# US 8,771,956 B2 · Modulation of human mouse double minute 2 homolog activity

> Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt

- Number: US 8,771,956 B2
- Document ID: US8771956B2
- Status: Granted
- Kind: B2
- Application number: 13/680,263
- Filed: 2012-11-19
- Published: 2013-05-30
- Granted: 2014-07-08
- Earliest claimed: 2003-06-27
- Assignees: Ryogen LLC
- Applicants: Ryogen LLC
- Inventors: James Ryan
- Web page: https://patentgrep.com/patent/US8771956B2

## CPC

- A61K 31/7088: Compounds having three or more nucleosides or nucleotides
- C12N 9/48: acting on peptide bonds (3.4)
- C12Q 1/6876: Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
- C12Q 1/6883: for diseases caused by alterations of genetic material
- C12Y 304/16005: Carboxypeptidase C (3.4.16.5), i.e. carboxypeptidase Y
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- C12Q: MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS (immunoassay G01N33/53); COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Y: ENZYMES

## Abstract

The invention is directed to isolated genomic polynucleotide fragments that encode human carboxypeptidase M and human mouse double minute 2 homolog, vectors and hosts containing these fragments and fragments hybridizing to noncoding regions as well as antisense oligonucleotides to these fragments. The invention is further directed to methods of using these fragments to obtain human carboxypeptidase M and human mouse double minute 2 homolog and to diagnose, treat, prevent and/or ameliorate a pathological disorder.

## Claims

### Claim 1

A method for modulating levels of human mouse double minute 2 homolog activity in a subject in need thereof comprising administering to said subject an amount of an isolated genomic nucleic acid molecule, said nucleic acid molecule selected from the group consisting of:
(a) a nucleic acid molecule consisting of a nucleic acid sequence which has at least 99% identity to the nucleic acid molecule of SEQ ID NO:4 that encodes a polypeptide that has human mouse double minute 2 homolog activity and
(b) a fragment of the nucleic acid molecule of (a), said fragment comprising at least nucleotides 41738-9502 of SEQ ID NO:4 and which encodes a polypeptide having human mouse double minute 2 homolog activity
effective to modulate said human mouse double minute 2 homolog levels.

### Claim 2

A method for modulating levels of human mouse double minute 2 homolog activity in a subject in need thereof comprising administering to said subject an amount of an isolated nucleic acid molecule 20-51039 contiguous nucleotides in length consisting of a reverse or forward strand of a region of SEQ ID NO:4, wherein said region is selected from the group consisting of a 5′-non coding region between nucleotides 51039-41739 of SEQ ID NO:4; a 3′-non-coding region between nucleotides 9503-1 of SEQ ID NO:4; a contiguous intron-exon region between nucleotides 41738-9502 of SEQ ID NO:4, wherein a sequence segment between nucleotides 41738-9502 of SEQ ID NO:4 encodes human mouse double minute 2 homolog depicted in SEQ ID NO:2; a contiguous exon-intron region between nucleotides 41738-9502 of SEQ ID NO:4, wherein a sequence segment between nucleotides 41738-9502 of SEQ ID NO:4 encodes human mouse double minute 2 homolog depicted in SEQ ID NO:2; an intron depicted in nucleotides 36385-40645, 36309-33127, 32994-29616, 29564-25577, 25507-25384, 25287-21169, 21006-14110, 13953-13267, and/or 13188-10665; a region comprising a dinucleotide of the following group: 41739-41738, 40645-40646, 36309-36310, 36384-36385, 32994-32995, 33126-33127, 29564-29565, 29615-29616, 25507-25508, 25287-25288, 25383-25384, 25576-25577, 21006-21007, 21168-21169, 14109-14110, 13953-13954, 13266-13267, 13188-13189, 10664-10665 and/or 9504-9503; a transcription binding site selected from the group consisting of
BINDING SITES
huMDM2, location in SEQ ID NO: 4
AP1_C:
36-46, 2876-2886;
AP4_Q5:
7944-7980;
AP4_Q6:
7943-59, 8924-8940, 9294-9310;
ARNT_01:
1682-1706, 2193-2217, 9201-9225;
BRN2_01:
1040-1058, 7803-7821;
CAAT_01:
3292-3306;
CDPCR3HD_01:
6522-6540;
CEBPB_01:
1424-1438, 3917-3931, 4178-4192, 4787-4801, 6855-6869;
CREL_01:
5630-5642;
DELTAEF1_01:
83-95, 6328-6340;
FREAC7_01:
2757-2773, 5154-5170, 5823-5839;
GATA1_04:
4846-4858, 7017-7029;
GATA1_05:
8464-8476;
GATA2_02:
6045-6057, 6073-6085, 6142-6154;
GATA2_03:
2489-2501, 3323-3335, 3384-3396, 7393-7405:
GATA3_02:
3264-3276, 6870-6882:
GATA3_03:
40-52, 5729-5741, 6529-6541, 6874-6886, 7041-7053, 7589-7601;
GATA_C: 7
349-7361, 8188-8200;
HFH2_01:
1743-1759, 7995-8011;
HFH3_01:
502-518, 1739-1755, 4160-4176, 9402-9418, 9418-9434;
HFH8_01:
8184-8200;
IK2_01:
951-963, 3588-3600;
MZF1_01:
1202-1210, 1447-1455, 4997-4005, 5424-5432;
NF1_Q6:
1480-1500, 8166-8182;
NFAT_Q6:
4190-4208, 6009-6027;
NKX25_01:
741-755, 1648-1662, 1885-1899, 1984-1998, 3609-3623, 4928-4942,
5060-5074, 5889-5903, 8850-8864, 9190-9204;
NKX25_02:
2584-2599, 2970-2984, 4644-4658, 5179-5193, 6482-6496;
NMYC_01:
2560-2572;
RORA1_01:
220-238, 2638-2656;
S8_01:
4644-4656, 4842-4854, 4845-4857, 5200-5212, 5371-5383,
5735-5747, 6482-6494, 6541-6553, 6544-6556, 6772-6784, 7270-7292, 7273-7285;
SOX5_01:
1355-1371, 1430-1446, 3094-3110, 3155-3171, 4669-4685,
4692-4708, 4789-4805;
SRY_02:
4164-4180, 5665-5681;
TATA_01:
1261-1277, 2574-2590, 2723-2739, 2733-2749, 2770-2786,
4199-4215, 4206-4222;
TATA_C:
5900-5916, 7456-7472, 7702-7718, 7917-7933; and
XFD2_01:
7702-7218, 7917-7933;
a transcription binding site selected from the group consisting of
BINDING SITES
huMDM2, location in SEQ ID NO: 4
AP1_C:
12109-12119, 12695-12705, 22600-22610, 24166-24176, 31311-31321,
35234-35244, 39184-39194;
AP1_Q2:
11952-11962, 12068-12078, 14798-14808, 21748-21758, 22613-22623,
23676-23686, 26562-26572, 30046-30056;
AP1_Q4:
12695-12705, 31311-31321, 35234-35244, 36295-36305, 38784-38794,
39188-39198;
AP4_Q6:
31635-31651;
BRN2_01:
13448-13466, 14764-14782, 28094-28112, 40027-40045;
CAAT_01:
11288-11302, 15054-15068;
CDPCR3HD_01:
11286-11304, 13284-13302, 20846-20864, 29344-29362;
CEBPB_01:
29241-29255;
CREL_01:
36091-36103, 38873-38885;
DELTAEF1_01:
18083-18095, 20385-20397, 26955-26967;
FREAC7_01:
11982-11998, 15187-15202, 16523-16539, 16529-16545, 16587-16603,
16604-16620, 16676-16642, 16633-16649, 16644-16660, 16650-16666, 16657-16673, 16673-
16689, 16762-16778, 21332-21348, 25689-25700, 26529-26545, 27767-27783, 29495-29511;
GATA1_02:
10916-10928, 15775-15789, 18162-18174, 26088-26100, 32518-32530;
GATA1_03:
28012-28024;
GATA1_04:
11153-11165, 11630-11642, 13778-13790, 17439-17451, 19300-19312,
21606-21618, 22743-22755, 23747-23759, 25806-25818, 26529-26541, 29424-29436, 30455-
30467, 32761-32778, 33352-33364, 33960-33972, 36101-36113, 40007-40019;
GATA1_05:
11590-11602, 26550-26562, 36737-36749;
GATA1_06:
18772-18784, 23054-23066, 35568-35580, 37855-37867;
GATA2_02:
20755-20767, 30830-30842, 34755-34767, 36285-36297, 39143-39155,
39641-39653, 40586-40598;
GATA2_03:
13535-13547, 22711-22723, 23161-23173, 25028-25040, 27237-27249,
36277-36289;
GATA3_02:
11558-11570, 16470-16482, 17225-17237, 19619-19631, 22156-22168,
22443-22455, 24713-24725, 27619-27631, 32716-32728, 34124-34136, 34163-34175, 36832-
36844, 38403-38415;
GATA3_03:
10869-10881, 11515-11527, 13845-13857, 17221-17233, 18952-18964,
20050-20062, 40171-40183;
GATA_C:
15848-15860, 18899-18911, 23640-23652, 29072-29084, 30881-30893,
33198-33210, 37472-37484, 38621-38633;
GFI1_01:
35469-35481, 35492-35504;
HFH2_01:
15939-15955, 24636-24652, 25866-25882, 32171-32187, 35372-35388,
39457-35473;
HFH3_01:
13340-13356, 19218-19234, 21328-21344, 21336-21352, 21344-21360,
28062-28078, 32125-32141;
HFH8_01:
14133-14149, 22578-22584;
HNF3B_01:
13150-13166, 16505-16521, 25264-25280, 29443-29459, 37654-37670;
IK2_01:
11547-11559, 17144-17156, 18961-18973, 23883-23895, 27617-27629,
28908-28920, 29241-29253, 30752-30764, 34768-34780;
LYF1_01:
12319-12331, 19191-19203, 37226-37238, 39430-39442;
MAX_01:
22974-22986, 33339-33351;
MZF1_01:
26105-26113, 35187-35195;
NF1_Q6:
12048-12064, 33334-33354;
NFAT_Q6:
13295-13313, 14157-14175, 14311-14329, 14414-14432, 18269-18287.
19326-19344, 20801-20819, 21177-21195, 22537-22555, 23861-23879, 25392-25410, 25879-
25897, 27524-27542, 30636-30654, 30718-30736, 31525-31543, 33655-33673, 34726-34744,
34917-34535, 34990-35008, 35979-35997, 36479-36493, 36577-36595, 37154-37172, 40224-
40242, 40365-40383;
NKX25_01:
12041-12055, 12340-12354, 12471-12485, 12742-12756, 12877-12891,
13849-13863, 18995-19009, 21440-21454, 21883-21897, 28426-28440, 30964-30978, 32033-
32047, 32265-32279;
NKX25_02:
10998-11012, 12711-12725, 14131-14145, 14726-14740, 16024-16038;
NMYC_01:
18753-18765, 18754-18766, 23076-23088, 30534-30546, 34400-34412;
RORA1_01:
13134-13152, 22966-22984, 24934-24952, 33341-33359, 34760-34778;
S8_01:
11000-11012, 11977-11989, 12048-12060, 12051-12063, 13747-13759,
13923-13935, 13926-13938, 14676-14688, 14679-14691, 16026-16038, 16313-16325, 16316-
16328, 17515-17527, 20756-20768, 20759-20771, 23154-23166, 23157-23169, 25198-25210,
25201-25213, 26651-26663, 27508-27520, 27511-27523, 29450-29462, 29478-28490, 29775-
29787, 29778-29790, 29813-29825, 29816-29828, 31329-31341, 31677-31689, 31680-31692,
31732-31744, 31735-31747, 36137-36149, 36140-36152, 36812-36824, 36815-36827, 37413-
37425, 38679-38691, 39474-39486, 39477-39489;
SOX5_01:
27397-27413, 27572-27588, 28100-28116, 29230-29246, 29439-29455,
30690-30706, 31595-31611, 33871-33887, 34113-34129, 34624-34640, 37668-37684, 38582-
38598, 39124-39140, 40410-40426;
SRY_02:
20016-20032, 22410-22426, 27329-27345, 29162-29178, 29499-29515,
30646-30662, 31503-31519, 35928-35944, 37324-37340;
TATA_01:
32722-32738, 32729-32745, 32807-32823, 33825-33841, 34120-34136,
35433-35449, 36593-36609;
TATA_C:
11015-11031, 11817-11833, 13635-13651, 14930-14946;
TCF11_01:
18543-18549, 22574-22580, 31281-31297, 31489-31505, 38754-38770;
USF_01:
23075-23087, 32577-32589;
VMYB_02:
11526-11538, 17384-17396, 18400-18412, 19549-19561, 22188-22200,
40486-40508 and
XFD2_01:
16620-16636. 18153-18169, 22102-22118, 23141-23157.
And a transcription binding site selected from the group consisting of
BINDING SITES
huMDM2, 1
location in SEQ ID NO: 4
AP1_C:
44584-44594, 49069-49079:
AP1_Q2:
42174-42184, 45217-45227, 48422-48422, 50447-50457;
AP1_Q4:
42702-42712, 50806-50816;
AP4_Q6:
42117-42133, 42118-42134, 42244-42260, 45432-45448; 45433-45449,
46609-46625;
BRN2_01:
42310-42328, 44022-44040, 47514-47532, 48900-48918, 48967-48985;
CAAT_01:
44866-44880;
CDPCR3HD_01:
45671-45689, 49219-49237;
CREL_01:
42437-42449, 49797-49809;
FREAC7_01:
47026-47042, 47292-47308, 47658-47674;
GATA1_02:
43482-43494, 48926-48938, 49284-49296;
GATA1_03:
47371-47383;
GATA1_04:
43054-43066, 43162-43162, 43967-43979, 45464-45476, 45916-45928,
47763-47775;
GATA1_05:
49319-49331, 49459-49471;
GATA1_06:
47590-47602;
GATA2_02:
42660-42672, 43475-43487;
GATA2_03:
43714-43726, 50948-50960;
GATA3_02:
49155-49167, 49844-49856;
GATA3_03:
42202-42214, 44810-44822, 48438-48450, 49136-49148, 49337-49349,
49869-49881;
GATA_C:
44011-44023, 45256-45268, 45823-45835, 47915-47927, 49201-49213,
49573-49585;
GFI1_01:
46606-46618, 47063-47075;
HFH3_01:
47030-47046, 47284-47300, 47288-47304;
IK2_01:
45275-45287;
LYF1_01:
44564-44576, 46991-47003, 49567-49579;
MAX_01:
43234-43246, 48726-48738;
MZF1_01:
41772-41780, 42290-42298, 42295-42303, 44507-44515, 45105-45113,
45203-45211, 49948-49956, 50774-50782;
NF1_Q6:
50209-50229;
NFAT_Q6:
42061-42079, 44418-44436, 46399-46417, 47974-47992, 49267-49285,
49964-49982, 50392-50410;
NKX25-01:
42394-42408, 43507-43521, 46115-46129;
RORA1_01:
45073-45091, 48718-48736;
S8_01:
43552-43564, 45214-45226, 47160-47172, 48419-48431, 49295-49307,
50379-50391;
SOX5_01:
43716-43732, 46351-46367, 47156-47172, 47774-47790, 47868-47884,
47974-47990, 48915-48931, 50323-50339;
TATA_01:
45588-45604, 47625-47641, 48026-48042, 48659-48675, 49056-49072,
49079-49095, 49152-49168;
TCF11_01:
49115-49131;
VMYB_02:
42010-42022, 42279-42291, 44651-44663; and
XFD2_01:
42870-42886, 42910-42926.