).

Consistent with the above, animals in the treatment groups had significantly better survival compared to the control group (P<0.001) (FIG. 2B). The median survival was 19 days, 82 days, 89 days and 99 days for the control group, group 1, group 2 and group 3, respectively.

Five animals (n = 3 from group 2 and n = 2 from group 3) were excluded from the study due to the following reasons: 1 animal died after the 1st injection, 1 animal had a very small tumor at the beginning of therapy, which disappeared within several days, 1 animal had a body weight loss of more than 10% within 48 hours, and 1 animal had fluid accumulation in the abdominal cavity. There was no indication that any of the events mentioned were related to treatment.

Toxicity to the kidneys and pancreas

The animals included in the toxicity study did not show a critical reduction in body weight during the entire observation period (Figure 3). The body weight of the animals increased in the first weeks and remained relatively stable over time. One animal in the control group (ID: B) and one animal in group 1 (ID: 869) showed a decrease in body weight, but it was less than 10% within 48 hours.

Histopathological analyzes of the pancreas showed no tissue damage or other abnormalities (Figure 4). With respect to the kidneys (FIG. 5), small areas of lymphocytic infiltration were observed in the kidneys at 12 weeks and 24 weeks after the last therapeutic injection. It was noted in the kidneys of animals in the control group and in the experimental groups, indicating that this phenomenon was not associated with treatment. Twenty-four weeks after treatment, atrophy and fibrosis were observed in the kidneys of only one animal that received the lowest therapeutic dose (ID: 864), and these phenomena are unlikely to be related to treatment. In the kidneys of both animals from group 3, which were sacrificed 24 weeks after the start of treatment, there was a mild chronic inflammatory response.

dosimetry

The dose of radioactivity for the tumor, pancreas and kidneys was evaluated after treatment with 3x30 MBq/300 pmol, 3x40 MBq/400 pmol or 3x60 MBq/600 pmol ¹⁷⁷ Lu-NeoBOMB1 (see Table 3 below). For this, it was assumed that tumor and organ uptake was similar after each injection.

Table 3 Calculated doses for tumor, pancreas and kidney when animals were treated with 3x30 MBq/300 pmol, 3x40 MBq/400 pmol, or 3x60 MBq/600 pmol ¹⁷⁷ Lu-NeoBOMB1*.

Cumulative absorbed dose (Gy) Administered dose 3×30 MBq/ 300 pmol 3×40 MBq/ 400 pmol 3×60 MBq/ 600 pmol Injection 1 ⁱ 2 ⁱ 3 ⁱ 1 ⁱ 2 ⁱ 3 ⁱ 1 ⁱ 2 ⁱ 3 ⁱ Tumor 17 34 68 23 46 91 34 68 137 Bud 1.7 3.4 5.1 2.3 4.5 6.8 3.4 6.8 10 Pancreas 7.9 16 32 eleven 21 42 16 32 64

Claims (32)

1. Directed to the gastrin-releasing peptide receptor (GRPR) pharmaceutical composition containing a radioactively labeled GRPR antagonist having the following formula: MS-S-P, where M is a radioactive metal, and C is a chelator that binds M; S is an optional spacer covalently linked between the C and N-terminus of P; P is a peptide GRP receptor antagonist with the general formula: Xaa1-Xaa2-Xaa3-Xaa4-Xaa5-Xaa6-Xaa7-Z; Xaa1 is the D-isomer of Phe; Xaa2 is Gln; Xaa3 is Trp; Xaa4 is Ala; Xaa5 is Val; Xaa6 is Gly; Xaa7 is His; Z is -NH-alkyl or Z is

, where X is NH and R1 and R2 are the same or different and are selected from proton or optionally substituted alkyl; And a surfactant containing polyethylene glycol-15-hydroxystearate. 2. The pharmaceutical composition according to claim 1, wherein S is a p-aminobenzylamine diglycolic acid residue. 3. Pharmaceutical composition according to claim 1 or 2, where P is DPhe-Gln-Trp-Ala-Val-Gly-His-NH-CH(CH ₂ -CH(CH ₃ ) ₂ ) ₂ . 4. A pharmaceutical composition according to any one of the preceding claims, wherein the GRPR antagonist is NeoBOMB1 of formula (I):

where M is a radioactive metal, preferably M is selected from ¹⁷⁷ Lu, ⁶⁸ Ga. 5. A pharmaceutical composition according to any one of the preceding claims, wherein the radioactively labeled GRPR antagonist is present at a concentration providing a volumetric radioactivity of at least 100 MBq/ml, preferably 250 to 500 MBq/ml. 6. A pharmaceutical composition according to any one of the preceding claims, wherein the surfactant is present at a concentration of at least 5 µg/ml, preferably at least 25 µg/ml, more preferably 50 to 1000 µg/ml. 7. A pharmaceutical composition according to any one of the preceding claims, wherein the radioactively labeled GRPR antagonist carries ¹⁷⁷ Lu, ⁶⁸ Ga, or ¹¹¹ In. 8. Pharmaceutical composition according to any one of the preceding paragraphs, which is an aqueous solution. 9. Pharmaceutical composition according to any one of the preceding paragraphs, which is a solution for infusion. 10. The use of a pharmaceutical composition according to any one of claims 1 to 9 for the treatment or prevention of cancer. 11. Use of a pharmaceutical composition according to any one of claims 1 to 9 for in vivo imaging, preferably PET or SPECT imaging. 12. A method of treating or preventing cancer in a subject in need thereof, comprising administering to said subject a therapeutically effective amount of a composition according to any one of claims 1-9. 13. A method for in vivo imaging of tumors in a subject in need thereof, in particular for detecting GRPR positive tumors, comprising administering to said subject an effective amount of a composition according to any one of claims 1 to 9 and detecting a signal obtained from the decay of a radioisotope present in said compound, thereby allowing the detection of GRPR-positive tumors.

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